FSH and eCG impact follicles development and expression of ovarian FSHR and caspase-9 in mice

Document Type: Full paper (Original article)


1 Department of Reproduction Endocrinology, Life Science and Engineering College, Northwest University for Nationalities, Lanzhou 730030, Gansu Province, China

2 Affiliated Hospital, Northwest University for Nationalities, Lanzhou 730030, Gansu Province, China

3 Province Center for Prevention and Control of Animal Disease, Lanzhou 730046, Gansu Province, China

4 Engineering & Technology Research Center of Animal Cells of Gansu Province, Northwest University for Nationalities, Lanzhou 730030, Gansu Province, China


The study aimed to investigate the effects of FSH and eCG on the ovarian and follicular development, expression levels of FSHR and caspase-9 of ovaries in vivo. One hundred and five prepuberty mice were allocated into FSH-1, FSH-2, FSH-3, eCG-1, eCG-2, eCG-3 groups and control group (CG). Mice in FSH-1, FSH-2 and FSH-3 were intramuscularly injected with 5, 10 and 20 IU FSH twice (on day 0 and 4), respectively. Mice in eCG-1, eCG-2 and eCG-3 were intraperitoneally injected with 10, 20 and 40 IU eCG on day 0 and 4. Mice in the CG were injected with 0.5 ml normal saline on day 0 and 4. Left and right ovaries of each mouse were dissected aseptically on days 7, 14 and 21, respectively. The results showed that on days 14 and 21 the ovarian sizes and follicle numbers of FSH-3 and eCG-3 groups were greater than CG (P<0.05). FSHR mRNA of FSH-2 and eCG-1 were higher than CG on days 14 and 21 (P<0.05). FSHR proteins of FSH-3 were higher than CG on days 14 and 21 (P<0.05). Caspase-9 mRNA in FSH and eCG groups was less than CG. There were positive correlations between follicle numbers and FSH and eCG doses. FSHR protein expressions had positive correlations between ovarian weights and sizes of ovary and follicle numbers (r=0.971, P<0.05) in FSH-treated mice. Serum FSH concentrations of FSH-2, FSH-3, eCG-2 and eCG-3 groups were greater than that of CG. In conclusion, eCG and FSH promoted the ovarian development, follicle genesis, FSH secretion, FSHR mRNA and protein expressions in ovaries of mice. FSH and eCG inhibited the expression of ovarian caspase-9 mRNA.


Alejandro, CI; Manuel, XCV; Gustavo, RLC; Román, EC; Alejandro, CJC; Maximino, MM; Rubén, HC; Abel, VM; de Lourdes, JM; Pedro, SA and Eulogio, GL (2012). Effect of cloprostenol and fluorogestone acetate more PMSG on synchronization and no return to estrus in seasonal anestrus Dorper sheep. J. App. Sci. Res., 8: 1612-1614.

Ali, MS; Khandoker, MAM; Afroz, MA and Bhuiyan, AKFH (2012). Ovarian response to different dose levels of follicle stimulating hormone (FSH) in different genotypes of Bangladeshi cattle. Asian-Australas. J. Anim. Sci., 25: 52-58.

Asahara, S; Sato, A; Aljonaid, AA and Maruo, T (2003). Thyroid hormone synergizes with follicle stimulating hormone to inhibit apoptosis in porcine granulosa cells selectively from small follicles. Kobe J. Med. Sci., 49: 107-116.

Cao, J; Zhou, G; Liu, Y; Liao, G; Zhang, Q; Ye, K; Pan, D and Ou, C (2014). Activation of caspase-9 and its influencing factors in beef during conditioning. Animals. 8: 504-509.

Crawford, JL; Heath, DA; Haydon, LJ; Thomson, BP and Eckery, DC (2009). Gene expression and secretion of LH and FSH in relation to gene expression of GnRH receptors in the brushtail possum (Trichosurus vulpecula) demons-trates highly conserved mechanisms. Reproduction. 137: 129-140.

David, EN; Timothy, JP and Gary, CWE (2009). Veterinary reproduction and obstetrics. 9th Edn., Philadelphia, USA, Elsevier Medicine Press. PP: 127-189.

El-Nefiawy Nagwa, E (2011). Effect of exogenous prostag-landin E2 administration on ovarian follicle growth and angiogenesis in rat with reference to pregnant mare serum gonadotrophin. Egypt. J. Histol., 34: 251-259.

Ene, AC; Park, S; Edelmann, W and Taketo, T (2013). Caspase 9 is constitutively activated in mouse oocytes and plays a key role in oocyte elimination during meiotic prophase progression. Dev. Biol., 377: 213-223.

Erickson, GF and Shimasaki, S (2001). The physiology of folliculogenesis: the role of novel growth factors. Fertil. Steril., 76: 943-949.

Hillier, SG (2001). Gonadotropic control of ovarian follicular growth and development. Mol. Cell Endocrinol., 179: 39-46.

