Annual/Termination Reports:

[02/17/2025]

Report Information

Participants

Participants:
In attendance were: Jennifer Barfield (Colorado), Kenneth Bondioli (Louisiana), Celina Checura (South Carolina), Brad Daigneault (Florida), Anna Denicol (California), Jingyue (Ellie) Duan (New York), Zongliang (Carl) Jiang (Florida), Kiho Lee (Missouri), Ying Li (Utah), Charles Looney (Arkansas), Lacey Luense (Texas), Joao Gabriel Nascimento Moraes (Oklahoma), Irina Polejaeva (Utah), Matt Wheeler (Illinois), Ken White (Utah), Curt Youngs (Iowa).

In person attendance (15):
Dr. Jennifer Barfield, Dr. Kenneth Bondioli, Dr. Celina Checura, Dr. Brad Daigneault, Dr. Anna Denicol, Dr. Zongliang (Karl) Jiang, Dr. Kiho Lee, Dr. Ying Li, Dr. Charles Looney, Dr. Lacey Luense, Dr. Joao Gabriel Nascimento Moraes, Dr. Irina Polejaeva, Dr. Matt Wheeler, Dr. Ken White, Dr. Curt Youngs

Zoom attendance (1):
Dr. Jingyue (Ellie) Duan

Brief Summary of Minutes

Brief Summary of Minutes of 2024 Annual Meeting:

Jan 18th, 2025

Worthington Renaissance Fort Worth Hotel, Fort Worth, Texas

Conference room: Post Oak, Upper Level

Zoom link: https://cornell.zoom.us/j/93445516406?pwd=MjV5UzRBbURWbm9DdkdRcGpIY1dBUT09

9:00 AM – 9:15 AM (15 min)

9:00: Dr. Ellie Duan called the meeting to ordered and welcomed the participants.

9:02: All participants introduced themselves in the room and over the Zoom.

9:04: Dr. Ellie Duan presented the agenda for the meeting.

9:05: Dr. Ellie Duan proposed a Motion to approve minutes from 2023 meeting, Dr. Matt Wheeler seconded.  

9:06-09:12: Dr. Matt Wheeler provided a historical commentary on the history of the W171 multistate group and how he and Dr. Curt Youngs had been members for over 30 years. The group was originally started as a Western project, but eventually opened up to the entirety of the country. Discussion continued regarding previous joint publications and the desire to continue to synchronize publication and projects with the current W5171 group.

9:13: Dr. Curt Youngs inquires about including edits for last year’s report submission. Dr. Ken White confirmed that the meeting report could be revised and edited.  Dr. Youngs motioned to approve the minutes with editorial edits. Dr. Celina Checura seconded.

 9:15 AM – 12:00 PM

Station Reports (10-15 min per station) –Station Reports were discussed in alphabetical order.

 Arkansas – Dr. Charles Looney

• Focusing on sex semen and finding the optimal timing of insemination. Working with ST Genetics and other companies.
  
  
  
  • Over four million doses of sexed semen, with an 80% AI rate, and observed no differences in timing between conventional and sexed semen.
  
  
  
  


• Visualizing embryos with Dr. Ken Bondioli (Lousiana).
  
  
  
  • This is an ongoing and productive collaboration.
  
  
  • Identifying non-invasive visualization of ploidy/chromosomal count
  
  
  • Raman spectrometry does not appear to damage embryos
  
  
  • Additional spectral microscopy with NASA
  
  
  
  

 California – Dr. Anna Denicol

• Embryonic stem cell project, with derivation of cultures from early and full blastocysts.
  
  
  
  • Cultures have undergone spontaneous differentiation, formed teratomas, and have performed transcriptomics and determined the developmental competence/primed pluripotency. Trying to develop in less primed state so they can eventually develop into gametes.
  
  
  
  


• Early ovarian follicle project utilizing 3D cultures with biodegradeable flexible matrices with peptide cross-linkers that degrade as follicle grows).


• USDA grant investigating roles of FSH in preantral follicle development.
  
  
  
  • Optimizing conditions for in vivo study
  
  
  • Have developed custom made biopsy needles for OPUs
  
  
  • New study knocking-out FSHb in sheep has resulted in four pregnancies with 80% editing rate.
  
  
  
  


• Overall discussion amongst members about electroporation techniques, methodology, and success.

 Colorado – Dr. Jennifer Barfield

• Using multi-parameter approach to in vitro produced embryo selection for bovine embryo transfer
  
  
  
  • Determining oocyte grade and subsequently tracking embryo development with time lapse live imaging (MIRI). Performing biopsies on d6 or d7 followed by transcriptional analysis of embryos. Collaboration with Todd Hansen at CSU.
  
  
  • Observe a lot of reverse cleavage.
  
  
  • Transcripts of interests identified in biopsies and embryos. Patterns appear to be different based on oocyte grade. Embryo transfers planned for 2025.
  
  
  
  


• Bacterial growth in follicular fluid from beef and dairy cattle. Does not appear pathogenic, but rather contamination.
  
  
  
  • Washing embryos and looking at subsequent microbial contamination.
  
  
  
  


• Additional data from Dr. Dawit Tesfaye will be presented at IETS.


• Karl Jiang discusses collaboration regarding hybrid crosses and aneuploidy in bison.

 Connecticut – not present

 Florida – Drs. Brad Daigenault and Karl Jiang

Daigenault

• Sperm physiology project
  
  
  
  • High resolution respirometry for sperm function utilizing the Seahorse and Oroborus to look at mitochondrial energenetics and oxygen consumption
  
  
  • Sperm consume more oxygen over time and after 24hrs have moved to a more energetic state.
  
  
  • Respiration capacity is reduced in low fertility sires and is moderately correlated to fertility
  
  
  
  


• Capacitation of stallion sperm
  
  
  
  • Working on IVF methods for horses
  
  
  • Determining optimum conditions for sperm prior to IVF by holding sperm at room temperature or cooling and looking at capacitation and motility.
  
  
  • Highest IVF success rate from frozen semen
  
  
  
  


• Recreational pharmaceuticals effects sperm function
  
  
  
  • Altered methylome in human sperm
  
  
  
  


• Embryo project looking at PPARgamma expression and function in preimplantation embryos (USDA-NIFA funded last year).
  
  
  
  • Altered diet and determining if there is an increased embryo developmental competence and pregnancy rate.
  
  
  • Introduction of nanoparticles in embryos
  
  
  
  

 Jiang

• Embryo project focused on building embryos from stem cell cultures.
  
  
  
  • Derivation of bovine extraembryonic endoderm cells (bXENs)
  
  
  • Derivation of bovine naïve embryonic stem cells (bESCs)
  
  
  • d16 elongated embryos and d40 healthy chimeric fetuses from bESCs
  
  
  
  


• Mettl7a improves bovine IVF embryo competence by attenuating oxidative stress


• Ongoing discussion amongst participants about embryo culture, additives, media composition, and success rates. Additional conversation about methods for culturing gonads and chimeras.

 Illinois – Dr. Matt Wheeler

• Embryo transfers in Tanzania
  
  
  
  • Described ongoing work to establish embryo transfer in Tanzania. Must overcome many challenges including bulls from local herders, age of females, and logistical issues.
  
  
  
  


• Beef donors with endocyte infected fescue seed


• Transgenic animals


• Holographic microscopy allows for multiplex optical metabolic imaging (collaboration with engineering school).
  
  
  
  • Embryo metabolism on mouse embryos, label free
  
  
  • Utilizes multi-photon laser to capture sequential imaging of different metabolites.
  
  
  • Scan through embryos for metabolites using machine learning/AI.
  
  
  
  


• Significant discussion amongst members about holographic microscopy and potential uses. Multiple investigators interested in learning more and potential collaborations.

 Missouri – Dr. Kiho Lee

• Announced that Dr. Randy Prather has retired and he is currently transitioning into the position of the head of the swine center.


• Reprogramming factors, lineage specification, and epigenetic regulation in swine
  
  
  
  • Prdm family important for blastocyst formation, may have role in ZGA.
  
  
  • Transcriptional analysis through qPCR and bulk RNAseq have identified stage and lineage gene expression differences
  
  
  
  


• Investigating novel off-targeting gene editing events
  
  
  
  • Determining ways to tell the difference between genetics and off-targeting
  
  
  
  


• Improving genome editing with Cas9 variants to improve editing.
  
  
  
  • Selected variants based on human and mouse efficiency.
  
  
  
  


• Ahmed Balboula is working on improved quality of in vitro produced embryos


• Group discussion on genome editing, injections, and efficiency of different Cas9 variants and methodologies.

 12:00 End of morning session

 12:00 – 13:25 – Lunch break

 13:25 Start of afternoon session

 13:25 – 15:30 – Continuing station reports

Iowa – Dr. Curt Youngs

• Ewe vaginal microbiota
  
  
  
  • Large sample size (30 ewes sampled per week), pregnant vs pregnant
  
  
  • Shift in microbiota when progesterone takes over
  
  
  • Microbiota shifted from beginning of breeding season, looking for biomarker
  
  
  
  


• Sheep in vitro embryo production
  
  
  
  • Collaboration with group in Peru, looking at differences in maturation buffer, particularly when working at high elevation (and higher CO2%)
  
  
  • Transfers have been completed
  
  
  
  


• Synchronization of ovulation – postpartum dairy cattle
  
  
  
  • Trying to move away from as many injections for synchronization (less labor, less hormone)
  
  
  • Comparing 1 vs 2 PGF injections. 1 injection did not change pregnancy rate compared to double ov sync
  
  
  • No difference in total pregnancy loss
  
  
  
  

 Louisiana – Dr. Ken Bondioli

• In vitro oocyte maturation
  
  
  
  • Utilizing c-type natriuretic peptide in pre maturation culture
  
  
  • Addition of L-carnitine to IVM medium and looking at COC expansion and membrane potential ROS/ATP, molecular profiling with Raman spec
  
  
  • COC expansion during in vitro maturation, conventional is best for 6 hours, but carnitine and CNP treatments improved over 28 hours
  
  
  • CNP improved mitochondrial membrane potential, ROS, but not ATP
  
  
  • Raman spectrometry is able to see shift in molecular fingerprint with CNP
  
  
  • CNP treatment improved mitochondrial membrane potential in bovine oocytes, L-carnitine increased ROS, and Etomoxir prevented molecular alterations in CNP treated oocytes
  
  
  
  


• Vigorous Injection Methods for ICSI
  
  
  
  • Increased cleavage (82%) and blast rate in fresh oocytes (75%)
  
  
  • More cells in vitrified embryos, including ICM and TE
  
  
  • Have produced 3/10 transfers maintained to term, 2 fetuses had severe dystocia and did not survive, one was delivered after induction
  
  
  
  

 Mississippi – not present

 Montana – not present

 Nebraska – not present

 New York – Dr. Ellie Duan

• Functional genomics and epigenomics in early embryogenesis
  
  
  
  • MOF/H4K16ac is present in bovine early embryo and ZGA
  
  
  • siRNA knock-down of MOF, followed by molecular/genomic analysis. MOF is maternally expressed, having challenges knocking down, however microinjection seems to be working
  
  
  
  

• Characterize molecular difference during sex-specific embryogenesis (post-doc presenting – Peter Farin Trainee Award)
  
  
  
  • Sex specific differences in embryogenesis. RNAseq from half of a blast (other half for determine embryo sex)
  
  
  • Faster development in male embryos
  
  
  • Dynamics of gene expression and transposable elements in bovine early embryos
  
  
  • Data mining from other epigenetic marks that are already published in multiple species
  
  
  
  

• Impact of heat stress on early embryo development
  
  
  
  • Cows exposed to heat stress chamber, identifying key regulators of heat stress in dairy cow livers and on milk genes
  
  
  • Comparative genomics and transcriptomics in development and aging
  
  
  • Sex specific aging – epigenetic mechanisms and evolution (IISAGE) – large NSF funded project (role is to integrate multi-omics data)
  
  
  • Identifying pluripotency regulators in ESCs across species
  
  
  • Comparing stem cells across species to compare/find pluripotency regulators, comparing different states in different stages
  
  
  • Developed pipeline for other ES cell work to determine type/fit of cell based on transcriptomics
  
  
  
  

 Oklahoma – Dr. Joao Gabriel Nascimento Moraes

• Nascimento Moraes was presenting at an IETS pre-conference symposium and pre-recorded a video of his update.


