CRIS NUMBER: 0197687
SUBFILE: CRIS
PROJECT NUMBER: VA-135707
SPONSOR AGENCY: CSREES
PROJECT TYPE: HATCH
PROJECT STATUS: TERMINATED
MULTI-STATE PROJECT NUMBER: (N/A)
START DATE: Oct 1, 2003
TERMINATION DATE: Sep 30, 2008
GRANT PROGRAM: (N/A)
GRANT PROGRAM AREA: (N/A)
CLASSIFICATION
301 | 3310 | 1020 | 2.2 | 10% |
301 | 3310 | 1030 | 2.2 | 10% |
301 | 3310 | 1040 | 2.2 | 10% |
301 | 3310 | 1050 | 2.2 | 10% |
301 | 3410 | 1020 | 2.2 | 10% |
301 | 3410 | 1030 | 2.2 | 10% |
301 | 3410 | 1040 | 2.2 | 10% |
301 | 3410 | 1050 | 2.2 | 10% |
301 | 3510 | 1030 | 2.2 | 10% |
301 | 3510 | 1050 | 2.2 | 10% |
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CLASSIFICATION HEADINGS
KA301 - Reproductive Performance of Animals S3310 - Beef cattle, live animal S3510 - Swine, live animal S3410 - Dairy cattle, live animal F1020 - Physiology F1030 - Cellular biology F1050 - Developmental biology F1040 - Molecular biology G2.2 - Increase Efficiency of Production and Marketing Systems
RESEARCH EFFORT CATEGORIES
BASIC |
50% |
APPLIED |
25% |
DEVELOPMENTAL |
25% |
KEYWORDS: in vitro; abnormalities; sperm; embryos; embryo development; fertilization; beef cattle; dairy cattle; reproductive performance; swine; animal physiology; cell biology; molecular biology; embryo survival; biotechnology; new technology; spermatozoa; zygotes; cleavage; apoptosis; cell morphology; semen; reproductive efficiency
PROGRESS: Oct 1, 2006 TO Sep 30, 2007
OUTPUTS: Normal embryonic development depends on the maintenance of a population of normal healthy cells within each embryo. Thus, the disposal of morphologically abnormal cells and cells in excess is crucial during early development. Studies were conducted to evaluate the effect of vitrification of oocytes and early embryos on embryonic development after IVF and to use a combination of apoptotic (programmed cell death) measures to assess differences in embryo quality. The concept of ultra-rapid vitrification has emerged in recent years; the accelerated cooling rate reduced injury attributed to cryopreservation and improved post-freezing developmental competence of vitrified oocytes and embryos. The objectives of the present study were to develop a simple and effective ultra-rapid vitrification method (droplet vitrification) and evaluate its effects on post-thaw development and apoptosis-related gene expression in mouse zygotes. Presumptive zygotes were equilibrated for 3 min in
equilibration medium and washed 3 times in vitrification solution. A drop (5 mL) of vitrification solution containing 10-12 embryos was placed directly onto surface of liquid nitrogen, with additional liquid nitrogen poured over the drop. For thawing and cryoprotectant removal, vitrified drops were put into dilution medium for 3 min, followed by M2 medium for 5 min. Although cleavage rate did not differ significantly among the control (90.8%), toxicity control (83.5%), and vitrified (86.2%) zygotes, rates of blastocyst and hatched blastocyst formation were lower in vitrified zygotes (49.7% and 36.0%) and toxicity controls (47.3% and 40.3%) compared with controls (65.5% and 54.2%). Exposure of zygotes to vitrification solution, as well as the vitrification process, down-regulated the expression of Bax, Bcl2, and p53 genes in blastocysts. Although droplet vitrification was efficient and easy, it altered the transcriptional activities of Bax, Bcl2, and p53 genes in vitrified embryos,
indicating a strong relationship between reduced developmental competence and the altered transcriptional activities of these genes. Holstein cows were administered zona pellucida (ZP) DNA vaccine and used to determine the potential of recombinant rabbit ZP glycoproteins (rZP) as immunocontraceptive antigens. Cows were assigned to 1 of 4 treatment groups in which plasmids encoding rabbit ZP proteins were administered, i.d., using a gene gun (ZP55, n = 2; ZP75, n = 2; Hep55, n = 2; and Control, n = 3). Blood samples were taken before initial vaccination, once weekly for 5 wk and at 148 wk post-immunization. An ELISA was developed to assess anti-ZP titer levels in cow serum and ovarian function in cows was monitored using trans-rectal ultrasonography. Four of the six cows in ZP treatment groups developed antibody titer levels with similar linear responses over time. These cows also experienced reduced ovarian function as indicated by decreases in follicular and luteal activity. Estrous
activity was observed in all cows and decreased in ZP treatment cows in comparison to Controls. Further research is needed to determine the relationship between ZP immunocontraception and ovarian function.
PARTICIPANTS: Dhali, A., Visiting Professor; Anchamparuthy, V.M., Graduate Student; Butler, S.P., Collaborator; Pearson, R.E., Statistical consultant; Mullarky, I.K., Collaborator; Gwazdauskas, F.C., PI; Foley, C. A., Graduate Student; Boyle, S.M., Collaborator; Wilkes, C. O., Graduate Student; Pence, K. J., Graduate Student; Hurt, A. M., Graduate Student; Becvar, O., Collaborator; Knowlton, K. F., Collaborator; Mcgilliard, M. L., Statistical Consultant; Van Cott, K., Collaborator; Gil, G., Collaborator; Pipe, S. W., Collaborator; Miao, H. Z., Collaborator; Kaufman, R. J., Collaborator; Velander, W. H. Collaborator; Jones, E. T., Graduate Student; Guill, S. G., Graduate Student;Hargens, T.A., Graduate Student; Aron, A., Graduate Student; Butner, K. L., Graduate Student; Mabry, J. E., Graduate Student; Herbert, W. G., Collaborator;
TARGET AUDIENCES: Target audiences include individuals, groups, market segments in academia and the dairy and livestock industry. These groups include populations such as racial and ethnic minorities and those who are socially, economically, or educationally disadvantaged. Our data have been delivered as science-based knowledge to people through formal or informal educational programs such as: formal classroom instruction, laboratory instruction, or practicum experiences; workshops; and national meetings.
IMPACT: 2006-10-01 TO 2007-09-30
Rapid freezing altered the transcriptional activities of Bax, Bcl2, and p53 genes in vitrified embryos, indicating a strong relationship between reduced developmental competence and the altered transcriptional activities of these genes. Further research is needed to determine the relationship between ZP immunocontraception and ovarian function.
PUBLICATION INFORMATION: 2006-10-01 TO 2007-09-30
Dhali, A., Anchamparuthy, V.M., Butler, S.P., Pearson, R.E., Mullarky, I.K., Gwazdauskas, F.C. (2007) Gene expression and development of mouse zygotes following droplet vitrification. Theriogenology 68: 1292-1298.
PROJECT CONTACT INFORMATION
NAME: |
Gwazdauskas, F. C. |
PHONE: |
540-231-4756 |
FAX: |
540-231-5014 |
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