Hu, P; Huang, H; Liu, Y; Zhang, J; Lang, H and Zhang, G (2005). Effect of PMSG on the ovarian and uterine Kunming mouse immature and histology. Heilongjiang J. Anim. Reprod. China. 13: 6-7.

Hunzicker-Dunn, M and Maizels, ET (2006). FSH signaling pathways in immature granulosa cells that regulate target gene expression: branching out from protein kinase A. Cell Signal. 18: 1351-1359.

Layman, LC and McDonough, PG (2000). Mutations of follicle stimulating hormone-β and its receptor in human and mouse: genotype/phenotype. Mol. Cell Endocrinol., 161: 9-17.

Li, P; Yue, W; Pang, Y; Yu, X; Huang, Y; Ren, Y and Lv, L (2013). Effects of FSH and insulin on sheep ovarian follicular granulose cells in vitro culture. Acta Vet. et Zoot. Sin., 44: 1386-1391.

Maritza, PM; Jorg, G; Hermanan, MB; Claudia, G; Eberhard, N and Manuela, S (2000). Ovarian response to follicle-stimulating hormone (FSH) stimulation depends on the FSH receptor genotype. J. Clin. Endocrinol. Metabol., 85: 3365-3369.

McStay, GP; Salvesen, GS and Green, DR (2008). Over-
lapping cleavage motif selectivity of caspases: implications for analysis of apoptotic pathways. Cell Death Differ., 15: 322-331.

Menon, KMJ; Anil, K; Nair, LW and Helle, P (2007). Regulation of luteinizing hormone receptor mRNA expression by a specific RNA binding protein in the ovary. Mol. Cell Endocrinol., 260: 109-116.

Miro, F and Hillier, SG (1996). Modulation of granulosa cell deoxyribonucleic acid synthesis and differentiation by activin. Endocrinology. 137: 464-468.

Roberta, NC; Ana, BG; Duarte, G; Rodrigues, Q; Juliana, JHC; Gerlane, MS and Claudio, AP (2012). Effects of insulin and follicle-simulating hormone (FSH) during in vitro development of ovarian: goat preantral follicles and the relative mRNA expression for insulin and FSH receptors and cytochrome P450 aromatase in cultured follicles. Biol. Reprod., 87: 321-329.

Roy, SK and Albee, L (2000). Requirement for follicle-stimulating hormone action in the formation of primordial follicles during perinatal ovarian development in the hamster. Endocrinology. 114: 4449-4456.

Seekallu, SV; Toosi, BM; Duggavathi, R; Barrett, DMW; Davies, KL; Waldner, C and Rawlings, NC (2010). Ovarian antral follicular dynamics in sheep revisited: comparison among estrous cycles with three or four follicular waves. Theriogenology. 73: 670-680.

Susan, MJ and Roy, S (2000). Effects of exogenous FSH on follicular recruitment in a viviparous lizard Niveoscincus metallicus (Scincidae). Comparative biochemistry and physiology. Part A: Mol. Integrat. Physiol., 127: 487-493.

Thomas, FH and Vanderhyden, BC (2003). Oocyte granulosa cell interactions during mouse follicular development: regulation of kit ligand expression and its role in oocyte growth. Reprod. Biol. Endocrinol., 1: 1-7.

Wei, S; Chen, S; Gong, Z; Ouyang, X; An, L; Xie, K; Dong, J and Wei, M (2013a). Alarelin active immunization influences expression levels of GnRHR, FSHR and LHR proteins in the ovary and enhances follicular development in ewes. Anim. Sci. J., 84: 466-475.

Wei, S; Gong, Z; Ouyang, X; Xie, K and Wei, M (2013b). Modulation of expression, localization of FSHR and uterine development by GnRH agonist active immunization in ewes. J. Anim. Plant Sci., 19: 2888-2900.

Yuan, L; Wang, B; Piao, S; Tan, J and An, T (2008). Effect of PMSG on development of reproductive organs in different day-old immature mice. Acta Lab. Anim. Sci. Sin., 16: 338-341.

Zeleznik, A (2004). The physiology of follicle selection. Reprod. Biol. Endocrinol., 2: 31-34.

Zhang, C; Liu, L; Di, M; Zhao, D; Zhao, L; Xu, P and Rui, R (2007). Effects of the PMSG dosages on superovulation and oocyte quality in immature rats. Chinese J. Comp. Med., 17: 338-344. (in Chinese)

Zhang, C; Xia, G and Tsang, BK (2011). Interactions of thyroid hormone and FSH in the regulation of rat granulosa cell apoptosis. Front Biosci., 3: 1401-1413.

Zhou, J (1995). Research on estrus synchronization and superovulation in KM mice. Chinese J. Exp. Anim. Sci., 5: 139-141.

Zhou, XL; Teng, Y; Cao, R; Fu, H; Xiong, K; Sun, WX; Zhu, CC; Huang, XJ; Xiao, P and Liu, HL (2013). Rescue from dominant follicle atresia by follicle-stimulating hormone in mice. Genet. Mol. Res., 12: 2945-2952.