• Due to technical difficulties with the AV system in the conference room, the video was unable to play in the room and was later viewed by attendees


• Role of small non-coding RNAs in embryo development
  
  
  
  • Changes in transcriptome between ovoid and tubular stages of development
  
  
  • Differentially expressed miRNA and tRFs
  
  
  
  


• Establishment of protocols and conditions for culturing ESC from Bos taurus (female) vs Bos bison (male) hybrid embryos
  
  
  
  • Established four porcine fetal fibroblasts cell lines and will knock-out candidate genes via gene editing
  
  
  
  

 South Carolina – Dr. Celina Checura

• Research primarily focuses on undergraduate projects (undergrads presenting two posters at IETS)


• Feeding fescue seeds to sheep and determining the metabolic potential of oocytes.


• Photo maturation project
  
  
  
  • No effect when completed after ZGA
  
  
  • May be effecting granulosa cells and not oocyte?
  
  
  
  


• Semen analysis effects on IVF looking at ROS and DNA fragmentation


• Brad Daigenault suggests that an investigator at Florida is interested in developing collaborations for feeding fescue to horses.


• Group discussion about use of Seahorse for mitochondrial studies with potential collaborative potential.

 Texas – Dr. Lacey Luense

• Epigenetic regulation of paternal histone epigenome and embryo
  
  
  
  • Studies in mouse demonstrate that H3K27ac is present at genes that are paternally expressed
  
  
  • Identified potential binding motifs where H3K27ac is enriched, including ZGA pioneer factors Nr5a2
  
  
  
  


• Role of paternal sperm histones on embryo development and pregnancy loss in cattle
  
  
  
  • Looking at sperm histone profiles in high and low fertility sires. Low fertility sires have previously been linked to pregnancy loss
  
  
  • Characterization of histone post-translational modifications in bull testes and preimplantation embryos
  
  
  
  

 Utah – Dr. Ying Liu

• Retrospective analysis of sheep generated by somatic cell nuclear transfer
  
  
  
  • Identified factors that affect or don’t affect SCNT efficiency
  
  
  • Link to pregnancy loss
  
  
  • Age of oocyte donor does not impact in vitro maturation rate, pregnancy and full-term development, SCNT efficiency
  
  
  • Earlier activation of SCNT embryos was correlated w/lower early pregnancy loss rate, higher full term rate
  
  
  
  


• Cytokine-supplementation of maturation medium enhances cytoplasmic and nuclear maturation in bovine oocytes
  
  
  
  • FLI supplementation in bovine oocyte maturation medium improved nuclear progression through meiosis I and proper meiotic spindle formation
  
  
  
  

 15:30 Discussion about new collaborations

• Ken White describes how he has heard multiple conversations about research stations with shared expertise. These include conversations during the station reports and those during breaks and lunch. He encourages the development of three or four groups to focus on research questions. Not every necessarily needs to participate, but we should focus on strengths.


• Matt Wheeler suggests the sharing of materials to advance progress. Materials could include oocytes, recipients, semen, etc.


• Discussion from multiple investigators about techniques that they are currently troubleshooting and possible insight.


• Matt Wheeler suggest using Whatsapp to have a group chat to facilitate communication throughout the year.

 15:50 Meeting business

 Nomination for Chair and Secretary of the W5171 2025-2026 Annual Meeting

• Dr. Ellie Duan nominated Dr. Ying Li for secretary of the 2026 meeting. Dr. Brad Daigneault seconded.


• Dr. Ellie Duan nominated Dr. Lacey Luense for chair of the 2026 meeting. Dr. Curt Youngs seconded.

 2026 Meeting Venue, Date and Time – Member discussion

• Possible dates for the 2025 annual meeting will be near IETS time, which is 2026 1.16-1.20 in Panama City, Panana.


• Due to IETS being held at an international location, alternative dates and locations were discussed, including having it before or after SSR, having it at an alternative location, or having it at a US based location immediately before IETS.


• Participants voted to hold the meeting at a US location that would be en route to Panama, possibly Houston or Florida. (8 ayes – 1 vote per station).


• Once the schedule of activities for IETS has been released, there will be a Doodle Pool to decide the time and location for the 2026 annual meeting.


• Duan discussed payment options for the room and AV rental.

 16:10 Dr. Curt Youngs motioned to adjourn the 2025 meeting. Seconded by Dr. Matt Wheeler.

 

 

 

Accomplishments

<p><strong>ACCOMPLISHMENTS</strong></p><br /> <p><strong><span style="text-decoration: underline;">Objective 1</span></strong><strong>- Understand the biology of gamete development, fertilization and embryogenesis including the underlying cellular and molecular mechanisms.</strong></p><br /> <ol><br /> <li>Improvements in sexing technologies may offer comparable pregnancy outcomes for both conventional and sexed semen, regardless of insemination time.</li><br /> <li>Optical imaging offers a potential non-invasive and accurate avenue for assessing embryo viability</li><br /> <li>Bovine embryonic stem cell (bESC) lines were established from early (eBL) and full (BL) blastocyst-stage embryos to determine the efficiency of bESC derivation from an earlier embryonic stage and compare the characteristics of the resulting lines. Transcriptome and functional analyses indicated that eBL- and BL-derived lines were in primed pluripotency. Indistinguishable bESC lines can be readily derived from eBL and BL, widening the pool of embryos available for bESC establishment.</li><br /> <li>Culture of primary stage preantral follicles in PEG hydrogels promoted growth compared to controls; BOCs did not maintain identity in the PEG hydrogels. Collectively, we demonstrate that PEG hydrogels can be a potential culture system for early preantral follicles pending refinements, which could include addition of ESC-derived ovarian somatic cells using the protocol described here.</li><br /> <li>Investigating the impact of genetic and pharmacological approaches to modulate NRF2-mediated oxidative stress response in bovine preimplantation embryos will provide a unique opportunity to enhance survival and mitigate the long-term impact of preimplantation period oxidative damage on fetal development and offspring health. Moreover, the study will identify genes and genome regions which can be targets for future interventions to enhance viability of embryos from assisted reproductive technologies.</li><br /> <li>Investigated maternal age dependent changes in granulosa cells and follicular fluid miRNA profile and clear molecular alterations can be seen in granulosa cells and EVs derived from follicular fluid EVs, which can explain age associated infertility problems in human.</li><br /> <li>While there are potentially pathogenic bacteria in the follicular fluid, they are effectively removed by the in vitro embryo production process with or without trypsin washing of the embryos.</li><br /> <li>Demonstrated the ability to identify expression levels of transcripts of interest regarding embryo competence in embryo biopsies and are analyzing these data relative to oocyte quality and developmental kinetics.</li><br /> <li>Studied the mechanism of ploidy correct in pre-implantation embryos.</li><br /> <li>Single-cell transcriptomic analysis of bovine blastocysts produced in vivo and in vitro in conventional and reduced nutrient conditions reveals the effect of culture environments on embryo developmental potential.</li><br /> <li>We describe a molecule (METTL7A) that acts in the pre-implantation period to attenuate oxidative stress that enhances embryo development to the blastocyst stage and subsequent pregnancy in cattle.</li><br /> <li>Using a high-resolution ribosome profiling approach, we analyzed the transcriptome, as well as monosome- and polysome-bound RNAs of mouse oocytes and embryos, defining an unprecedented extent of translational landscapes during this rapid developmental phase. We showed that a eukaryotic initiation factor Eif1ad3, which is exclusively translated in the 2-Cell embryo, is required for ribosome biogenesis post embryonic genome activation.</li><br /> <li>Single-cell transcriptomes analysis of bovine peri-implantation embryo development at day 12, 14, 16 and 18 post fertilization, when most pregnancy failure occurs.</li><br /> <li>We profile the transcriptome of two widely used stem cell-based trophoblast models, compare to trophoblasts from peri-implantation human embryos, and validate several genes in peri-implantation human embryos with immunofluorescence</li><br /> <li>Optimized Oroboros instrumentation for high resolution respirometry measurements of frozen-thawed bull sperm.</li><br /> <li>Determined that postejaculatory mammalian sperm are susceptible to epigenetic modifications.</li><br /> <li>Characterized responses of postejaculatory bovine sperm to environmental stressors and recreational pharmaceuticals.</li><br /> <li>Optimized sperm conditions and defined temporal aspects of stallion sperm capacitation conducive to equine in vitro fertilization.</li><br /> <li>Identified strategies to improve the shelf-life of cooled stallion sperm for artificial insemination.</li><br /> <li>Defined alternative strategies for producing multiple bovine embryos from a single 4-cell embryo by blastomere complementation. Such strategies also advance trophectoderm models of early embryo development.</li><br /> <li>Characterized and determined functions of peroxisome proliferator-activated receptor gamma in bovine embryo development.</li><br /> <li>Results show that heat-stress (HS) can compromise the recruitment of ovarian follicles early in a new follicular wave.</li><br /> <li>Using the commercial protocol, high cleavage rate and blastocyst rates were achieved. Large numbers of embryos will be needed to improve the dairy cattle genetics in Africa. The efficiency of the present IVF system will enable that large-scale embryo production.</li><br /> <li>The transcriptome, molecular fingerprint and mitochondrial function of bovine oocytes matured in vitro with a conventional 22h procedure were compared to oocytes matured after a pre maturation culture in the presents of C-type natriuretic peptide by RNA sequencing and Raman spectroscopy. Differences in the transcriptome , molecular fingerprint of lipid droplets, mitochondrial function and fatty acid oxidation which may impact developmental competence of subsequent embryos were characterized.</li><br /> <li>Identifying Cathepsin V as a novel target to improve the quality of in vitro produced embryos.</li><br /> <li>Development and validation of effective protocols for in-house synthesis of heavy sulfur ([34S])-labeled methionine and cystine.</li><br /> <li>Train and develop expertise in microsurgical insertion of jugular catheters in mice for whole-mouse metabolic labeling studies.</li><br /> <li>(with collaborators) Develop and validate protocols for measuring most biologically important sulfur amino acid metabolites by liquid chromatography-mass spectrometry.</li><br /> <li>Quantitation of the flux of the sulfur and all carbon atoms from methionine into downstream metabolites in the methionine cycle, methionine salvage pathway, transsulfuration, and glutathione biosynthesis pathways, as well as several other key pathways.</li><br /> <li>Quantitation of the flux of the sulfur and all carbon atoms from cystine into downstream metabolites in transsulfuration, per / poly-sulfide production, and glutathione biosynthesis pathways, as well as several other key pathways.</li><br /> <li>Quantitative verification of the role of the methionine catabolism pathway in redox homeostasis.</li><br /> <li>Identification of a novel non-canonical non-reductive cystine catabolism pathway that also supports redox homeostasis.</li><br /> <li>Characterize the dynamic of MOF/H4K16ac during preimplantation embryogenesis via qPCR and immunofluorescence staining</li><br /> <li>Established the CUT&amp;Tag-T7 protocol to profile MOF/H4K16ac genomic distribution in low cell number embryo samples</li><br /> <li>Sexed embryos using the PCR method.</li><br /> <li>Generated full-length transcriptomic analysis of male and female blastocysts.</li><br /> <li>Results will be presented as a poster at IETS 2025, the manuscript is under preparation.</li><br /> <li>Compared the transcriptomic profiles of bovine embryos produced from different technologies.</li><br /> <li>Characterized the impact of embryo production technologies on TE dynamics and regulation.</li><br /> <li>Results will be presented as a poster at IETS 2025, the manuscript is under preparation.</li><br /> <li>Generated novel microRNA and tRNA-derived fragment (tRF) data during bovine conceptus elongation.</li><br /> <li>Windows of sensitivity to photobiomodulation at different stages of gamete and embryo development have been found. Photobiomodulation is being used as a tool to understand the role of mitochondria at different developmental stages.</li><br /> <li>Identified that paternal H3K27ac is present at transcription factors associated with embryonic genome activation in mouse preimplantation embryos.</li><br /> <li>Established in vitro embryo production in laboratory.</li><br /> <li>Characterized H3K27ac in bovine testis and preimplantation embryos.</li><br /> <li>FLI supplementation in bovine oocyte maturation medium improved nuclear progression through meiosis I and increased proper meiotic spindle formation, as well as both mitochondrial and cortical granule redistribution.</li><br /> <li>FLI supplantation accelerates SCNT embryo development, and reduces morphokinetic activity during early development, suggesting that FLI supplementation in the maturation medium serves as protection from environmental stress.</li><br /> <li>The retrospective analysis identified parameters, including earlier activation time and commercial serum-free media, for improving pregnancy and full-term development of SCNT embryos in sheep.</li><br /> <li>A general decrease in CPEB1 expression levels occur from GV and MII oocytes to blastocyst stage.</li><br /> <li>Lower concentrations of CoQ10 supplementation do not affect oocyte maturation and quality or embryo development, whereas a higher concentration of CoQ10 increases mitochondrial mass in bovine oocytes and decreases their cleavage rate after PA.</li><br /> </ol><br /> <p><strong><span style="text-decoration: underline;">Objective 2</span> - Refine methods to produce animals by genetic engineering or genome editing for the improvement of livestock production efficiency and development of human biomedical models.</strong></p><br /> <ol><br /> <li>Used lentiviral-mediated, RNA interference (RNAi) to target genes specifically in sheep trophectoderm. By infecting day 9 hatched blastocyst with replication-deficient lentivirus, only the outer trophectoderm cells are infected, effectively limiting the RNAi to the developing placenta.</li><br /> <li>Several ongoing sheep pregnancies using in vivo placenta-specific lentiviral-mediated RNAi methodologies to interrogate the role of placental TTR and DIO2 during the first-half of gestation.</li><br /> <li>Started a project to refine the embryo complementation technology for the one-step generation of genetically engineered large animals.</li><br /> <li>Conducted studies for the separation of mouse, bovine and human sperm for sex selection.</li><br /> <li>Establishment of bovine extraembryonic endoderm cells (XENs) from pre-implantation embryos</li><br /> <li>Through 3D assembly of bXENs with bovine ESCs and TSCs, we developed an improved bovine blastocyst like structure (bovine blastoid) that resembles blastocyst.</li><br /> <li>Demonstrated feasibility of alternative approaches to delivering plasmids in bovine embryos for overexpression studies.</li><br /> <li>Established a model for conducting transgenic studies in bovine embryos</li><br /> <li>Demonstrated that mature bioactive human insulin can be produced in cows milk, which could potentially revolutionize how insulin is administered to diabetic patients.</li><br /> <li>A modified procedure for intracytoplasmic sperm injection (ICSI) was used to fertilize vitrified and non-vitrified bovine oocytes. This procedure yielded a higher proportion of blastocyst compared to a conventional procedure with a greater cell number in the inner cell mass and trophectoderm. It was demonstrated that these ICSI derived embryos can produce pregnancies and live birth upon transfer.</li><br /> <li>Detection of off-targeting events in genome edited pigs.</li><br /> <li>Validated effectiveness of Cas9 variants.</li><br /> <li>Functional validation of AAV8-sgRNA vectors targeting specific genes for disruption of gene function and downstream physiological impacts in hepatocytes of liver-specific Cas9-expressing mice.</li><br /> <li>Established embryonic stem cell lines from Bos taurus (female) vs Bos bison (male) hybrid embryos.</li><br /> <li>Established porcine fetal fibroblast cell lines for use in genome editing experiments.</li><br /> <li>By modifying the mitochondrial bioenergetic status of blastocyst, the cell numbers were enhanced, not only in the cryopreservation group (not statistically significant) but also significantly in the non-cryopreservation group. This indicates that the blastocysts are sensitive to the photobiomodulation treatment and that there is room to improve the <em>in vitro</em> embryo production system through mitochondria enhancement.</li><br /> <li>Retrospective data analysis conducted using data from seven different projects (aimed to produce genetically engineered animals) showed no differences between the projects in pregnancy and full development rates, and cloning efficiency. Additionally, no difference was observed between male and female donor cells used as donor cell lines.</li><br /> </ol>

