GERM CELL AND EMBRYO DEVELOPMENT AND MANIPULATION FOR THE IMPROVEMENT OF LIVESTOCK

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 1021181
• Multistate No.: W-4171
• Project Start Date: Jan 21, 2020
• Project End Date: Sep 30, 2024
• Program Code: [(N/A)]- (N/A)

Recipient Organization
LOUISIANA STATE UNIVERSITY
202 HIMES HALL
BATON ROUGE,LA 70803-0100

Performing Department
School of Animal Science

Non Technical Summary
Significant research effort concerning improved reproduction of animals is occurring at public institutions and in private companies and it is imperative that research of this type be publicly funded to ensure availability of results to animal agriculture and to private industry. This is particularly important for the Louisiana animal agriculture industry since it is centered around the breeding and marketing of genetically elite animals. Combining technologies such as in vitro fertilization, nuclear transfer, and cryopreservation provides phenomenal opportunities for increased efficiency of livestock production. In vitro embryo production could be timed to be used to increase flexibility in animal production. Oocytes from genetically elite females could be betterutilized, leading to a faster rate of genetic improvement due to increased selection intensity.

Animal Health Component

(N/A)

Research Effort Categories

Basic

70%

Applied

(N/A)

Developmental

30%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
301	3310	1020	20%
303	3310	1040	40%
305	3310	1030	40%

Knowledge Area
303 - Genetic Improvement of Animals; 305 - Animal Physiological Processes; 301 - Reproductive Performance of Animals;

Subject Of Investigation
3310 - Beef cattle, live animal;

Field Of Science
1040 - Molecular biology; 1030 - Cellular biology; 1020 - Physiology;

Keywords

bovine

oocyte maturation

cryopreservation

epigenetic marks

gene expression

Goals / Objectives
Understand the biology of gamete development, fertilization and embryogenesis including the underlying cellular and molecular mechanisms. Refine methods to produce animals by genetic engineering or genome editing for the improvement of livestock production efficiency and development of human biomedical models.

Project Methods
Investigation of factors associated with development of the oocyte to increase efficiency of in vitro embryo production systems. Identification of markers for developmental competence of oocytes in livestock species.Evaluate the biology associated with meiotic and morphological changes that occur during cytoplasmic maturation of oocytes leading to developmental competence.Examine the effects of in vitro manipulation procedures, including cryopreservation, on epigenetic mechanisms and gene expression during development.Collaboration between member projects members by sharing of resources, protocols and technology.

Progress 01/21/20 to 09/30/20

Outputs
Target Audience:Farm animal producers and reproductive physiologists in industry and acedemia. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Results from these studies were included in one MS thesis completed in 2020. Results from these studies will also be included in a second MS thesis to be completed in early 2021. These projects provided opportunities for professional development and specific research training. How have the results been disseminated to communities of interest?Presentation at international scientific meetings and publication in journals. What do you plan to do during the next reporting period to accomplish the goals?For major goal 1: We plan to study the effect of cryopreservation of bovine oocytes and embryos on epigenetic mechanisms of gene expression control. We will also study the lipid profile of porcine and bovine oocytes and how these profiles change during oocyte maturation both in vitro and in vivo. For major goal 2: We will continue to study spindle repair following vitrification of bovine oocytes and the role of glutathione accumulation during maturation on this process. We will continue to study male pronuclei formation in bovine zygotes following fertilization by intracytoplasmic sperm injection. Specifically, we will study how different in vitro oocyte maturation conditions affect this process.

Impacts
What was accomplished under these goals? For major goal 1: It was determined that vitrification at the blastocyst stage resulted in altered gene expression in Day 14 elongated embryos after thawing and an additional 7-day culture in vitro. Pathway analyses suggest major effects on mitochondria function and energy production. For major goal 2: It was determined that a pretreatment of sperm with progesterone improved the rate of male pronuclei formation following fertilization of bovine oocytes by intracytoplasmic sperm injection. It was also shown that an extended post thaw incubation improved spindle reformation in vitrified bovine oocytes. Attempts to reduce spindle disruption in vitrified bovine oocytes with microtubule stabilizing agents resulted in the formation of micronuclei.

Publications

• Type: Journal Articles Status: Published Year Published: 2020 Citation: Ming H, Sun J, Pasquariello R, Gatenby L, Herrick J, Yuan Y, Pinto C, Bondioli KR, Krisher RL, Jiang Z. 2020. The landscape of accessible chromatin in bovine oocytes and early embryos. Epigenetics. 2020 Jul 14. PMID: 32663104. DOI: 10.1080/15592294.2020.1795602
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Li Y, Sun J, Ling Y, Hao M, Chen Z, Fang F, Liu Y, Cao H, Ding J, Cao Z, Zhang X, Bondioli K, Jiang Z, Zhang Y 2020. Transcriptional profiles of goat in vivo preimplantation embryo development. Reproduction, Fertility and Development. 2020 32:714-725.
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: L. Gatenby and K.R. Bondioli 2020. The effect of biological extenders on in vitro induction of the acrosome reaction in bovine spermatozoa. Reproduction Fertility and Development. 32:183.
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Gutierrez E., Jiang Z., Bondioli K. 2020. Extended culture after vitrification-warming helps in spindle recovery of bovine oocytes. Reproduction, Fertility and Development 32, 144.
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Jiang Z., Gutierrez E., Ming H., Foster B., Gatenby L., Mak C., Pinto C., Bondioli K. 2020. Effect of vitrification on global gene expression dynamics of bovine elongating embryos. Reproduction, Fertility and Development 32, 141.
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Ming H., Sun J., Pasquariello R., Herrick J. R., Yuan Y., Gutierrez E., Gatenby L., Bondioli K. R., Krisher R. L., Jiang Z. 2020. The landscape of accessible chromatin in bovine oocytes and early embryos. Reproduction, Fertility and Development 32, 125.
• Type: Theses/Dissertations Status: Published Year Published: 2020 Citation: Sperm Decondensation and Male Pronuclear Formation in Bovine Intracytoplasmic Sperm Injection. Louisiana State University Masters Thesis. Lauren Gatenby. 2020.