Evidence for ovarian granulosa stem cells: telomerase activity and localization of the telomerase ribonucleic acid component in bovine ovarian follicles

• Published in: Biology of reproduction Vol. 61; no. 2; pp. 358 - 366
• Main Authors: Lavranos, T.C, Mathis, J.M, Latham, S.E, Kalionis, B, Shay, J.W, Rodgers, R.J
• Format: Journal Article
• Language: English
• Published: Madison, WI Society for the Study of Reproduction 01.08.1999
• Subjects: Animals; Biological and medical sciences; Cattle; Cell Cycle; Cell Line; Digoxigenin - metabolism; embryonic stem cells; enzyme activity; Female; Flow Cytometry; follicle size; Fundamental and applied biological sciences. Psychology; Graafian follicles; granulosa cells; Granulosa Cells - chemistry; Granulosa Cells - enzymology; Humans; In Situ Hybridization; Ki-67 Antigen - biosynthesis; localization; Mammalian female genital system; Morphology. Physiology; Ovarian Follicle - cytology; Ovarian Follicle - enzymology; RNA; RNA - metabolism; RNA, Long Noncoding; RNA, Untranslated; RNA-directed DNA polymerase; Telomerase - metabolism; Vertebrates: reproduction; Cell culture; Enzyme; Transferases; Stem cell; Ox; Gene expression; Ovary; Nucleotidyltransferases; Vertebrata; Mammalia; Enzymatic activity; Female genital system; Artiodactyla; Granulosa; Ovarian follicle; Ungulata; DNA-directed DNA polymerase
• ISSN: 0006-3363; 1529-7268
• DOI: 10.1095/biolreprod61.2.358

We have previously postulated that granulosa cells of developing follicles arise from a population of stem cells. Stem cells and cancer cells can divide indefinitely partly because they express telomerase. Telomerase is a ribonucleoprotein enzyme that repairs the ends of telomeres that otherwise shorten progressively upon each successive cell division. In this study we carried out cell cycle analyses and examined telomerase expression to examine our hypothesis. Preantral (60–100 μm) and small (1 mm) follicles, as well as granulosa cells from medium-sized (3 mm) and large (6–8 mm) follicles, were isolated. Cell cycle analyses and expression of Ki-67, a cell cycle-related protein, were undertaken on follicles of each size (n = 3) by flow cytometry; 12% to 16% of granulosa cells in all follicles were in the S phase, and less than 2% were in the G 2 /M phase. Telomerase activity (n = 3) was highest in the small preantral follicles, declining at the 1-mm stage and even further at the 3-mm stage. In situ hybridization histochemistry was carried out on bovine ovaries, and telomerase RNA was detected in the granulosa cells of growing follicles but not primordial follicles. Two major patterns of staining were observed in the membrana granulosa of antral follicles: staining in the middle and antral layers, and staining in the middle and basal layers. No staining was detected in oocytes. Our results strongly support our hypothesis that granulosa cells arise from a population of stem cells.