Publications

<p><strong>Publications</strong></p><br /> <ol><br /> <li>Denicol AC. What we can learn from the bovine embryo and mouse models to enable in vitrogametogenesis in cattle. Reproduction, Fertility and Development 37, RD24142. 2025. doi:10.1071/RD24142</li><br /> <li>Altman MA, Mathews AT, Rabaglino MB, Hovey RC, Denicol AC. Canonical prolactin signaling and global mRNA expression in the skin of Holstein heifers carrying the SLICK1 allele of the prolactin receptor gene. J of Dairy Sci 2025 (ACCEPTED).</li><br /> <li>Candelaria JI, Botigelli RC, Guiltinan C, Shikanov A, Denicol AC. Three-dimensional culture in a bioengineered matrix and somatic cell complementation to improve growth and survival of bovine preantral follicles. bioRxiv [Preprint]. 2024 Aug 7:2024.07.18.604061. doi: 10.1101/2024.07.18.604061.</li><br /> <li>Kennedy VC, Lynch CS, Tanner AR, Winger QA, Gad A, Paul J. Rozance PJ, Anthony RV. Fetal Hypoglycemia Induced by Placental SLC2A3 RNA Interference Alters Fetal Pancreas Development and Transcriptome at Mid-Gestation. Int. J. Mol. Sci. 2024, 25(9), 4780; https://doi.org/10.3390/ijms25094780.</li><br /> <li>Tanner AR, Kennedy VC, Lynch CS, Winger QA, Anthony RV, Rozance PJ. Increasing maternal glucose concentrations is insufficient to restore placental glucose transfer in chorionic somatomammotropin RNA interference pregnancies. Am J Physiol Endocrinol Metab. 2024 Feb 14.</li><br /> <li>Xiao-Kim E, Graham JK, Schountz T, Barfield JP. 2024. Osmotic tolerance and cryopreservation of Jamaican fruit bat (Artibeus Jamaicensis) sperm. Therio Wild, vol 6, 100118.</li><br /> <li>Nico G Menjivar, Jaiden Oropallo, Samuel Gebremedhn, Luca A Souza, Ahmed Gad, Christian M Puttlitz, Dawit Tesfaye. 2024. MicroRNA Nano-Shuttles: Engineering Extracellular Vesicles as a Cutting-Edge Biotechnology Platform for Clinical Use in Therapeutics. Biol Proced Online 26, 14.</li><br /> <li>Joyce K, Gad A, Menjivar NG, Gebremedhn S, Heredia D, Dubeux G, Lopez-Duarte MC, Bittar J, Gonella-Diaza A, Tesfaye D. 2024. Seasonal environmental fluctuations alter the transcriptome dynamics of oocytes and granulosa cells in beef cows. J Ovarian Res;17(1):201.</li><br /> <li>Feugang JM, Gad A, Menjivar NG, Ishak GM, Gebremedhn S, Gastal MO, Dlamini NH, Prochazka R, Gastal EL, Tesfaye D. 2024. Seasonal influence on miRNA expression dynamics of extracellular vesicles in equine follicular fluid. J Anim Sci Biotechnol;15(1):137.</li><br /> <li>Dessie Salilew-Wondim, Michael Hoelker, Eva Held-Hoelker, Franca Rings, Ernst Tholen, Christine Gro&szlig;e-Brinkhaus, Karl Shellander, Carina Blaschka, Urban Besenfelder, Vita Havlicek, Dawit Tesfaye. 2024. Sexual dimorphic miRNA-mediated response of bovine elongated embryos to the maternal microenvironment. PLoS ONE 19(2).</li><br /> <li>Ahmed Gad, Nico G Menjivar, Rachel Felton, Barbara Durrant, Dawit Tesfaye, Elena Ruggeri. 2024. Mapping the follicle-specific regulation of extracellular vesicle-mediated microRNA transport in the southern white rhinoceros (Ceratotherium simum simum). Biology of Reproduction. ioae081.</li><br /> <li>Eva Held-Hoelker, Jessica Kurzella, Dessie Salilew-Wondim, Franca Rings, Dawit Tesfaye, Ernst Tholen, Christine Grosse-Brinkhaus, Michael Hoelker. 2024. L-Carnitine sustainably affects bioenergetic profile of bovine blastocysts and transcriptome profile of elongation-stage embryos. Reproduction Aug 2;168(3):e240004.</li><br /> <li>Jessica Kurzella, Dennis Miskel, Franca Rings, Ernst Tholen, Dawit Tesfaye, Karl Schellander, Dessie Salilew-Wondim, Eva Held-Hoelker, Christine Gro&szlig;e-Brinkhaus, Michael Hoelker. 2024. Mitochondrial bioenergetic profiles of warmed bovine blastocysts are typically altered after cryopreservation by slow freezing and vitrification. Theriogenology 214, 21-32.</li><br /> <li>Jessica Kurzella, Dennis Miskel, Franca Rings, Ernst Tholen, Dawit Tesfaye, Karl Schellander, Dessie Salilew-Wondim, Eva Held-Hoelker, Christine Gro&szlig;e-Brinkhaus, Michael Hoelker: 2024. Mitochondrial bioenergetic profiles of warmed bovine blastocysts are typically altered after cryopreservation by slow freezing and vitrification. Theriogenology 214, 21-32.</li><br /> <li>Zhu J, Lai Y, Cheng M, Dash RC, Guo S, Guo J, Su Y, Wolek A, Issacs B, Liu Z, Li Q, Mishra N, *Garmendia A, *Hadden MK, *Tian XC, *He X, *Tang Y. 2024. Discovery of Small Molecules Against Porcine Reproductive and Respiratory Syndrome Virus Replication by Targeting NendoU Activity. Journal of Virology ; <a href="https://doi.org/10.1128/jvi.02034-24">https://doi.org/10.1128/jvi.02034-24</a></li><br /> <li>Ming H, Scatolin G, Ojeda A, Jiang Z#. Establishment of bovine extraembryonic endoderm cells. bioRxiv [Preprint]. 2024 Dec 18. https://biorxiv.org/cgi/content/short/2024.12.17.628911v1</li><br /> <li>Zhu L, Ming H, Scatolin G, Xiao A, Jiang Z#. METTL7A improves bovine IVF embryo competence by attenuating oxidative stress. bioRxiv [Preprint]. 2024 Dec 18. https://biorxiv.org/cgi/content/short/2024.12.17.628915v1</li><br /> <li>Ming H, Iyyappan R, Kakavand K, Dvoran M, Susor A, Jiang Z#. Spatiotemporal dynamics and selectivity of mRNA translation during mouse pre-implantation development. bioRxiv [Preprint]. 2024 Oct 28:2024.10.28.620693. doi: 10.1101/2024.10.28.620693. PMID: 39553972.</li><br /> <li>Logsdon DM, Ming H, Ezashi T, West RC, Schoolcraft WB, Roberts RM, Jiang Z#, Yuan Y#. Transcriptome comparisons of trophoblasts from regenerative cell models with peri-implantation human embryos&sbquo;&Auml;&dagger;. Biol Reprod. 2024 Nov 11;111(5):1000-1016. doi: 10.1093/biolre/ioae120. PMID: 39109839.</li><br /> <li>Ming H, Zhang M, Rajput S, Logsdon D, Zhu L, Schoolcraft WB, Krisher RL, Jiang Z#, Yuan Y#. In vitro culture alters cell lineage composition and cellular metabolism of bovine blastocyst&sbquo;&Auml;&dagger;. Biol Reprod. 2024 Jul 12;111(1):11-27. doi: 10.1093/biolre/ioae031. PMID: 38408205.</li><br /> <li>Znachorova T, Dudko N, Ming H, Jiang Z, Fulka H. The timing of pronuclear transfer critically affects the developmental competence and quality of embryos. Mol Hum Reprod. 2024 Jun 26;30(7):gaae024. doi: 10.1093/molehr/gaae024. PMID: 38991843.</li><br /> <li>Xu S, Wang N, Zuccaro MV, Gerhardt J, Iyyappan R, Scatolin GN, Jiang Z, Baslan T, Koren A, Egli D. DNA replication in early mammalian embryos is patterned, predisposing lamina-associated regions to fragility. Nat Communications. 2024 Jun 19;15(1):5247. doi: 10.1038/s41467-024-49565-7. PMID: 38898078.</li><br /> <li>Lamacova L, Jansova D, Jiang Z, Dvoran M, Aleshkina D, Iyyappan R, Jindrova A, Fan HY, Jiao Y, Susor A. CPEB3 Maintains Developmental Competence of the Oocyte. Cells. 2024 May 16;13(10):850. doi: 10.3390/cells13100850. PMID: 38786074.</li><br /> <li>Scatolin GN, Ming H, Wang Y, Iyyappan R, Gutierrez-Castillo E, Zhu L, Sagheer M, Song C, Bondioli K, Jiang Z#. Single-cell transcriptional landscapes of bovine peri-implantation development. iScience. 2024 Mar 27;27(4):109605. doi: 10.1016/j.isci.2024.109605. PMID: 38633001.</li><br /> <li>Fendereski M, Ming H, Jiang Z, Guo Y Mouse trophoblast cells have attenuated responses to TNF and IFN and can avoid synergic cytotoxicity of the two cytokines.. Journal of Immunology. 2024 Jan 15. PMID: 38054905.</li><br /> <li>Guertin JE, Losano JDA, Salazar S, Callaham J, Daigneault, BW. Prolonged maintenance of cooled stallion semen by optimization of storage conditions. Journal of Equine Veterinary Sciences. 2024 Nov 19;144:105243. doi: 10.1016/j.jevs.2024.105243. Online ahead of print.</li><br /> <li>McGraw MJ, Bishman JA, Daigneault BW. Blastomere complementation efficiency and pluripotency maintenance following multiple passaging of in vitro derived bovine embryos. Reproduction, Fertility and Development. 2024 Jun:36:RD24018. doi: 10.1071/RD24018.</li><br /> <li>Camila J. Cuellar,Thiago F. Amaral, Paula Rodriguez-Villamil, F. Ongaratto, D. Onan Martinez,Remi Labreque,Jo D.A. Losano,Eliab Estrada-Cortes, Jonathan R. Bostrom, Kyra Martins, D. Owen Rae, Jeremy Block, Quinn A. Hoorn, Bradford W. Daigneault, Jonathan Merriam, Michael Lohuis, Serdal Dikmen, Joao Bittar,Tatiane S. Maia, Daniel F. Carlson, Sabreena Larson, Tad S. Sonstegard, and Peter J. Hansen. Consequences of gene editing of PRLR on thermotolerance, growth, and reproduction in cattle. FASEB Bioadv. 2024 2024 Jun 18;6(8):223-234. doi: 10.1096/fba.2024-00029. eCollection 2024 Aug</li><br /> <li>Cesar A. Rosales-Nieto, Bradford W. Daigneault, Jennifer N. Roberts, Rodrigo Sanchez-Lopez, Barbara Makela, Yong Pu, Richard Ehrhardt, Kristi L Jones, Joao H. Jabur Bittar, Almudena Veiga-Lopez. Birth weight, growth indices, and seminal parameters in male offspring are resilient features to maternal pre-conceptional dietary manipulation in sheep. Domest Anim Endocrinol. 