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| Morphological and Hormonal Identification of Porcine Atretic Follicles and Relationship Analysis of Hormone Receptor Levels During Granulosa Cell Apoptosis In vivo |
| YU De-bing, YU Min-li, LIN Fei, JIANG Bao-chun, YANG Li-na, WANG Si-yu, ZHAO Ying , WNAG Zheng-chao |
| Department of Animal Genetics, Breeding and Reproduction, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, P.R.China |
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摘要 Recent reports have demonstrated that follicular atresia is initiated or caused by granulosa cell apoptosis followed by theca cell degeneration in mammalian ovaries, but the mechanism of follicular atresia is still to be elucidated. Therefore, our present study was designed to examine our hypothesis that the changes of follicular microenvironment induce the granulosa cell apoptosis during pocrine follicular atresia in vivo. We firstly isolated intact porcine antral follicles and identified them into three groups, healthy follicles (HF), early atretic follicles (EAF) and progressed atretic follicles (PAF) through morphology and histology. To further confirm their status, we detected hormone levels in follicular fluids and the expression level of apoptosis gene Bax in granulosa cells. The rate of progesterone (P) and estradiol (E2) was increased with the expression of Bax, indicating hormone can be used as a marker of granulosa cell apoptosis or follicular atresia. Finally, we analyzed the expression level of hormone receptor genes in granulosa cells and their relationship with follicular atresia. In PAF, the expression of Progesterone receptor (PGR) was increased significantly while estradiol receptor (ER) had no notable changes, which suggesting the increased-PGR accelerated the effect of P-stimulated granulosa cell apoptosis. The dramatic increasing of androgen receptor (AR) expression in PAF and the obvious increase of tumor necrosis factor-α receptor (TNFR) in EAF indicated that there are different pathways regulating granulosa cell apoptosis during follicular atresia. Together, our results suggested that different pathways of granulosa cell apoptosis was induced by changing the follicular microenvironment during follicular atresia.
Abstract Recent reports have demonstrated that follicular atresia is initiated or caused by granulosa cell apoptosis followed by theca cell degeneration in mammalian ovaries, but the mechanism of follicular atresia is still to be elucidated. Therefore, our present study was designed to examine our hypothesis that the changes of follicular microenvironment induce the granulosa cell apoptosis during pocrine follicular atresia in vivo. We firstly isolated intact porcine antral follicles and identified them into three groups, healthy follicles (HF), early atretic follicles (EAF) and progressed atretic follicles (PAF) through morphology and histology. To further confirm their status, we detected hormone levels in follicular fluids and the expression level of apoptosis gene Bax in granulosa cells. The rate of progesterone (P) and estradiol (E2) was increased with the expression of Bax, indicating hormone can be used as a marker of granulosa cell apoptosis or follicular atresia. Finally, we analyzed the expression level of hormone receptor genes in granulosa cells and their relationship with follicular atresia. In PAF, the expression of Progesterone receptor (PGR) was increased significantly while estradiol receptor (ER) had no notable changes, which suggesting the increased-PGR accelerated the effect of P-stimulated granulosa cell apoptosis. The dramatic increasing of androgen receptor (AR) expression in PAF and the obvious increase of tumor necrosis factor-α receptor (TNFR) in EAF indicated that there are different pathways regulating granulosa cell apoptosis during follicular atresia. Together, our results suggested that different pathways of granulosa cell apoptosis was induced by changing the follicular microenvironment during follicular atresia.
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Received: 02 November 2012
Accepted:
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| Fund: This study was supported by the National Basic Research Program of China (2007CB947403), the National Natural Science Foundation of China (31101705) and the grant from Students Practice and Innovation Training Program of Jiangsu Province, China (JSS0909). |
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Corresponding Authors:
YU De-bing, Tel/Fax: +86-25-84395036, E-mail: yudb79@163.com
E-mail: yudb79@163.com
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| About author: YU De-bing, Tel/Fax: +86-25-84395036, E-mail: yudb79@163.com |
Cite this article:
YU De-bing, YU Min-li, LIN Fei, JIANG Bao-chun, YANG Li-na, WANG Si-yu, ZHAO Ying , WNAG Zheng-chao.
2014.
Morphological and Hormonal Identification of Porcine Atretic Follicles and Relationship Analysis of Hormone Receptor Levels During Granulosa Cell Apoptosis In vivo. Journal of Integrative Agriculture, 13(5): 1058-1064.
