Regulation of apoptosis in the atresia of dominant bovine follicles of the first follicular wave following ovulation

• Published in: Reproduction (Cambridge, England) Vol. 130; no. 1; pp. 71 - 81
• Main Authors: Valdez, Kelli E, Cuneo, S Peder, Turzillo, Adele M
• Format: Journal Article
• Language: English
• Published: Colchester Society for Reproduction and Fertility 01.07.2005; Portland
• Subjects: Animals; Annexin A5; apoptosis; Apoptosis - genetics; Aromatase - analysis; Biological and medical sciences; Blotting, Western - methods; Caspase 3; Caspase 8; Caspase 9; Caspases - analysis; Cattle; Coloring Agents; dairy cows; Estradiol - blood; Female; follicular atresia; Follicular Atresia - physiology; follicular development; Fundamental and applied biological sciences. Psychology; Genes, bcl-2; granulosa cells; Granulosa Cells - chemistry; heifers; Hormone metabolism and regulation; Mammalian female genital system; Ovarian Follicle - metabolism; ovarian follicles; ovulation; Progesterone - blood; Propidium; Reactive Oxygen Species - analysis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - analysis; Superoxide Dismutase - genetics; Theca Cells - metabolism; Vertebrates: reproduction; Ox; Ovulation; Ovary; Vertebrata; Reproduction; Mammalia; Cell death; Animal; Female genital system; Atresia; Artiodactyla; Ovarian follicle; Ungulata; Apoptosis
• ISSN: 1470-1626; 1741-7899
• DOI: 10.1530/rep.1.00430

During atresia of bovine follicles, granulosa cells are lost through the controlled form of cell death, apoptosis. The purpose of this study was to characterize the regulation of apoptotic death of granulosa cells in dominant bovine follicles during the first wave of follicular development. Dominant follicles were collected from Holstein heifers on days 4, 6 or 8 of the first follicular wave (n = 5/day). Regulation of apoptosis in granulosa cells was examined by annexin V and propidium iodide staining; measurement of relative levels of mRNA encoding Bcl-2, Bcl-xL and Bax; and activity of caspase-3, -8 and -9. Steady-state levels of mRNA encoding four oxidative stress-response proteins were determined. Compared with day 4, the incidence of apoptotic and nonviable granulosa cells tended to increase on day 6, and numbers of nonviable cells were higher on day 8. The ratios of relative levels of mRNA encoding Bcl-2 to Bax and Bcl-xL to Bax were higher on day 6 than days 4 and 8. Activity of caspases-3 and -9 in granulosa cells did not change among the 3 days, while caspase-8 activity decreased on day 8 compared with days 4 and 6. Amounts of GSHPx, MnSOD and Cu/ZnSOD mRNA in granulosa cells were higher on day 8 than day 6. In theca interna, amounts of Cu/ZnSOD mRNA decreased between days 4 and 6. From the decreased production of estradiol and increased numbers of apoptotic and nonviable granulosa cells, we conclude that atresia of the dominant follicle is initiated between days 4 and 6 of the first follicular wave. However, apoptosis of granulosa cells does not appear to be initiated by changes in expression of oxidative stress-response proteins.