畜牧兽医学报 ›› 2013, Vol. 44 ›› Issue (7): 1043-1050.doi: 10.11843/j.issn.0366-6964.2013.07.008

• 遗传繁育 • 上一篇    下一篇

山羊卵巢间质细胞对小腔卵泡发育的影响

邱明宁,韩成全,杨忠财,刘军,权富生*,张涌*   

  1. (西北农林科技大学动物医学院,农业部动物生物技术重点开放实验室,杨凌 712100)
  • 收稿日期:2013-01-11 出版日期:2013-07-23 发布日期:2013-07-23
  • 通讯作者: 权富生,博士,副教授,主要从事哺乳动物胚胎工程及发育生物学方面的教学和研究,E-mail: quanfusheng@nwsuaf.edu.cn;张涌,博士,教授,从事动物胚胎工程及发育生物学方面的研究,E-mail: zhangyong@nwsuaf.edu.cn
  • 作者简介:邱明宁(1986-),男,辽宁铁岭人,硕士生,主要从事发育生物学、动物克隆与转基因技术研究,E-mail: lcuzfy@163.com
  • 基金资助:

    “十二五”农村领域国家科技计划课题;“863”项目 (2011AA100303)

Effect of Ovarian Stromal Cells on Co-cultured Small Antral Follicles in the Goat

QIU Ming-ning, HAN Cheng-quan, YANG Zhong-cai, LIU Jun, QUAN Fu-sheng*, ZHANG Yong*   

  1. (Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China
  • Received:2013-01-11 Online:2013-07-23 Published:2013-07-23

摘要:

为了能有效利用山羊小腔卵泡和研究相关小腔卵泡发育的机理,将山羊小腔卵泡与卵巢皮质间质细胞、卵泡膜细胞和卵巢髓质间质细胞体外共培养,测定培养第5天卵泡直径、培养液中类固醇激素分泌量、卵母细胞特异性分泌因子gdf9bmp15基因的表达和卵泡颗粒细胞以及卵母细胞凋亡与抗凋亡基因表达。研究结果显示,山羊小腔卵泡与卵巢间质细胞共培养5 d,卵泡直径增加明显大于小腔卵泡单独培养组(P<0.05);小腔卵泡与卵巢皮质间质细胞共培养5 d后,小腔卵泡分泌的雌二醇激素的量高于小腔卵泡单独培养组(P<0.01),为413.95 pg·mL-1;小腔卵泡与卵巢髓质间质细胞共培养5 d后,小腔卵泡分泌孕酮的量(11.695 ng·mL-1)极显著高于小腔卵泡单独培养组和其他共培养组(P<0.01),小腔卵泡与卵泡膜细胞共培养5 d后,小腔卵泡分泌的孕酮的量高于小腔卵泡单独培养组(P<0.05);小腔卵泡与卵巢皮质间质细胞、髓质间质细胞以及与卵泡膜细胞共培养5 d后,其卵母细胞特异性分泌因子gdf9基因和bmp15基因表达显著上调(P<0.01);与卵巢间质细胞共培养5 d后,小腔卵泡内卵丘细胞和卵母细胞抗凋亡基因表达上调(P<0.05),小腔卵泡内卵丘细胞和卵母细胞抗凋亡基因表达下调 (P<0.05)。表明卵巢间质细胞促进小腔卵泡的生长以及类固醇激素如雌二醇和孕酮分泌,促进小腔卵泡内卵母细胞特异性分泌的生长因子(gdf9bmp15)基因的表达,同时抑制小腔卵泡的凋亡。尤其是卵巢卵泡膜细胞与小腔卵泡共培养后明显提高小腔卵泡的抗凋亡能力。

Abstract:

To investigate the mechanism of follicular development in the goat, our present study were designed to co-culture small antral follicles with ovarian cortex stromal cells, theca cells and ovarian medullary stromal cells in vitro for 5 d, respectively, and then to measure the change of diameter of follicles, steroid hormone productions, expressions of oocyte secreted factors (gdf9 and bmp15) and inhibitory apoptotic and pro-apoptotic genes. The results showed that the diameter of follicles were increased significantly in co-cultured groups than which cultured alone after 5 d culture (P<0.05); estradiol production increased in small antral follicles (413.95 pg·mL-1) after co-cultured with ovarian cortical stromal cells for 5 d (P<0.01); progesterone production extremely increased in small antral follicles (11.695 ng·mL-1) when co-cultured with ovarian medullary stromal cells for 5 d (P<0.01), progesterone production increased in small antral follicles when co-cultured with theca cells for 5 d (P<0.05); expressions of gdf9 and bmp15 mRNA in the oocytes in follicles co-cultured with ovarian stromal cells and theca cells were increased (P<0.01); expressions of inhibitory apoptotic genes in cumulus cells and oocytes in small antral follicles co-cultured increased (P<0.05), and expressions of pro-apoptotic genes in cumulus cells and oocytes in small antral follicles co-cultured decreased (P<0.05). These results indicated that ovarian stromal cells and theca cells could promote growing of small antral follicles, steroid hormone productions, and expressions of oocyte secreted factors (gdf9 and bmp15), and inhibit apoptosis of small antral follicles; especially enhance the capacity of small antral follicles significantly through inhibiting apoptosis after co-cultured with theca cells.

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