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| Alonso-Pozos I, Rosales-Torres A M, Avalos-Rodriguez A, Vergara-Onofre M, Rosado-Garcia A. 2003. Mechanism of granulosa cell death during follicular atresia depends on follicular size. Theriogenology, 60, 1071-1081 Basini G, Mainardi G L, Bussolati S, Tamanini C. 2002. Steroidogenesis, proliferation and apoptosis in bovine granulosa cells, role of tumour necrosis factor-alpha and its possible signalling mechanisms. Reproduction, Fertility and Development, 14, 141-150 Chen Y, Jefferson W N, Newbold R R, Padilla-Banks E, Pepling M E. 2007. Estradiol, progesterone, and genistein inhibit oocyte nest breakdown and primordial follicle assembly in the neonatal mouse ovary in vitro and in vivo. Endocrinology, 148, 3580-3590 Guthrie H D, Grimes R W, Cooper B S, Hammond J M. 1995. Follicular atresia in pigs, measurement and physiology. Journal of Animal Science, 73, 2834-2844 Hussein M R. 2005. Apoptosis in the ovary, molecular mechanisms. Human Reproduction Update, 11, 162-177 Inoue N, Matsuda-Minehata F, Goto Y, Sakamaki K, Manabe N. 2007. Molecular characteristics of porcine Fas-associated death domain (FADD) and procaspase-8 Journal of Reproduction and Development, 53, 427-436.Jiang J Y, Cheung C K, Wang Y, Tsang B K. 2003. Regulation of cell death and cell survival gene expression during ovarian follicular development and atresia. Frontiers in Bioscience, 8, d222-d237. Jolly P D, Smith P R, Heath D A, Hudson N L, Lun S, Still L A, Watts C H, McNatty K P. 1997. Morphological evidence of apoptosis and the prevalence of apoptotic versus mitotic cells in the membrana granulosa of ovarian follicles during spontaneous and induced atresia in ewes. Biology of Reproduction, 56, 837-846Kayampilly P P, Wanamaker B L, Stewart J A, Wagner C L, Menon K M. 2010. Stimulatory effect of Insulin on 5alpha-reductase type 1SRD5A1 expression through an Akt-dependent pathway in ovarian granulosa cells. Endocrinology, 151, 5030-5037 Kaipia A, Hsueh A J. 1997. Regulation of ovarian follicle atresia. Annual Review of Physiology, 59, 349-363 Kezele P, Skinner M K. 2003. Regulation of ovarian primordial follicle assembly and development by estrogen and progesterone, endocrine model of follicle assembly. Endocrinology, 144, 3329-3337 Lin P F, Hao Y B, Guo H L, Liu H L, Rui R. 2010. Role of Fas/FasL on apoptosis of porcine follicular granulosa cells derived from isolated follicles during culture in vitro. Zoological Research, 31, 268-274 (in Chinese) Lin P, Rui R. 2010. Effects of follicular size and FSH on granulosa cell apoptosis and atresia in porcine antral follicles. Molecular Reproduction and Development, 77, 670-678 Liou C M, Yang A L, Kuo C H, Tin H, Huang C Y, Lee S D. 2010. Effects of 17beta-estradiol on cardiac apoptosis in ovariectomized rats. Cell Biochemistry and Function, 28, 521-528 Manabe N, Goto Y, Matsuda-Minehata F, Inoue N, Maeda A, Sakamaki K, Miyano T. 2004. Regulation mechanism of selective atresia in porcine follicles, regulation of granulosa cell apoptosis during atresia. Journal of Reproduction and Development, 50, 493-514 Manabe N, Imai Y, Ohno H, Takahagi Y, Sugimoto M, Miyamoto H. 1996. Apoptosis occurs in granulosa cells but not cumulus cells in the atretic antral follicles in pig ovaries. Experientia, 52, 647-651 Manabe N, Myoumoto A, Tajima C, Fukumoto M, Nakayama M, Uchio K, Yamaguchi M, Miyamoto H. 2000. Immunochemical characteristics of a novel cell death receptor and a decoy receptor on granulosa cells of porcine ovarian follicles. Cytotechnology, 33, 189-201 Matsuda-Minehata F, Goto Y, Inoue N, Sakamaki K, Chedrese P J, Manabe N. 2007. Anti-apoptotic activity of porcine cFLIP in ovarian granulosa cell lines. Molecular Reproduction and Development, 74, 1165- 1170. Matsuda-Minehata F, Inoue N, Goto Y, Manabe N. 2006. The regulation of ovarian granulosa cell death by pro- and anti-apoptotic molecules. Journal of Reproduction and Development, 52, 695-705 Moor R M, Dai Y, Lee C, Fulka J. 1998. Oocyte maturation and embryonic failure. Human Reproduction Update, 4, 223-236 Moosavifar N, Mohammadpour A H, Jallali M, Karimiz G, Saberi H. 2010. Evaluation of effect of silymarin on granulosa cell apoptosis and follicular development in patients undergoing in vitro fertilization. Eastern Mediterranean Health Journal, 16, 642-645 Mussche S, D’Herde K. 2001. Contribution of progesterone, follicle stimulating hormone and glucocorticoids in survival of serum-free cultured granulosa cell explants. Journal of Endocrinology, 169, 321-331 Sugimoto M, Kagawa N, Morita M, Kume S, Wongpanit K, Jin H, Manabe N. 2010. Changes in the expression of decoy receptor 3 in granulosa cells during follicular atresia in porcine ovaries. Journal of Reproduction and Development, 56, 467-474 Tabarowski Z, Szoltys M, Bik M, Slomczynska M. 2005. Atresia of large ovarian follicles of the rat. Folia Histochemica et Cytobiologica, 43, 43-50 Wang W,Wang L, Li X X, Chen X, Zhang H Y, He Y, Wang J J, Zhao Y Y, Zhang B L, Xu Y X. 2010. Effect of interrupted endogenous BMP/Smad signaling on growth and steroidogenesis of porcine granulosa cells. Journal of Zheijang University (Science B), 11, 719-727 Zhang X, Tong N, Li Y. 2010a. Concurrently using rosiglitazone prevents glucosamine-induced islet beta- cell apoptosis and dysfunction. Cell Biochemistry and Function, 28, 509-514 Zhang X, Zhang L, Huo S, Wang J, Cui S. 2010b. Neonatal superior ovarian nerve transection inhibits follicle development by enhancing follicular atresia and suppressing granulosa cell proliferation in rats. Reproduction, Fertility, and Development, 22, 1148- 1158. |
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