2024 Apr 7:88:106849. doi: 10.1016/j.domaniend.2024.106849.</li><br /> <li>McGraw MJ, Rajput SK, Daigneault BW. Peroxisome proliferator-activated receptor gamma influences developmental competence and early placental formation in bovine embryos. Reproduction. 2024 Jan 31;167(2):e230334. doi: 10.1530/REP-23-0334. Print 2024 Feb 1.</li><br /> <li>Stella, S., Guadagnin, A.R.,Velasco-Acosta, D., Ferreira, C., Rubessa, M., Wheeler, M.B., Luchini, D., Cardoso, F. (2024) Rumen-Protected Methionine Supplementation Alters Lipid Profile of Preimplantation Embryo and Endometrial Tissue of Holstein Cows. Front. Vet. Sci., 17 January 2024. Sec. Animal Nutrition and Metabolism, Volume 10 - 2023 | https://doi.org/10.3389/fvets.2023.1301986</li><br /> <li>Wheeler, M.B., Rabel, R.A. Chanaka, Rubessa, M., and Gabriel Popescu (2024) Invited Paper - Label-Free, High-Throughput Holographic Imaging to Evaluate Mammalian Gametes and Embryos. Biology of Reproduction, 2024, ioae057, https://doi.org/10.1093/biolre/ioae057</li><br /> <li>Monzani, P.S., Sangalli, J.R., Sampaio, R.V., Guemra, S., Zanin, R., Adona, P.R., Berlingieri, M.A., Cunha-Filho, L.F.C, Mora- Ocampo, I.Y., Pirovani, C.P, Meirelles, F.V., Wheeler, M.B., Ohashi, O.M. (2024). Human proinsulin production in the milk of transgenic cattle. Biotechnology Journal, 19, e2300307. https://doi.org/10.1002/biot.202300307</li><br /> <li>Geng Wang, Lianhuang Li, Xiaoxia Liao, Shu Wang, Jennifer Mitchell, R.A. Chanaka Rabel, Shirui Luo, Jindou Shi, Janet E. Sorrells, Rishyashring R. Iyer,</li><br /> <li>Edita Aksamitiene, Carlos A. Renteria, Eric J. Chaney, Derek J. Milner, Matthew B. Wheeler, Martha U. Gillette, Alexander Schwing,Jianxin Chen, Haohua Tu (2024). Supercontinuum intrinsic fluorescence imaging heralds &lsquo;free view&rsquo; of living systems. bioRxiv https://doi.org/10.1101/2024.01.26.577383.</li><br /> <li>Bangert, E. A., C. Allen, A. Chota, G. Msuta, E. Mbisha and M. B. Wheeler (2024). Assessment of Challenges and Recommendations for Success When Implementing Assisted Reproductive Technology Programs in Developing Countries in the Tropics (Tanzania).</li><br /> <li>CABI Animal Science Cases 2024. DOI: 10.1079/animalsciencecases.2024.0012</li><br /> <li>Rabel, R.A.C., Bangert, E.A., Wilson, K., Wheeler, M.B. (2024). The Use of Assisted Reproductive Technologies to Improve Genetic Selection in Cattle. In: Gard&oacute;n, J.C., Satu&eacute; Ambrojo, K. (eds) Assisted Reproductive Technologies in Animals Volume 1. Springer, Cham. <a href="https://doi.org/10.1007/978-3-031-73079-5_8">https://doi.org/10.1007/978-3-031-73079-5_8</a></li><br /> <li>Sun, N.N. Ali, D. Londo&radic;&plusmn;o-V&radic;&deg;squez, C.A. Simintiras, H. Qiao, M.S. Ortega, Y. Agca, M.M. Takahashi, R.M. Rivera, A.M. Kelleher, P. Sutovsky, A.L. Patterson, A.Z. Balboula*. (2024) Increased DNA damage in full-grown oocytes is correlated with diminished autophagy activation. Nature Communications, 1, 15(1): 9463. * Corresponding author.</li><br /> <li>Kiesler ZG, Hunter MI, Balboula AZ, Patterson AL. (2024). Periostin's role in uterine leiomyoma development: a mini review on the potential periostin poses as a pharmacological intervention for uterine leiomyoma. Archives of Gynecology and Obstetrics, 309(5): 1825-1831.</li><br /> <li>Elmetwally MA, Helmy A, Zaabel S, Montaser A, Balboula A, Eladl M, Hamed B, Lashen S, Rezk S, Yaseen A, Sharawy H, Hussien M, Halawa A. (2024). Fructose improves Titanium Dioxide Nanoparticles induced alterations in Developmental Competence of mouse oocytes. BMC Veterinary Research, 3, 20(1): 135.</li><br /> <li>Redel BK, Yoon J, Reese E, An H, Uh K, Chen PR, Prather RS, Lee K. Novel off-Targeting Events Identified after Genome Wide Analysis of CRISPR-Cas Edited Pigs. CRISPR J. 2024 May 21. doi: 10.1089/crispr.2024.0012. Online ahead of print.</li><br /> <li>Sun, P. Sutovsky, A.L. Patterson, A.Z. Balboula AZ*. (2023). Mechanisms of DNA damage response in mammalian oocytes, Advances in Anatomy, Embryology and Cell Biology series. 238:47-68. * Corresponding author.</li><br /> <li>Arner, E.S.J., Schmidt, E.E., 2024. Unresolved questions regarding cellular cysteine sources and their possible relationships to ferroptosis. Advances in cancer research 162, 1-44.</li><br /> <li>Borb&eacute;nyi-Galambos, K., Erd&eacute;lyi, K., Ditr&oacute;i, T., Jur&aacute;nyi, E.P., Sz&aacute;nt&oacute;, N., Szatm&aacute;ri, R., Czikora, &Aacute;., Schmidt, E.E., Garai, D., Cserepes, M., Liszkay, G., T&oacute;th, E., T&oacute;v&aacute;ri, J., Nagy, P., 2025. Realigned Transsulfuration Drives BRAF V600E-Targeted Therapy Resistance in Melanoma. Cell Metabolism In revision.</li><br /> <li>Mart&iacute;-Andr&eacute;s, P., Finamor, I., Torres-Cuevas, I., P&eacute;rez, S., Rius-P&eacute;rez, S., Colino-Lage, H., Guerrero-G&oacute;mez, D., Morato, E., Marina, A., Michalska, P., Le&oacute;n, R., Cheng, Q., Jur&aacute;nyi, E.P., Borb&eacute;nyi-Galambos, K., Mill&aacute;n, I., Nagy, P., Miranda-Vizuete, A., Schmidt, E.E., Mart&iacute;nez-Ruiz, A., Arn&eacute;r, E.S., Sastre, J., 2024. TRP14 is the rate-limiting enzyme for intracellular cystine reduction and regulates proteome cysteinylation. The EMBO Journal 43, 2789-2812.</li><br /> <li>Schmidt, E.E., Miller, C.G., Juranyi, E.P., Austad, S.N., Ditroi, T., Seaford, Z.M., Yoon, S.J., Noyd, R.C., Kang, Y.P., Prigge, J.R., Ciskos, V., Kovari, D., DeNicola, G.M., Nagy, P., 2025. Cystine C-S bond cleavage fuels cysteine production under disulfide reductase deficiency. Natue Chemical Biology In revision.</li><br /> <li>R.Biga*, J. E. Duan*, T E. Young*, J. R. Marks*, A. Bronikowski3*,L. P. Decena3*, E. C. Randolph1*, A. G. Pavuluri*, G. Li*, Y. Fang*, G. S. Wilkinson*, G. Singh#, N. T. Nigrin6#, E. N. Larschan6#, A. J. Lonski#, Nicole C. Riddle1^, and IISAGE Consortium (2024). Hallmarks of Aging: A User's Guide for Comparative Biologists, in press. Ageing Research Reviews</li><br /> <li>Su,Y., Zhao, R., Fang, Y., Li, G., Jin, L., Liu, J., Zhu, J., Mishra, N., Kaback, D., Yee, S., Lei, C., Duan, J.*, Tian, X.*, Tang, T.*&nbsp; (2024) Under revision. Bovine Formative Embryonic Stem Cells Capable of Three-Germ Layer and Extraembryonic Differentiation.</li><br /> <li>Yu X., Harman R., Danev N., Li G., Fang Y., Van de Walle G., Duan J.E*. Under review. Heat stress and recovery induce transcriptomic changes in lactogenic bovine mammary epithelial (MAC-T) cells. 2024 bioRxiv, https://www.biorxiv.org/content/10.1101/2024.05.15.594241v1</li><br /> <li>Aguilera J., Duan J., Lee S., Ray M., Larschan E. The CLAMP GA-binding transcription factor regulates heat stress-induced transcriptional repression by associating with 3D loop anchors. bioRxiv 2023.10.08.561401; doi: https://doi.org/10.1101/2023.10.08.561401</li><br /> <li>Li G, Danev, N, Duan, J*, Van de Walle G. (2024), Comparative transcriptomic analysis of bovine mesenchymal stromal cells reveals tissue-source and species-specific differences. 2024 Jan 12;27(2):108886. doi: 10.1016/j.isci.2024.108886. PMID: 38318381; PMCID: PMC10838956.</li><br /> <li>F.F. B&radic;&part;nmann, L.G.D. Mendon&radic;&szlig;a, I. Sellmer Ramos, R. Fritz, C. Gamarra, D. Duhatschek, R.S.S. de Oliveira, A.L.A. Scanavez, T.S. Belem, M.C. Lucy, J.G.N. Moraes. Sensor-based and Visual Behavioral Profiling of Dry Holstein Cows Presenting Distinct Median Core Body Temperature. doi.org/10.3390/ani14192832. Animals 14, no. 19 (2024): 2832.</li><br /> <li>G.N. Moraes, S. E. Poock, A.C. Ericsson, T. Gull, Caldeira, M.O., and M. C. Lucy. 2024. The microbiome of the pregnant uterus in Holstein dairy heifers and cows assessed by bacterial culture and 16S ribosomal RNA gene sequencing. Front Microbiol. May 15:15:1385497. doi: 10.3389/fmicb.2024.1385497. eCollection 2024.</li><br /> <li>G.N. Moraes, T. Gull, A.C. Ericsson, S. E. Poock, Caldeira, M.O., and M. C. Lucy. 2024. Establishment of the uterine microbiome following artificial insemination in virgin heifers. Front Microbiol. Jun 5:15:1385505. doi: 10.3389/fmicb.2024.1385505. eCollection 2024.</li><br /> <li>Sellmer Ramos, M. O. Caldeira, S. E. Poock, J.G.N. Moraes, M. C. Lucy, and A.L. Patterson. Adenomyosis and fibrosis define the morphological memory of the postpartum uterus of dairy cows previously exposed to metritis. JDS Communications. 2024 Oct 30. doi.org/10.3168/jdsc.2024-0633</li><br /> <li>G. da Silva, L.G. da Silva, L.C.L. Ferreira, J. Mascarello, J.C.B. Silva, J.G.N. Moraes, M.C. Lucy, E. Nogueira. Factors influencing pregnancy per artificial insemination (AI) and embryonic mortality in Nelore females subjected to timed-AI in Brazil. Anim Reprod Sci. 2024 Jun:265:107475. doi: 10.1016/j.anireprosci.2024.107475.