Apoptotic and proliferative changes during induced atresia of pre-ovulatory follicles in the rat
Atresia, a degenerative process through which many follicles are removed from the growing pool, involves apoptotic changes in the follicular granulosa cells. To identify histochemical markers of early stages of atresia, an in-vivo rat model was used which allowed the study of atresia of pre-ovulatory follicles in a synchronized and chronological order. By blocking the pre-ovulatory luteinizing hormone surge with a gonadotrophin-releasing hormone (GnRH) antagonist, ovulation of the pre-ovulatory follicles is prevented, after which these follicles became atretic. The first morphological sign of atresia (pyknotic granulosa cell nuclei) was found 27 h after injection of GnRH antagonist. Since the pre-ovulatory follicles gradually become atretic in a synchronous fashion, this model provided an opportunity to study and define markers of future atresia in pre-ovulatory follicles. Atresia involves apoptosis of granulosa cells, and therefore internucleosomal DNA fragmentation was examined. Using the terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labelling (TUNEL) assay it was found that the first sign of internucleosomal DNA fragmentation in granulosa cells of pre-ovulatory follicles was detectable 24 h after GnRH antagonist treatment. In order to find an upstream marker of atresia, the 5-bromo-deoxyuridine (BrdU) labelling index was used as a measure of proliferation. Already at 14 h after GnRH antagonist treatment, when morphological signs of atresia were not yet present, a clear decrease in BrdU labelling index was found in the granulosa cells.
|Keywords||*Apoptosis, *Cell Division, *Follicular Atresia, *Ovulation/drug effects, Animals, Bromodeoxyuridine/metabolism, Cell Nucleus/ultrastructure, DNA Fragmentation, Female, Gonadorelin/antagonists & inhibitors, Granulosa Cells/ultrastructure, Immunohistochemistry, In Situ Nick-End Labeling, Luteinizing Hormone/secretion, Meiosis, Oocytes/cytology, Proestrus, Rats, Rats, Wistar|
Durlinger, A.L.L., Kramer, P., Karels, B., Grootegoed, J.A., Uilenbroek, J.Th.J., & Themmen, A.P.N.. (2000). Apoptotic and proliferative changes during induced atresia of pre-ovulatory follicles in the rat. Human Reproduction. Retrieved from http://hdl.handle.net/1765/9546