Cell proliferation and survival mechanisms underlying the abnormal persistence of follicular cysts in bovines with cystic ovarian disease induced by ACTH.

Cystic ovarian disease (COD) is an important cause of infertility that affects cattle. Alterations in the ovarian micro-environment of females with follicular cysts could alter the normal processes of proliferation and programmed cell death in ovarian cells. Thus, the objective in the present study was to evaluate apoptosis and proliferation in induced ovarian cystic follicles in cows to investigate the follicular persistence. Stage of estrous cycle was synchronized in 10 heifers and 5 were then subjected to the induction of COD by administration of ACTH. After the ovariectomy number of in situ apoptotic cells by TUNEL assay, active caspase-3, FAS/FASLG and members of the BCL2 family were compared by immunohistochemistry and multiplex PCR and cell proliferation by evaluation of Ki-67 protein and cyclin D1 and E mRNA. Significantly (p<0.05) lesser proliferative and apoptotic rates were found in cystic follicles from cows with COD compared with those with regular cycles. The relatively minimal proliferation found by immunohistochemistry with Ki-67 marker were confirmed by the gene expression of cyclin D1 and E. Lesser apoptotic rates were associated with decreased amounts of apoptotic-related proteins BAX, FASLG and caspase-3 as well as the in situ apoptosis detected by TUNEL assay, and increased amounts of the anti-apoptotic survival factor cellular BCL2 in the cystic follicles of the COD group. The BAX/BCL2 gene expression profile confirmed the immunohistochemical findings. Results from the present study indicate that cellular proliferation and apoptosis are altered in cystic follicles of cattle. The present study provides new insights into the molecular mechanisms underlying the aberrant persistence of follicular cysts and related diseases.

[1]  J. Tilly.  Apoptosis and ovarian function. , 1996, Reviews of reproduction.

[2]  E. Gimeno,et al.  Characterization of cytoskeletal proteins in follicular structures of cows with cystic ovarian disease. , 2007, Journal of comparative pathology.

[3]  J. E. Fortune,et al.  Ovarian follicular dynamics during the estrous cycle in heifers monitored by real-time ultrasonography. , 1988, Biology of reproduction.

[4]  D. Lubahn,et al.  Dominant bovine ovarian follicular cysts express increased levels of messenger RNAs for luteinizing hormone receptor and 3 beta-hydroxysteroid dehydrogenase delta(4),delta(5) isomerase compared to normal dominant follicles. , 2001, Biology of reproduction.

[5]  N. Isobe,et al.  Immunocytochemical study of cell proliferation in the cystic ovarian follicles in cows. , 2000, Theriogenology.

[6]  A. Johnson,et al.  Intracellular mechanisms regulating cell survival in ovarian follicles. , 2003, Animal reproduction science.

[7]  H. Ortega,et al.  The role of ACTH in the pathogenesis of polycystic ovarian syndrome in rats: hormonal profiles and ovarian morphology. , 2007, Physiological research.

[8]  M. Skinner,et al.  Developmental and hormonal regulation of keratinocyte growth factor expression and action in the ovarian follicle. , 1998, Endocrinology.

[9]  Marylène Lejeune,et al.  Quantification of diverse subcellular immunohistochemical markers with clinicobiological relevancies: validation of a new computer‐assisted image analysis procedure , 2008, Journal of anatomy.

[10]  G. Adams,et al.  Progress in understanding ovarian follicular dynamics in cattle. , 2008, Theriogenology.

[11]  S. Ben‐Sasson,et al.  Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation , 1992, The Journal of cell biology.

[12]  D. Porter,et al.  Expression and Activity of the Fas Antigen in Bovine Ovarian Follicle Cells1 , 2000, Biology of reproduction.

[13]  E. Gimeno,et al.  Estrogen receptors alpha and beta and progesterone receptors in normal bovine ovarian follicles and cystic ovarian disease. , 2007, Veterinary pathology.

[14]  E. Gimeno,et al.  Estrogen Receptors α and β and Progesterone Receptors in Normal Bovine Ovarian Follicles and Cystic Ovarian Disease , 2007 .