</li><br /> <li>C. C. Silva, M. O. Caldeira, J.G.N. Moraes, I. Sellmer Ramos, T. Gull, A. C. Ericsson, S. E. Poock, T. E. Spencer, and M. C. Lucy. 2024. Metritis and the uterine disease microbiome are associated with long-term changes in the endometrium of dairy cows. Biol Reprod. 2024 May 5:ioae67. doi: 10.1093/biolre/ioae067.</li><br /> <li>Ing, N.H., Curley Jr., K.O., Welsh Jr., T.O., Johnson, L., Luense, L.J., Clement, T.M. &ldquo;Anatomy and Physiology of the Male Reproductive System and Potential Targets of Toxicants.&rdquo; In Comprehensive Toxicology, 3^rd Edition, Elsevier Science, 2024, <em>In press</em></li><br /> <li>Dodd, A.R., Luense, L.J. Contribution of the paternal histone epigenome to the preimplantation embryo. 2024 Frontiers in Cell and Developmental Biology, 12, doi:0.3389/fcell.2024.1476312</li><br /> <li>Blocher, R., Liu, Y., Patrick, T., Polejaeva, I.A.* (2024) Cytokine-supplemented maturation medium enhances cytoplasmic and nuclear maturation in bovine oocytes. Animals, 14:1837.</li><br /> <li>Liu, Y*., Stott, R., Regouski, M., Fan, Z., Viotti Perisse, I., Patrick, T., Keim, J., Meng, Q., Polejaeva, I.A.* (2024) A retrospective analysis of sheep generated by somatic cell nuclear transfer. Theriogenology 227:102-111.</li><br /> <li>Leir, S-H., Tkachenko, S., Paranjapye, A., Meckler, F., Van Wettere, A.J., Kerschner, J.L., Kuznetsov,E., Schacht, M., Giiurkar, P., Regouski M., Viotti Perisse, I., Marriott C.M., Liu, Y., Bunderson, I., White, K.L., Polejaeva, I.A., Ann Harris* (2024) Stellate cells are in utero markers of pancreatic disease in cystic fibrosis. Molecular Medicine&nbsp; 30:115.</li><br /> <li>Adams, L., Liu, Y., Polejaeva, I.A.* (2024) Current status of interspecies somatic cell nuclear transfer and meta&sbquo;&Auml;&ecirc;analysis of the effects of phylogenetic distance on embryonic and fetal development. Mammal Review. 54:387-403.</li><br /> </ol><br /> <p><strong>Books, non-refereed book chapters, proceedings, instructional media, theses/dissertations</strong></p><br /> <ol><br /> <li>Acevedo, Carolina. Development of a bison-specific embryo culture system through targeted supplementation of media with stage-specific growth factors, Spring 2024, in fulfillment of MS degree. Advisee of J. Barfield.</li><br /> <li>Gene-environment interactions contributing to early embryo development in cattle. Maura M. McGraw, PhD Thesis, University of Florida.</li><br /> <li>Optimization of sperm conditions for equine reproductive technologies. Jillian, E Guertin, MS Thesis, University of Florida.</li><br /> <li>Comparison of isolation procedures for cryopreserved stallion sperm compatible with equine in vitro fertilization. Lana Katz, University of Florida Undergraduate Scholar Thesis.</li><br /> <li>Brewer, A. &ldquo;A Novel Goat Model to Characterize Male Fertility Following Paternal cannabidiol Exposure&rdquo;. Louisiana State University Masters Thesis. 2024.</li><br /> <li>Gatenby, L. &ldquo;Improving Bovine Intracytoplasmic Sperm Injection.&rdquo; Louisiana State University Ph. D. dissertation. 2024.</li><br /> <li>Girka, E. &ldquo;Metabolic Activity During Bovine Maturation in Vivo and in Vitro&rdquo;. Louisiana State University Ph. D. Dissertation.&nbsp;</li><br /> </ol><br /> <p><strong>Abstracts</strong></p><br /> <ol><br /> <li>Midkiff, Kristen A., Kegley, Elizabeth B., Cheek, Robin A., Galloway, Doug L., Reynolds, Jana L., Harness, Alexis, Looney, Charles R., Littlejohn, Brittni P., Powell, Jeremy G. (2024).&nbsp; Effect of diets containing soybean co-products formulated for developing heifers on reproductive efficiency.&nbsp; Journal of Animal Science.&nbsp; 102 (Supp. 3):356-357.&nbsp; https://doi.org/10.1093/jas/skae234.407.&nbsp; Graduate student (Ph.D.) presented in Oral Abstracts &ndash; Ruminant Nutrition.</li><br /> <li>Efird, Emily. E., Powell, Jeremy G., Looney, Charles R., Perkins, Tommy L. (2024).&nbsp; Evaluation of factors influencing conception rate within a fixed-time artificial insemination protocol using sex-sorted semen.&nbsp; Journal of Animal Science.&nbsp; Vol 102 (Supp. 3):474-475.&nbsp; https://doi.org/10.1093/jas/skae234.539.&nbsp; Graduated student presented poster abstract in ASAS Graduate Student Poster Competition &ndash; MS.</li><br /> <li>Snider, Miriam A., Culp, Timothy C., Hopkins, Carson R., Moss, Nathaniel T., Edwards, J. Lannett, Looney, Charles R., Powell, Jeremy G., Kegley, Elizabeth B., Coffey, Ken P.; Gadberry, M. Shane, Lemley, Caleb. O., Littlejohn, Brittni P. (2024).&nbsp; Influence of melatonin supplementation in pregnant multiparous crossbred beef cows grazing endophyte-infected tall fescue on dam weight, milk yield, and pre-weaning growth of steer calves.&nbsp; Journal of Animal Science.&nbsp; Vol 102(Supp. 3):206-207.&nbsp; https://doi.org/10.1093/jas/skae234.242.&nbsp; Graduate student presented oral abstract &ndash; Growth and Development.</li><br /> <li>Snider, Miriam A., Culp, Timothy C., Hopkins, Carson R., Moss, Nathaniel T., Edwards, J. Lannett, Looney, Charles R., Powell, Jeremy G., Kegley, Elizabeth B., Coffey, Ken P. Gadberry, M. Shane, Lemley, Caleb O., Littlejohn, Brittni P. (2024).&nbsp; Influence of melatonin supplementation on uterine and coccygeal artery hemodynamics of pregnant multiparous crossbred beef cows grazing endophyte-infected tall fescue.&nbsp; Journal of Animal Science.&nbsp; Vol102(Supp.3):284-285.&nbsp; https://doi.org/10.1093/jas/skae234.326.&nbsp; Graduate student presented oral abstract &ndash; Physiology and Endocrinology.</li><br /> <li>Efird, Emily. E., Gilbreath, E., Powell, Jeremy G., Moss, Nathaniel T., Gadberry, M. Shane, Looney, Charles R. (2024).&nbsp; Effect of Semen Type and Fixed-Time Artificial Insemination Protocol on Pregnancy Rates for Beef Cows and Heifers.&nbsp; Applied Reproductive Strategies in Beef Cattle Conference.&nbsp; Graduate student presented oral abstract.</li><br /> <li>Showe, Clayton E., Weber, Wyatt E., Kutz, B. R., Shoulders, Benjamin P. Shoulders, Reynolds, Jana L., Powell, Jeremy P., Looney, Charles R. (2024).&nbsp; Comparison of GnRH Administration Day in the 7 &amp; 7 Synch Versus 8 &amp; 6 Synch Protocol for Fixed-Time Artificial Insemination in Beef Heifers.&nbsp; Journal of Animal Science.&nbsp; 101 (Supp. 1):57-58.&nbsp;</li><br /> <li>Denicol, Anna, 2025 Conference Abstracts. Reproduction, Fertility and Development 37, RDv37n1abs. doi:10.1071/RDv37n1abs</li><br /> <li>Singleton SA, Kennedy VC, Donovan A, Anthony RV, Winger QA. Placenta-Specific RNA Interference of Transthyretin in Sheep. Society for the Study of Reproduction 56th Annual Meeting, Dublin, Ireland.</li><br /> <li>Kennedy VC, Tanner AR, Lynch CS, Winger QA, Rozance PJ, Anthony RV. Impact of Placental SLC2A3 RNA Interference throughout Gestation. Society for Reproductive Investigation 71st Annual Scientific Meeting, Vancouver, BC.</li><br /> <li>Russell V Anthony, Amelia R Tanner, Victoria C Kennedy, Quinton A Winger, Paul J Rozance. 9 Randel Lecture: In Vivo investigation of pregnancy Physiology Journal of Animal Science, Volume 102, Issue Supplement_1, March 2024, Pages 52&ndash;53.</li><br /> <li>Rogers H, Gad A, Cork G, Menjivar N, Schoolcraft WB, Tesfaye D, Yuan Y. Age-related transcriptomic profiling in human granulosa cells reveals mitochondrial insufficiency and microRNA regulation of key ovarian microenvironment dynamics. Fertility and Sterility 122:e168. (Poster Presentation) https://doi.org/10.1016/j.fertnstert.2024.07.587</li><br /> <li>Thompson RE, Gad A, Meyers MA, Menjivar NG, Tesfaye D, Hollinshead FK. Evaluation of extracellular vesicles secreted by mouse oviductal organoids for loading with CRISPR-cas9 ribonucleoproteins. Society for the Study of Reproduction 2024; 57th Annual Meeting. Dublin, Ireland. (Oral Presentation)</li><br /> <li>Thompson RE, Gad A, Meyers MA, Menjivar NG, Tesfaye D, Hollinshead FK. Extracellular Vesicles Secreted by Oviductal Organoids for Delivery of CRISPR-Cas9 Ribonucleoproteins. Rocky Mountain Reproductive Sciences Symposium 2024; 16th Annual Conference: Pages 54-55. Fort Collins, CO, USA. (Poster Presentation)</li><br /> <li>Wittenstein J, Gad A, Menjivar N, Tesfaye D. Strategies to Combating Oxidative Stress Damage During Bovine Embryo Development. Rocky Mountain Reproductive Sciences Symposium 2024; 16th Annual Conference: Page 42. Fort Collins, CO, USA. (Poster Presentation)</li><br /> <li>Ram&iacute;rez GG, Gad A, Menjivar NG, Tesfaye D. Antioxidant-Based Strategies To Mitigate The Impact Of Heat Stress On Bovine Granulosa Cells And Oocytes. Rocky Mountain Reproductive Sciences Symposium 2024; 16th Annual Conference: Page 29. Fort Collins, CO, USA. (Poster Presentation)</li><br /> <li>Rogers H, Gad A, Cork G, Menjivar N, Schoolcraft WB, Tesfaye D, Yuan Y. Age-Related Transcriptomic Signatures in Human Granulosa Cells Emphasize Inflammation, Extracellular Matrix Remodeling, and Angiogenesis. Rocky Mountain Reproductive Sciences Symposium 2024; 16th Annual Conference: Page 17. Fort Collins, CO, USA. (Oral Presentation)</li><br /> <li>Gad A, Menjivar NG, Tesfaye D. The power of extracellular vesicles enhancing animal fertility. CSU Demo Day 2024; Annual Showcase. Fort Collins, CO, USA. (Poster Presentation)</li><br /> <li>Gad A, Menjivar NG, Tesfaye D. Fertility Resilience in the Face of Climate Change: The Role of Extracellular Vesicles. PASS Research Symposium 2024: Page 3-4. Fort Collins, CO, USA. (Oral Presentation)</li><br /> <li>Wittenstein J, Gad A, Menjivar N, Tesfaye D. Enhancing the antioxidant capacity of bovine embryos against oxidative stress using quercetin supplementation. CVMBS Research Day 2024: Page 120. Fort Collins, CO, USA. (Poster Presentation)</li><br /> <li>Rogers H, Gad A, Cork G, Menjivar N, Meyers M, Tesfaye D, Yuan Y. Transcriptomic insights into advanced maternal aged granulosa cells emphasizes inflammation and immune response. CVMBS Research Day 2024: Page 110. Fort Collins, CO, USA. (Poster Presentation).</li><br /> <li>Dunn B, Menjivar NG, Gad A, Thompson RE, Meyers MA, Hollinshead FK, Tesfaye D. Generation of physiologically relevant extracellular vesicles from bovine oviductal organoids. CVMBS Research Day 2024: Page 88. Fort Collins, CO, USA. (Poster Presentation).</li><br /> <li>Ram&iacute;rez GG, Gad A, Menjivar NG, Tesfaye D. Enhancing the antioxidant capacity of granulosa cells to mitigate the impact of heat stress. CVMBS Research Day 2024: Page 46. Fort Collins, CO, USA. (Oral Presentation).</li><br /> <li>Braun H, Gad A, Barfield J. Time-Lapse Assessment of Bovine Embryos from Varying Oocyte Quality. Graduate Student Showcase 2024. Fort Collins, CO, USA. (Abstract / Poster Presentation)</li><br /> <li>Braun H, Gad A, Barfield J. Developmental potential and morphokinetics of in vitro produced bovine embryos generated from oocytes of varying quality. IETS 2025 Annual Conference. Fort Worth, TX, USA. (Abstract for Poster Presentation)</li><br /> <li>Braun H, Guzeloglu A, Bishop J, Georges H, Menjivar N, Marquez M, Hansen T, Barfield J. Identification and evaluation of transcriptomic markers for assessing bovine embryo quality from individual embryos or biopsies. CVBMS 26th Annual Research Day. Fort Collins, CO, USA. (Abstract for Oral Presentation)</li><br /> <li>Bakhtari A, Zhao R, Liu J, Su Y, Bielski K, and *Tian X. 2025. Ploidy correction during pre-implantation development of bovine triploid embryos. Poster at the 51st annual meeting of the International Embryo Technology Society, January 17-21, 2025, Fort Worth, TX, USA (published in Reproduction Fertility and Development, 2025.</li><br /> <li>Zhao R, Bakhari A, Liu J, *Tian X. 2025. Triploid Complementation: A Potential Alternative Approach to its Tetraploid Counterpart. Poster at the 51st annual meeting of the International Embryo Technology Society, January 17-21, 2025, Fort Worth, TX, USA (published in Reproduction Fertility and Development, 2025.</li><br /> <li>Losano JDA, Souders C, Binelli M, Martyniuk C, Daigneault BW. Beyond Motility - Advanced Bioenergetic Assessment of Bull Sperm. National Association of Animal Breeders 2024</li><br /> <li>Losano JDA, Guertin JE, McGraw M, Turner C, Pontes JHF, Fleury P, Maserati M, Callaham J, Bordignon V, Daigneault BW. Functional Capacitation of Stallion Sperm Promotes Interspecies Embryo Chimerism and Equine In Vitro Blastocyst Development from Cryopreserved Sperm. Society for the Study of Reproduction 2024.</li><br /> <li>Mohammad SS, Gauthier M, Martyniuk CJ, Kladde M, Daigneault BW. Postejaculatory Sperm Exposure to Tributyltin Chloride Induces Novel Structural, Genomic and Epigenomic Alterations in Mammalian Sperm. Society for the Study of Reproduction 2024.</li><br /> <li>Venturini M, Losano JDA, Heredia D, L&radic;&ge;pez M, McGraw M, Muhammad S, Guertin J, DiLorenzo N, Daigneault BW, Gonella A. Association between residual feed intake and reproductive-related traits in young bulls. American Society of Animal Sciences 2024.</li><br /> <li>Monzani, P.S., Sangalli, J.R., Sampaio, R.V., Guemra, S., Zanin, R., Adona, P.R., Berlingieri, M.A., Cunha Filho, L F.C., Mora-Ocampo, I.Y., Pirovani, C P., Meirelles, F.V., Ohashi, O. and Wheeler M.B. (2024). Human proinsulin and insulin production in the milk of transgenic cattle. Reproduction, Fertility and Development 36, (2) 231 https://doi.org/10.1071/RDv36n2Ab155</li><br /> <li>Marchioretto, P.V., Rodriguez-Zas, S L., Womack, S.A., Lindsey, B.R., Milner, D.J., Rubessa, M., Wilson, K.C. and Wheeler, M.B. (2024). Effect of dominant follicle removal before ovum pickup in Girolando cattle. Reproduction, Fertility and Development 36, (2) 251-252 https://doi.org/10.1071/RDv36n2Ab193</li><br /> <li>Glassey, J.R., Rabel, R.A.C., Milner, D.J. and Wheeler, M.B. (2024) Strontium enhances in vitro osteogenic differentiation of porcine adipocyte&ndash;derived stem cells. Reproduction, Fertility and Development 36, (2) 265-266 https://doi.org/10.1071/RDv36n2Ab220 &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</li><br /> <li>Bangert, E., Shipley, C., Rabel, R.A.C., Garrett, E., Milner, D. J., Marchioretto, P.V., Spencer, K., Allen, C., and Wheeler, M.B. (2024). The efficiency of an adapted bovine IVF protocol to produce in vitro-derived embryos from oocytes collected via surgical ovum pickup from live white-tailed deer (Odocoileus virginianus) donors under captivity in central Illinois. Reproduction, Fertility and Development 36, (2) 165-166 https://doi.org/10.1071/RDv36n2Ab32</li><br /> <li>Zimmerman, L.A., Bangert, E. A., Rabel, R. A C., Milner, D. J., Marchioretto, P.V., Allen, C.A. and Wheeler, M.B. (2024). The in vitro production of Gyr X Jersey bovine embryos from oocytes collected via ovum pickup for use in the tropics. Reproduction, Fertility and Development 36, (2) 155 https://doi.org/10.1071/RDv36n2Ab12&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</li><br /> <li>Womack, S.A., Bethke, E.B., King, W P., Milner, D.J., Rubessa, M., Marchioretto, P.V., and Wheeler, M.B. (2024). Development of a porcine model for the testing of the RapidVent emergency ventilator for the treatment of COVID-19 infection. Reproduction, Fertility and Development 36, (2) 219 https://doi.org/10.1071/RDv36n2Ab132 &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</li><br /> <li>Wheeler, M.B. (2024). Awardee Talk - The Quest for an Ideal In-Field Embryo Evaluation Tool in Cattle: Opportunities and Challenges. Journal of Animal Science, Volume 102, Issue Supplement_3, September 2024, Pages 287&ndash;288, https://doi.org/10.1093/jas/skae234.329</li><br /> <li>Kim J, Yoon J, Chen J, Lee H, Redel B, Prather RS, Lee K. Enhancing specificity of gene editing outcomes by using Cas9 variants in swine embryos. IETS Jan 18 &ndash; 22. 2025.</li><br /> <li>Yoon J, Mony S, Kiesewetter E, Sullivan R, Kim J, Redel B, Uh K, Prather RS, Lee K. Effects of stem cell factor on the in vitro maturation of porcine oocytes and subsequent development following in vitro fertilization. IETS Jan 18 &ndash; 22. 2025.</li><br /> <li>Montgomery T, Uh K, Prather RS, Lee K. Identifying PRDM family members potentially involved in epigenetic reprogramming after fertilization. IETS Jan 18 &ndash; 22. 2025.</li><br /> <li><span style="text-decoration: underline;"> Li</span> and J. Duan, 2024. Diversity of cow transposable elements and developmental expression in early embryos. Poster presentation at The Allied Genetics Conference (TAGC), Washington DC</li><br /> <li><span style="text-decoration: underline;">Jingzhi Zhang*,</span> Chia-Kuan Wu*, Meihong Shi, Abdallah Abdelhady, Yoke Lee Lee, Soon Hon Cheong&dagger;, Ellie Duan&dagger;. Profiling of H4K16 Acetyltransferase MOF in Bovine Embryogenesis. 