不同成熟培养时间对家猫卵母细胞皮质颗粒分布的影响

doi:10.7668/hbnxb.2011.03.020

摘要/Abstract

摘要： 对家猫卵母细胞皮质颗粒染色,以检测不同成熟培养时间对猫卵母细胞胞质成熟的影响.将成熟培养24,28,32,36,43 h的家猫卵母细胞,以3.7%的多聚甲醛固定,之后分别对各组卵母细胞进行FITC-LCA和PI染料染色,在激光共聚焦显微镜下观察皮质颗粒(CGs)在卵母细胞胞质中的分布.结果表明,不同培养时间的卵母细胞处于MⅡ期时,CGs分布于整个家猫卵母细胞中,并未出现无皮质颗粒区(CGFD);与其他各组相比,32 h组卵母细胞皮质层中的CGs更多.通过皮质颗粒染色证明32 h组卵母细胞胞质成熟效果最好,32 h是家猫卵母细胞体外成熟的最佳时间.

关键词: 家猫, 卵母细胞, 皮质颗粒

Abstract: In order to evaluate the effect of duration on oocyte cytoplasm maturation,the oocytes matured 24,28,32,36 and 43 h were fixed in 3.7% paraformaldehyde and stained in FITC-LCA and PI,and the distribution of cortical granules(CGs) was observed to evaluate the cytoplasmic maturation.CGs were found throughout the oocyte cytoplasm.The CGFD didn't appear in oocytes of domestic cat.There were more CGs located in the cortex of the oocytes which were matured 32 h than the other groups.CGs decreased in oocytes matured 36 h and 43 h.The resultes showed that oocytes matured 32 h got ready to be sitimulated,and after 32 h the oocytes were degraded.Therefore,32 h was the best maturation duration of domestic cat oocytes IVM.

Key words: Domestic cat, Oocyte, Cortical granules

中图分类号:

S88.293

王新庄, 秦佳晨, 任卫青, 何俊丹, 吕婧玉. 不同成熟培养时间对家猫卵母细胞皮质颗粒分布的影响[J]. 华北农学报, 2011, 26(3): 95-99. doi: 10.7668/hbnxb.2011.03.020.

WANG Xin-zhuang, QIN Jia-chen, REN Wei-qing, HE Jun-dan, LU Jing-yu. Effects of Duration on Oocyte Cytoplasm Maturation in vitro with CGs Staining[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2011, 26(3): 95-99. doi: 10.7668/hbnxb.2011.03.020.

http://www.hbnxb.net/CN/Y2011/V26/I3/95

参考文献

[1] Gulyas B J.Cortical granules of mammalian eggs[J].Int Rev Cytol,1980,63:357-392.

[2] Damiani P,Fissore R A,Cibelli J B,et al.Evaluation of developmental competence,nuclear and ooplasmic maturation of caff oocytes[J].Mol Reprod Dev,1996,45,521-534.

[3] 马红.激素对牛卵母细胞体外成熟的影响[J].华北农学报,2009,24(增刊):338-339.

[4] 刘海军,刘灵,刘玉堂,等.受体卵母细胞来源和供体细胞类型对山羊体细胞核移植融合率的影响[J].华北农学报,2008,23(1):41-44.

[5] 王星,罗光彬,刘永,等.生长激素和胰岛素对猪卵母细胞体外成熟及卵裂的影响[J].河南农业科学,2009(1):100-103.

[6] 李晓霞,曹平华,邓星,等.不同浓度的牛血清及牛血清白蛋白对水牛卵母细胞体外成熟的影响[J].河南农业科学,2008(7):108-111.

[7] Miyara F,Aubriot F X,Glissant A,et al.Multiparameter analysis of human oocytes at metaphase Ⅱ stage after IVF failure in non-male infertility[J].Hum Reprod,2003,18:1494-1503.

[8] Zamboni L.Ultrastructure of mammalian oocytes and ova[J].Biol Reprod,1970,2:44-63.

[9] Hoodbhoy T,Talbot P.Mammalian cortical granules:contents,fate,and function[J].Mol Reprod Dev,1994,39:439-448.

[10] Ducibella T,Buetow J.Competence to undergo normal,fertilizationinduced cortical activation develops after metaphase Ⅰ of meiosis in mouse oocytes[J].Der Biol,1994,165:95-104.

[11] Schuel H.Function of egg cortical granules[D].New York,USA:Metz CB and Monroy A,1985.

[12] Gwatkin R B L.Fertilization mechanisms in man and mammals[M].New York:Plenum,1997.

[13] Yanagimachi R.Mammalian fertilization[D].New York:Knobil E and Neill J D,1994.

[14] DucibellaT,Kurasawa S,Rangarajan S,et al.Precocious loss of cortical granules during mouse oocyte meiotic maturation and correlation with an egg-induced modification of the Zona pellucida[J].Dev Biol,1990,137:46-55.

[15] Ducibella T,Duffy P,Reindollar R,et al.Changes in the distribution of mouse oocyte cortical granules and ability to undergo the cortical reaction during gonadotropin-stimulated meiotic maturation and aging in vivo[J].Biol Reprod,1990,43:870-876.

[16] Liu M,Sims D,Calarco P,et al.Biochemical heterogeneity,migration,and pre-fertilization release of mouse oocyte cortical granules[J].Reprod Biol Endocrino,2003,1(1):77.

[17] Byers A P,Barone M A,Donoghue A M,et al.Matration domestic cat oocytes does not express a cortical granule-free domain[J].Biol Reprod,1992,47:709-715.

[18] Mikkelsen A L,Lindenbegrg S.Morphology of in vitro matured oocytes:Impact on fertility potential and embryo quality[J].Hum RePord,2001,16:1714-1718.

[19] Khatir H,Lonegran P,Caorlan C,et al.Prepubertal bovine oocyte:a negative model for studying oocyte development competence[J].Mol Reprod Dev,1996,45:231-239.

[20] Naito K,Fukuda Y,Toyoda Y.Effects of porcine follicular fluid on male pornucleus of maturation in porcine oocytes matured in vitor[J].Gamet Research,1988,21:289-295.

[21] 李荣凤,薛晓先,刘哲,等.牛卵母细胞皮层颗粒荧光染色法质成熟鉴定[J].畜牧兽医学报,2000,31(3):203-210.

[22] De Felici M,Siracusa G.Spontaneous hardening of the zona pellucida of mouse oocytes during in vitro culture[J].Gamete Res,1982,6:107-113.

[23] Zhang X,Rutledge J,Amrstrong D T.Studies on zona hardening in rat oocytes that are matured in vitor in a serum-free medium[J].Mol ReProd Dev,1991,28:292-296.

[24] Downs S M,Schoreder A C,Eppig J J.Seurm maintains the fertilizability of mouse oocytes matured in vitor by preventing the hardening of the Zona pellucida[J].Gamete Res,1986,15:115-122.

[25] Fujii Y,Yoshioka T,Saskai J.Fetal calf serum increased the Zona pellucida penetrability of rat oocytes matured in vitor[J].Acta Med Okayama,1990,44:203-208.

[26] Dell' Aquila M E,Cho Y S,Minoial P,et al.Effects of follicular fluid supplementation of in-vitro maturation medium on the fertilization and development of equine oocytes after in-vitor fertilization or intracytoplasmic sperm injection[J].Hum Reprod,1997,12:2766-2772.

[27] Peter K,Gregoyr S K,Riehadr M S.Modifications of the mouse Zona pellucida during oocyte muatration and egg activation:Effects of new born calf serum and fetuin[J].Biol Reprod,1991:783-787.

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