[15]  J. Kim,et al.  Biochemical and morphological identification of ceramide-induced cell cycle arrest and apoptosis in cultured granulosa cells. , 1999, Tissue & cell.

[16]  A. Hsueh,et al.  Ovarian follicle atresia: a hormonally controlled apoptotic process. , 1994, Endocrine reviews.

[17]  A. T. Peter.  An update on cystic ovarian degeneration in cattle. , 2004, Reproduction in domestic animals = Zuchthygiene.

[18]  A. Johnson,et al.  Involvement of apoptosis in ovarian follicular atresia and postovulatory regression. , 1991, Endocrinology.

[19]  E. Gimeno,et al.  Insulin-like growth factor I in sera, ovarian follicles and follicular fluid of cows with spontaneous or induced cystic ovarian disease. , 2008, Research in veterinary science.

[20]  J. Tilly,et al.  Gonadotropin suppression of apoptosis in cultured preovulatory follicles: mediatory role of endogenous insulin-like growth factor I. , 1994, Endocrinology.

[21]  R. Harman,et al.  Susceptibility of ovarian granulosa cells to apoptosis differs in cells isolated before or after the preovulatory LH surge , 2001, Molecular and Cellular Endocrinology.

[22]  H. Ortega,et al.  Microscopic characterization of follicular structures in letrozole-induced polycystic ovarian syndrome in the rat. , 2006, Archives of medical research.

[23]  D. Lubahn,et al.  Dominant Bovine Ovarian Follicular Cysts Express Increased Levels of Messenger RNAs for Luteinizing Hormone Receptor and 3β-Hydroxysteroid Dehydrogenase Δ4,Δ5 Isomerase Compared to Normal Dominant Follicles , 2001 .

[24]  H. Ortega,et al.  Insulin-like growth factor-II and insulin-like growth factor-binding proteins in bovine cystic ovarian disease. , 2010, Journal of comparative pathology.

[25]  D. Modi,et al.  Exposure of adult rats to estradiol valerate induces ovarian cyst with early senescence of follicles , 2007, Molecular and Cellular Endocrinology.

[26]  Hormone-induced proliferation and differentiation of granulosa cells: a coordinated balance of the cell cycle regulators cyclin D2 and p27Kip1. , 1998, Molecular endocrinology.

[27]  S. Korsmeyer,et al.  Bax-Deficient Mice with Lymphoid Hyperplasia and Male Germ Cell Death , 1995, Science.

[28]  S. Reed.  Cyclin E: in mid-cycle. , 1996, Biochimica et biophysica acta.

[29]  A. Storey,et al.  Granulosa cell survival and proliferation are altered in polycystic ovary syndrome. , 2008, The Journal of clinical endocrinology and metabolism.

[30]  T. Takezawa,et al.  Proliferative potential of endometrial stromal cells, and endometrial and placental expression of cyclin in the bovine. , 2003, The Journal of reproduction and development.

[31]  A. Hsueh,et al.  Hormonal Regulation of Apoptosis An Ovarian Perspective , 1997, Trends in Endocrinology & Metabolism.

[32]  N. Isobe,et al.  Localization of apoptotic cells in the cystic ovarian follicles of cows: a DNA-end labeling histochemical study. , 2000, Theriogenology.

[33]  R. Harman,et al.  Regulation of Fas Antigen (Fas, CD95)-Mediated Apoptosis of Bovine Granulosa Cells by Serum and Growth Factors1 , 2000, Biology of reproduction.

[34]  E. Gimeno,et al.  Intraovarian Localization of Growth Factors in Induced Cystic Ovaries in Rats , 2007, Anatomia, histologia, embryologia.

[35]  R. Cowan,et al.  Expression and Function of Fas Antigen Vary in Bovine Granulosa and Theca Cells During Ovarian Follicular Development and Atresia1 , 2000, Biology of reproduction.

[36]  C. Wilde,et al.  Functional expression of bcl-2 protein family and AIF in bovine mammary tissue in early lactation. , 2004, Journal of Dairy Research.

[37]  T. Nakao,et al.  The role of LH pulse frequency in ACTH-induced ovarian follicular cysts in heifers. , 2000, Animal reproduction science.

