Follicular expression of a human β-cell leukaemia/lymphoma-2 (Bcl-2) transgene does not decrease atresia or increase ovulation rate in swine*
H. D. Guthrie A C , R. J. Wall A , V. G. Pursel A , J. A. Foster-Frey A , D. M. Donovan A , H. D. Dawson B , G. R. Welch A and W. G. Garrett AA Biotechnology and Germplasm Laboratory, Animal and Natural Resources Institute, Agricultural Research Service, USDA, Beltsville, MD 20705, USA.
B Nutrient Requirement and Functions Laboratory, Beltsville Human Nutrition Research Center, Agricultural Research Service, USDA, Beltsville, MD 20705, USA.
C Corresponding author. Email: dave@anri.barc.usda.gov
Reproduction, Fertility and Development 17(4) 457-466 https://doi.org/10.1071/RD04136
Submitted: 16 November 2004 Accepted: 27 February 2005 Published: 12 April 2005
Abstract
Transgenic (TG) gilts carrying a human Bcl-2 cDNA transgene driven by mouse inhibin-α subunit promoter were produced and evaluated to determine if ectopic expression of Bcl-2 in the ovaries would decrease the frequency of atresia in antral follicles and increase ovulation rate. Immunohistochemical analysis showed that the Bcl-2 transgene protein was expressed in granulosa and theca cells, in 86% of healthy and 54% of atretic follicles analysed in TG prepubertal and Day 50 pregnant gilts combined (n = 24). In contrast, Bcl-2 transgene protein was expressed in only 1.4% of healthy and 0% of atretic follicles in non-TG littermates (n = 13). Real-time reverse transcription–polymerase chain reaction analysis confirmed that human Bcl-2 was expressed in follicles of TG gilts. The atresia rate for the TG and non-TG groups did not differ (P > 0.05) for prepubertal (45 v. 59%) and Day 50 pregnant gilts (53 v. 52%) respectively. The mean ± s.e.m. ovulation rate did not differ (P > 0.5) between TG (15.9 ± 0.8, n = 12) and non-TG (16.4 ± 0.6, n = 7) Day 50 pregnant gilts. The molecular basis of the failure of ectopic Bcl-2 expression to increase the ratio of healthy to atretic follicles is unknown, but it is possible that the activity of the mitochondrial-dependent cell death pathway was not neutralized by ectopic expression of human Bcl-2 or that other cell death pathways compensated for the decreased mitochondrial-dependent cell death.
Acknowledgments
We thank Leah L. Schulman, Delbert Parsons and James Piatt for their technical assistance.
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