2024 Poster presentation at Tri-Reproduction Symposium, Pittsburg, PA</li><br /> <li><span style="text-decoration: underline;"> Li</span>, M. Shi, H. Araujo, A. Lee, J. Zhang, L. Lee, S. H. Cheong, J. E. Duan, 2024 Sex Differences of Bovine Blastocyst Revealed By Transcriptome Profiling and Long-Read Sequencing. To be presented as a poster presentation at Plant &amp; Animal Genome 32, San Diego, CA</li><br /> <li><span style="text-decoration: underline;"> Li</span> and Duan, J. E Comparative analysis of gene expression and transposable elements&nbsp;dynamics&nbsp;in bovine early embryos across different conditions. To be presented as a poster presentation at the 51st annual meeting of the International Embryo Technology Society, Fort Worth, TX</li><br /> <li><span style="text-decoration: underline;"> Shi</span>, G.Li, H. Araujo, A. Lee, J. Zhang, L. Lee, S. H. Cheong, J. E. Duan, 2024 Transcriptomic difference of in vitro produced male and female early embryos in bovine. To be presented as a poster presentation at the 51st annual meeting of the International Embryo Technology Society, Fort Worth, TX</li><br /> <li>L. Murphy, A.K. Goldkamp, M.J.A. Lopes, N.F.F. Bonmann, M.C. Lucy, D.E. Hagen, J.G.N. Moraes. Alterations in miRNA and tRNA-derived fragment expression during bovine conceptus elongation. 2025 Annual Meeting of the International Embryo Technology Society (IETS), January 18-22, 2025, Fort Worth, Texas, USA.</li><br /> <li>Sellmer Ramos, M. O. Caldeira, S. E. Poock, Moraes, Joao G.N., Lucy, Matthew C, Patterson, Amanda L. Characterization of the endometrium epithelium of postpartum dairy cows using a multinomial transcriptomic approach to investigate disease programming of fertility. 21st Annual Gilbert S. Greenwald Symposium on Reproductive and Developmental Sciences, November 7-8, 2024, Kansas City, Kansas.</li><br /> <li>Roa-Vidal, Patricia, Markway, Macy, Moraes, Joao G.N., Lucy, Matthew C, Patterson, Amanda L. Uncovering the spatial gene expression of the bovine uterus: a novel study model for adenomyosis. 21st Annual Gilbert S. Greenwald Symposium on Reproductive and Developmental Sciences, November 7-8, 2024, Kansas City, Kansas.</li><br /> <li>J. Wilkinson, M.J.A. Lopes, G.L. Murphy, R.C. Botigelli, D. Londo&radic;&plusmn;o-V&radic;&deg;squez, Z. Jiang, A.C. Denicol, D.E. Hagen, J.G.N. Moraes. Establishment of protocols and conditions for culturing bovine embryonic stem cells. 2024 Poster Presentation of the Summer Research Training Program (SRTP) of the College of Veterinary Medicine, Oklahoma State University, August 8-10, Stillwater, OK.</li><br /> <li>F.F. B&radic;&part;nmann, L.G.D. Mendon&radic;&szlig;a, I. Sellmer Ramos, R. Fritz, C. Gamarra, D. Duhatschek, R.S.S. de Oliveira, A.L.A. Scanavez, T.S. Belem, M.C. Lucy, J.G.N. Moraes. Behavioral and physiological differences of dry dairy cattle under heat stress based on median core body temperature. The 2024 ADSA Annual Meeting is scheduled for June 16-19, 2024, in West Palm Beach, Florida.</li><br /> <li>Van Rhijn S., Murphy G.L., Goldkamp A., Lopes M.J.A., Bonmann N., Hagen D., Moraes J.G.N. Transcriptional Regulation of Bovine Conceptus Elongation. 2024 OSU Undergraduate Research Symposium, April 16th, 2024, Oklahoma State University, USA.</li><br /> <li>Buckmaster, K. L., Melton, C., Lopes, M.J.A., Lamberti G.S.E., Briggs E. A., Lalman D.L., Moraes J.G.N. The Role of Interferon-Stimulated Genes in Early Recognition of Pregnancy in Cattle. Wentz and Purdie Research Scholars Symposium, April 25th, 2024, Oklahoma State University, USA.</li><br /> <li>Buckmaster, K. L., Melton, C., Lopes, M.J.A., Lamberti G.S.E., Briggs E. A., Lalman D.L., Moraes J.G.N. The Role of Interferon-Stimulated Genes in Early Recognition of Pregnancy in Cattle. 2024 Research Day at the Capitol, Oklahoma State University, March 18-19, 2024, Oklahoma City, Oklahoma, USA.</li><br /> <li>H. McDonald, C. P. Johnson, T. P. Vining, M. D. Major, G.S.E. Lamberti, M.I.A. Lopes, N.F.F. Bonmann, J.G.N. Moraes, and P. A. Beck. Effects of sire selection on puberty and reproduction of offspring selected as replacement heifers. 2024 American Society of Animal Sciences (ASAS) Southern Section Meeting, January 27-30, 2024, Louisville, Kenticky, USA.</li><br /> <li>Lamberti G.S., Goldkamp A.K., Lopes, M.J.A., Bonmann, N.F. F., Van Rhijn S., Hagen, D.E., Moraes, JGN. Unraveling the biology of conceptus elongation in cattle. 2024 American Society of Animal Sciences (ASAS) Southern Section Meeting, January 27-30, 2024, Louisville, Kentucky, USA.</li><br /> <li>Culler H, Zarzaca I, Mueller V, Elgarresta C, Hersom MJ, Checura CM. Effect of photobiomodulation in blastocysts post-vitrification. Reproduction, Fertility and Development 2025: 37 (1) 41. https://doi.org/10.1071/RDv37n1Ab41</li><br /> <li>Elgarresta C, Mueller V, Zarzaca I, Culler H, Duckett SK, Klotz JL, Hersom MJ, Checura CM. Vasoconstriction induced by ergot alkaloids affects oocyte developmental capacity in pregnant sheep. Reproduction, Fertility and Development 2025: 37 (1) 195. https://doi.org/10.1071/RDv37n1Ab195</li><br /> <li>Elgarresta C, Kordus R, Erck S, Fusco A, Strawbridge EG, Watson R, Checura CM. The Effect of Different Spermatozoa Preparation Techniques on DNA Fragmentation and the Presence of Reactive Oxygen Species. Clemson University 8th Annual Summer CI + UR Showcase, Clemson, SC. 2024: 24.</li><br /> <li>Klotz JL, Duckett SK, Checura CM. Serotonin stimulates relaxation in ovine saphenous vein precontracted with ergovaline. American Society of Animal Sciences Annual Meeting, Calgary, AB, Canada: 2024: 226. https://doi.org/10.1093/jas/skae234.324</li><br /> <li>Elgarresta C, Mueller V, Zarzaca I, Culler H, Hersom M, Checura CM. The Central Role of Mitochondria in In Vitro Fertilization. Clemson University 19th Annual Focus on Creative Inquiry Forum, Clemson, SC. 2024: 190.</li><br /> <li>Culler H, Richey KD, Checura CM. Photobiomodulation during oocyte maturation enhances blastocyst rates but not blastocyst adenosine triphosphate content. Reproduction, Fertility and Development 2024: 36 (2) 259. https://doi.org/10.1071/RDv36n2Ab208</li><br /> <li>Thomason L, Wilhelm C, Mandel A, Kirkman K, Richey KD, Culler H, Hersom M, Checura CM. Photobiomodulation of bovine zygotes at 21 hours postfertilization. Reproduction, Fertility and Development, 2024: 36 (2), 197. https://doi.org/10.1071/RDv36n2Ab91</li><br /> <li>Luense, LJ Plant and Animal Genomics, San Diego, CA, January 12^th, 2024, Investigating the Effects of an Abnormal Sperm Histone Epigenome on Preimplantation Embryo Development</li><br /> <li>R. Dodd, L.J. Luense. Investigating the presence of acetylated histone post-translational modifications in bovine preimplantation embryos. Society for the Study of Reproduction 2024.</li><br /> <li>J. Luense, G. Donahue, M. Mendoza, N. Leu, B. Garcia, M.S. Bartolomei, S.L. Berger, Abnormal sperm histone signature leads to altered preimplantation embryogenesis. Gordon Research Conference &ndash; Mammalian Reproduction, Barcelona, Spain, July 2024</li><br /> <li>R. Dodd, L.J. Luense. Investigating the presence of acetylated histone post-translational modifications in bovine preimplantation embryos. Texas A&amp;M University Interdisciplinary Faculty of Reproductive Biology Retreat, Nov 2024</li><br /> <li>Adams, L., Liu, Y., Patrick, T., Grow, E., Ruggeri, E., Durrant, B. Polejaeva, I.A.* (2024). Quantification of mitochondrial DNA copy number in interspecies somatic cell nuclear transfer embryos. (2nd ed., vol. 35, pp. 136-136). Reproduction, Fertility and Development. 36:158-159.</li><br /> <li>Bennett, K., Beal, D., Liu, Y., Patrick, T., Bunderson, I., Moawad, A., Polejaeva, I.A.* (2024). Effects of oxygen level on bovine oocyte maturation and embryo development. Reproduction, Fertility and Development. 36:257.</li><br /> <li>Blocher, R., Liu, Y., Polejaeva, I.A.* (2024). Nuclear assessment of bovine oocytes matured in cytokine-supplemented medium using microtubule and DNA staining. Reproduction, Fertility and Development. 36:262.</li><br /> <li>Bunderson, I., Liu, Y., Polejaeva, I. A.* (2024). Effects of serum-free maturation medium and resveratrol supplementation on ovine oocyte maturation and quality. Reproduction, Fertility and Development. 36:263.</li><br /> </ol><br /> <p><strong>Miscellaneous publications (semi-technical/lay publications)</strong></p><br /> <ol><br /> <li>Looney, Charles R. (2024).&nbsp; Culling beef cows for profit.&nbsp; Arkansas Cattle Business. November 2024.</li><br /> <li>Looney, Charles R., Schowe, Clayton. (2024).&nbsp; Using Beef Genetics for Dairy Calf Production Increases Sustainability.&nbsp; Dairy E-News</li><br /> <li>Jiang Z. METTL7A improves embryo competence. 2024 Sep 24. U.S provisional patent application 63/698,174.</li><br /> <li>Jiang Z, Ming H. Methods and compositions for generating bovine extraembryonic endoderm cells and blastoids. 2024 Dec 16. US provisional patent application 63/734,491.</li><br /> <li>US Patent Number. 12,067,712B2 M.B. Wheeler et al. Date of Patent Issued :Aug. 20, 2024. Title: Phase Imaging with Computational Specificity&rdquo; Filed: February 18, 2020. Application: Assessment for viability of sperm and embryos via artificial intelligence neural networks.</li><br /> </ol><br /> <p>&nbsp;</p><br /> <p>&nbsp;</p>