[38]  A. Bittles,et al.  Fas and Fas ligand messenger ribonucleic acid and protein expression in the rat corpus luteum during apoptosis-mediated luteolysis. , 1999, Biology of reproduction.

[39]  K. Teerds,et al.  Estrous cycle dependent changes in expression and distribution of Fas, Fas ligand, Bcl-2, Bax, and pro- and active caspase-3 in the rat ovary. , 2006, The Journal of endocrinology.

[40]  H. Billig,et al.  Survival factors regulating ovarian apoptosis -- dependence on follicle differentiation. , 2002, Reproduction.

[41]  P. Krammer.  CD95(APO-1/Fas)-mediated apoptosis: live and let die. , 1999, Advances in immunology.

[42]  E. Anderson,et al.  The polycystic ovarian (PCO) condition: apoptosis and epithelialization of the ovarian antral follicles are aspects of cystogenesis in the dehydroepiandrosterone (DHEA)-treated rat model. , 1997, Tissue & cell.

[43]  N. Isobe,et al.  Deficient proliferation and apoptosis in the granulosa and theca interna cells of the bovine cystic follicle. , 2007, The Journal of reproduction and development.

[44]  A. de Kruif,et al.  Aetiology and pathogenesis of cystic ovarian follicles in dairy cattle: a review. , 2006, Reproduction, nutrition, development.

[45]  B. Huppertz,et al.  The apoptosis cascade — morphological and immunohistochemical methods for its visualization , 1999, Anatomy and Embryology.

[46]  R. Harman,et al.  Ovarian follicular growth and atresia: the relationship between cell proliferation and survival. , 2004, Journal of animal science.

[47]  R. Bernards,et al.  CDK-Independent Activation of Estrogen Receptor by Cyclin D1 , 1997, Cell.

[48]  H. Ortega,et al.  Oestradiol induced inhibition of neuroendocrine marker expression in Leydig cells of adult rats. , 2006, Reproduction in domestic animals = Zuchthygiene.

[49]  E. Gimeno,et al.  Reproductive Biology and Endocrinology Open Access an Imbalance between Apoptosis and Proliferation Contributes to Follicular Persistence in Polycystic Ovaries in Rats , 2022 .

[50]  I. Dardick,et al.  Characterization of cytoskeletal proteins in basal cells of human parotid salivary gland ducts , 2004, Virchows Archiv A.

[51]  Ewert Bengtsson,et al.  Automatic Quantification of Immunohistochemically Stained Cell Nuclei Using Unsupervised Image Analysis , 1998, Analytical cellular pathology : the journal of the European Society for Analytical Cellular Pathology.

[52]  K. Okuda,et al.  Expressions of estrogen receptors in the bovine corpus luteum: cyclic changes and effects of prostaglandin F2alpha and cytokines. , 2007, The Journal of reproduction and development.

[53]  M. Skinner,et al.  Developmental and hormonal regulation of hepatocyte growth factor expression and action in the bovine ovarian follicle. , 1998, Biology of reproduction.

[54]  V. Padmanabhan,et al.  Developmental programming: prenatal androgen excess disrupts ovarian steroid receptor balance. , 2009, Reproduction.

[55]  E. Gimeno,et al.  Expression of Cytoskeletal Proteins in the Follicular Wall of Induced Ovarian Cysts , 2004, Cells Tissues Organs.

[56]  M. Peter,et al.  Two CD95 (APO‐1/Fas) signaling pathways , 1998, The EMBO journal.

[57]  J. Richards,et al.  Hormonal control of the cell cycle in ovarian cells: proliferation versus differentiation. , 1998, Biology of reproduction.

[58]  W. J. Silvia,et al.  Ovarian follicular cysts in dairy cows: an abnormality in folliculogenesis. , 2002, Domestic animal endocrinology.

[59]  W. Van den Broeck,et al.  Cell-specific localisation of apoptosis in the bovine ovary at different stages of the oestrous cycle. , 2006, Theriogenology.

[60]  M. Wiltbank,et al.  Follicular size and response to Ovsynch versus detection of estrus in anovular and ovular lactating dairy cows. , 2003, Journal of dairy science.