摘要
为了研究八眉猪卵泡闭锁过程中卵泡颗粒细胞的凋亡情况、卵泡组织结构变化以及卵泡膜内层毛细血管变化特点,试验在体视显微镜下观察并将卵泡分为健康卵泡、早期闭锁卵泡和晚期闭锁卵泡,用Hoechst 33258对闭锁不同时期卵泡颗粒细胞凋亡情况进行检测。采用苏木精-伊红(H.E.)染色法对卵泡膜层组织结构进行观察,CD34免疫组化染色对卵泡膜上毛细血管的分布进行观察。结果表明:随着卵泡闭锁程度的加深,卵泡颗粒细胞逐渐脱落进入卵泡腔,颗粒细胞凋亡增多,分布在卵泡膜内层的毛细血管随着卵泡闭锁程度的加深逐渐消失。说明卵泡颗粒细胞凋亡和卵泡膜上微血管消失是八眉猪卵泡闭锁的重要特征。
引文
[1] TOWNSON D H,COMBELLES C M H.Ovarian Follicular Atresia[M].London:In Tech,2012.
[2] HSUEH A J,BILLIG H,TSAFRIRI A,et al.Ovarian follicle atresia:a hormonally controlled apoptotic process[J].Endocr Rev,1994,15(6):707-724.
[3] SEN A,HAMMES S R.Granulosa cell-specific androgen receptors are critical regulators of ovarian development and function [J].Mol Endocrinol,2010,24(7):1393-1403.
[4] 张金璧,林飞,潘增祥,等.猪不同程度闭锁卵泡判定方法的比较研究[J].南京农业大学学报,2013,36(1):115-119.
[5] 林飞.猪卵泡闭锁的特征与miRNA调控研究[D].南京:南京农业大学,2011.
[6] JOHNSON A L,BRIDGHAM J T.Caspase-mediated apoptosis in the vertebrate ovary[J].Reproduction,2002,124(1):19-27.
[7] ZHANG J M,WANG H C,WANG H X,et al.Oxidative stress and activities of caspase-8,-9,and -3 are involved in cryopreservation-induced apoptosis in granulosa cells [J].Eur J Obstet Gynecol Reprod Biol,2013,166(1):52-55.
[8] CHO S W,LEE S H,CHUNG M K,et al.The relationship between the expression of the apoptosis-related genes (caspase-3,8,PARP) in granulosa cells and the outcome of IVF patients[J].Fertil Steril,2004,82(3):73.
[9] MATSUI T,MANABE N,GOTO Y,et al.Expression and activity of Apaf1 and caspase-9 in granulosa cells during follicular atresia in pig ovaries[J].Reproduction,2003,126(1):113-120.
[10] BOONE D L,TSANG B K.Caspase-3 in the rat ovary:localization and possible role in follicular atresia and luteal regression[J].Biol Reprod,1998,58(6):1533-1539.
[11] ZHU L,YUAN H,GUO C,et al.Zearalenone induces apoptosis and necrosis in porcine granulosa cells via a caspase-3- and caspase-9-dependent mitochondrial signaling pathway[J].J Cell Physiol,2012,227(5):1814-1820.
[12] FENWICK M A,HURST P R.Immunohistochemical localization of active caspase-3 in the mouse ovary:growth and atresia of small follicles[J].Reproduction,2002,124(5):659-665.
[13] MATIKAINEN T,PEREZ G I,ZHENG T S,et al.Caspase-3 gene knockout defines cell lineage specificity for programmed cell death signaling in the ovary[J].Endocrinology,2001,142(6):2468-2480.
[14] BERARDINELLI P,RUSSO V,MARTELLI A,et al.Colocalization of DNA fragmentation and Aaspase-3 activation during atresia in pig antral follicles[J].Anat Histol Embryol,2004,33(1):23-27.
[15] MANABE N,KIMURA Y,UCHIO K,et al.Regulatory mechanisms of granulosa cell apoptosis in ovarian follicle atresia [M].Berlin:Springer Netherlands,2002.
[16] MIYAKE Y,MATSUMOTO H,YOKOO M,et al.Expression and glycosylation with polylactosamine of CD44 antigen on macrophages during follicular atresia in pig ovaries[J].Biol Reprod,2006,74(3):501-510.
[17] MATTIOLI M,BARBONI B,TURRIANI M,et al.Follicle activation involves vascular endothelial growth factor production and increased blood vessel extension1[J].Biol Reprod,2001,65(4):1014-1019.
[18] FERANIL J B,ISOBE N,NAKAO T.Changes in the thecal vasculature during follicular atresia in the ovary of swamp buffalo[J].J Reprod Dev,2004,50(3):315-321.
[19] SATO E,KAWAHARA M,SAKURAI M.Follicular microvasculature in the porcine and bovine ovary visualized by SEM of vascular corrosion casts[J].Morphology,2008,4(3):1-6.
[20] JIANG J,MACCHIARELLI G,MIYABAYASHI K,et al.Follicular microvasculature in the porcine ovary [J].Cell Tissue Res,2002,310(1):93-101.
[21] SUZUKI T,SASANO H,TAKAYA R,et al.Cyclic changes of vasculature and vascular phenotypes in normal human ovaries[J].Hum Reprod,1998,13(4):953-959.