Impact Statements

1. IMPACTS Objective 1 1. Results using the 14-day (8&6) protocol with insemination of cows at 66 and 72h post-CIDR removal with conventional or sexed female semen for the same bull has provided similar not-significantly different pregnancy rates. In addition, these projects have shown that delaying breeding is not needed when utilizing sexed-sorted semen. 2. Optical imaging offers a potential non-invasive and accurate method to determine viability of bovine embryos. Methods developed with bovine embryos can easily apply to the human IVF clinics. 3. Indistinguishable bESC lines can be readily derived from eBL and BL, widening the pool of embryos available for bESC establishment. 4. To investigate the impact of CRISPR-cas9-mediated knockout of Keap1 gene in modulating NRF2 activity, three guide RNAs targeting the exon 2 of the Keap1 gene have been tested for their efficiency in inducing targeted knockout of the gene in zygotes electroporated with RNP at 10 h post IVF. 5. Investigated the impact of granulosa cell derived stress associated extracellular vesicles supplementation during oocyte maturation under thermal stress condition. In that experiment we found that supplementation of EVs derived from thermal stress granulosa cells can induce tolerance to oocytes subjected to heat stress by reducing the accumulation of ROS, improving mitochondrial activity, reducing the expression of stress associated genes thereby reducing the severity of stress in oocytes and developing blastocysts. 6. Investigated the cellular level ovarian response to environmental seasonal changes in winter and summer season. 7. Used a bovine oviductal organoid 3D culture model to recapitulate the oviductal cellular and extracellular response to thermal stress. 8. Research on extracellular vesicles-mediated molecular signaling in ovarian follicle and oocytes in will facilitate the development of diagnostic markers associated with maternal physiology and embryo developmental competence and future potential therapeutic application of EVs in assisted reproductive technologies. 9. The use of bovine oviduct organoid technology will allow the generation of physiologically relevant EVs as molecular cargo in embryo oviduct communication for future application in ART to enhance in vitro embryo production yield and quality. 10. Results of the study on pathogens in bovine IVF systems were used in an assessment on safety of in vitro produced embryos that is currently being conducted by the Health and Safety Advisory Committee (HASAC) of the IETS. Further discussion of the data and next steps will be addressed at the IETS meeting in January 2025. The goal is to provide evidence-based import/export standards that are accessible to the global livestock industry. 11. In human IVF, pre-implantation genetic testing based on aneuploidy has not been found to increase pregnancy or live birth rates. Our data on ploidy correction by early embryos should shed lights on the long-held observation that the early embryos can correct its ploidy and return to the default of diploid. 12. Single-cell transcriptomic analysis of bovine blastocysts produced in vivo and in vitro in conventional and reduced nutrient conditions reveals the effect of culture environments on embryo developmental potential. 13. We have identified a novel mitochondria stress eliminating mechanism regulated by METTL7A that occurs during the acquisition of oxidative stress in embryo in vitro culture. This discovery lays the groundwork for the development of METTL7A as a promising therapeutic target for IVF embryo competence. 14. Our work is the first characterization of the mRNA translational spatiotemporal dynamics that accompanies mammalian pre-implantation development, and demonstration the necessity of a 2-cell-specific translated eukaryotic initiation factor, Eifad3, and its variants in pre-implantation development, demonstrating the significant value of this unprecedented resource for prioritizing genes for functional characterization. Thus, our work has filled a significant knowledge gap in the study of translational regulation and provided an extensive resource that can be mined for expanded insights into mammalian germ cell and preimplantation development. 15. Our work provides foundational information to discover essential biological pathways underpinning bovine pre- and peri-implantation development and the molecular causes of the early pregnancy failure during this critical period. 16. Our analysis reveals that both BAP and TSC models have features of peri-implantation TB s, while maintaining minor transcriptomic differences, and thus serve as valuable tools for studying implantation in lieu of human embryos. 17. Optimization of high resolution respirometry assays to assess mitochondrial bioenergetics of bull sperm have application for inclusion in semen quality assays that may strengthen the ability to predict fertility prior to insemination. These advances in complement with statistical models may help to reduce monetary losses associated with failed conception for the cattle industry. 18. Determination of postejaculatory sperm susceptibility to epigenetic modifications will likely create a paradigm shift in understanding how the uterine environment may alter sperm function and subsequent embryo development. 19. The effects of THC on postejaculatory sperm shed new light to negative impacts of marijuana use on sperm function. These studies likely provide the most comprehensive assessment to date while serving as a platform for understanding impacts on fertility and embryo development. 20. Optimization of conditions for stallion sperm that are conducive to equine IVF will contribute to protocols that can be improved for generating equine IVF embryos. These findings have the potential to positively influence commercial applications for equine IVF by improving efficiency of methods while contributing toward fundamental knowledge of stallion sperm physiology. 21. Expanding the shelf-life of cooled stallion sperm for artificial insemination may influence industry standards to improve transport devices that maintain internal temperatures for longer durations. Such applications would greatly relieve logistical constraints of artificial insemination with stallion sperm and positively influence industry application by reducing shipment costs, the frequency of stallion collections, and reducing intensive management of both the stallion and mare. 22. Complementation approaches to generate identical bovine embryos from a single 4-cell embryo may impact industry methods that circumvent the need for low-efficiency cloning strategies while minimizing technical expertise required for current practices. 23. First characterization of peroxisome-proliferator activated gamma expression in bovine embryos suggests a role in early embryonic development that may be leveraged to improve in vitro culture conditions and optimize embryo quality. 24. Showed that heat-stress (HS) can compromise the recruitment of ovarian follicles early in a new follicular wave in Holstein dairy cattle. 25. We provided the first report embryonic gene expression changes due to fescue endophyte (E+) exposure before fertilization in beef cattle. It may provide novel insights to mitigate the effects of fescue toxicosis on beef reproduction efficiency. 26. Feeding rumen-protected methionine during the transition period and early lactation altered specific lipid classes and lipid unsaturation level of preimplantation embryos and endometrial tissue. 27. Time-lapse monitoring and artificial-intelligence-based automated image analysis have the potential for accurate embryo evaluation; however, further research is warranted to innovate economically feasible options for in-field applications. 28. Characterization of the transcriptome, molecular fingerprint and mitochondrial function between conventional in vitro maturation and extended maturation will provide clues for the improvement of this key assisted reproductive technology. 29. Manipulating cathepsin V level in early embryos is a promising approach to improve the efficiency of IVP technology in cattle. 30. Identification and flux quantification of the novel sulfur amino acid-based redox homeostasis mechanisms revealed previously unknown mechanisms cells can use to support redox homeostasis. These are likely to be important in embryonic development, which we plan to investigate in future studies. 31. Establish protocols to profile the epigenetic regulator MOF and its associated marker H4K16ac during bovine embryonic development. This fills a fundamental knowledge gap by establishing protocols to profile an epigenetic regulator MOF and H4K16ac during bovine embryonic development. 32. Investigate the mechanisms driving developmental rate differences between male and female embryos. This will identify the underlying mechanisms behind the developmental rate difference between male vs. female embryos. 33. Characterize the functional role of transposable elements in bovine early development. This will understand the functional role of transposon elements during bovine early development. 34. Projects will provide valuable insights into embryonic epigenetics and offer a unique opportunity to develop a targeted biomarker assessment system. This system has the potential to enhance fertility and reproductive efficiency, addressing a critical challenge for the cattle industry. 35. The novel microRNA and tRNA-derived fragment (tRF) data generated during bovine preimplantation development provide valuable insights into the developmental biology processes of early embryogenesis in cattle. The long-term goal of this research is to improve cattle production systems and alleviate the impact of reproductive challenges in cattle operations. 36. By using photobiomodulation, the role of mitochondria at the different developmental stages is better understood. 37. Determining that H3K27ac is located at transcription factor binding sites that are required for zygotic gene activation, and cell differentiation provide key insight into how paternal histones regulate early embryogenesis. 38. Improvement in proper meiotic spindle configuration at MII stage in bovine oocytes, and accelerated the SCNT embryo development derived from FLI-supplemented IVM could have a significant impact on improving development and quality of embryo derived from in vitro production, and animal production. 39. The identified parameters for SCNT embryo production would enhance SCNT animal production for both agricultural and medical applications. 40. Identifying the expression pattern of CPEB1 during early embryo development would be beneficial for understanding its function at each stage in the embryos and could shed the light on potential use of CPEB1 to improve in vitro embryo production. 41. Although CoQ10 does not affect oocyte quality and blastocyst formation, increased mitochondrial mass in oocytes may potentially improve in vivo embryo development. Objective 2 1. Although preantral follicles can initiate growth without the hormone, FSH signaling plays a crucial role in their optimal development from an early stage. 2. Therefore, production of TTR by the placenta may be necessary for the delivery of TH to the fetus however, this theory has not been tested in vivo. 3. While numerous methods have been tested to separate bovine X- and Y-bearing sperm, most of them are either unpractical or have failed in repeatability. We tried to confirm a reported new method and started the question on the value of this sperm separation technology. 4. In the bovine generating genetically modified animals have not been achieved with the use of tetraploid complementation. Our study on triploid complementation may be a good alternative approached to generate transgenic animals in one generation. 5. The development of an alternative method for delivering DNA constructs to bovine embryos is a promising approach as a more efficient technology for intentional genetic alterations of livestock. We anticipate that further optimization will result in translational applications for producing animals of high genetic merit and as a tool to reduce the frequency of deleterious alleles. 6. We report the derivation and long-term culture of bovine extraembryonic endoderm cells (XENs) from pre-implantation embryos. Importantly, this study not only demonstrates the utility of bovine XEN models in elucidating the mechanistic features of early bovine embryogenesis, but also develops an improved bovine blastocyst-like structures (blastoids) technology for the creation of novel assisted reproductive technologies. 7. For the first time, it was demonstrated that mature bioactive human insulin can be produced in cows milk, which could potentially revolutionize how insulin is administered to diabetic patients. 8. Improved outcomes from bovine intra cytoplasmic sperm injection will provide an additional method for incorporation of gene editing techniques and fertilization with spermatozoa for which there are limited numbers such as with sex sorted sperm. 9. Rapid detection of off-targeting events can improve safety of gene editing technology. 10. Use of Cas9 variants can support precise gene editing. 11. The AAV8-CRISPR-Cas9 liver-targeting genome editing systems developed here provide novel tools, assays, and approaches that not only validate and advance the redox systems at hand now, but could also help improve these types of tools for use in livestock. 12. Embryonic stem cells (ESC) are pluripotent cells derived from the inner cell mass (ICM) of blastocysts, with the potential to differentiate into most cell types, including gametes, given appropriate conditions. The development of stable conditions for culturing and differentiation of bovine and bison ESC holds significant potential in veterinary medicine (e.g., regenerative medicine) and for accelerating genetic improvements, particularly when combined with assisted reproductive technologies (ART). 13. The established cell lines can be utilized in future somatic cell nuclear transfer experiments for generating genetically engineered pig models intended for xenotransplantation research. 14. Improving mitochondria efficiency enhances the in vitro embryo production system and associated methodologies. 15. Parameters were identified to improve SCNT outcomes for the production of genetically engineered livestock.

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