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在体外成熟的大鼠卵母细胞中,促黄体生成素加速溶酶体样细胞器的重新分布,先于核膜溶解。

Luteinizing hormone-accelerated redistribution of lysosome-like organelles preceding dissolution of the nuclear envelope in rat oocytes maturing in vitro.

作者信息

Ezzell R M, Szego C M

出版信息

J Cell Biol. 1979 Jul;82(1):264-77. doi: 10.1083/jcb.82.1.264.

Abstract

Maturation of the mammalian oocyte is characterized in part by dissolution of the nuclear envelope, or germinal vesicle breakdown (GVB). By fluorescence microscopy after vital uptake of acridine orange (AO), redistribution and perinuclear accumulation of organelles corresponding to lysosomes occur before GVB in rat oocytes undergoing meiotic maturation in vitro. In follicle-enclosed oocytes explanted during the preovulatory gonadotropin surge (GS) and individually cultured as such in chemically defined medium at approximately 22 degrees C, lysosomes aggregated into disperse clusters after 30 min; by 60 min, perinuclear concentration of lysosomes and their essential disappearance from the cortical ooplasm were observed. GVB occurred within 120 min. In contrast, follicle-enclosed oocytes explanted before the GS displayed a generally homogeneous distribution of lysosomes and intact GV for up to 5 h in culture. In oocytes aspirated from follicles before the GS, partially denuded of granulosa cells, and cultivated without added hormone, most lysosomes concentrated around the GV within 60 min, with GVB occurring generally by 120 min. Luteinizing hormone (LH) added in vitro to the isolated preparation at 3 or 30 x 10(-8) M sharply accelerated these events. The effects of LH, not seen with 1.5 x 10(-8) M hormone, were blocked by anti-LH IgG. Up to 60 x 10(-8) M follicle-stimulating hormone or 80 x 10(-8) M prolactin were ineffective in accelerating lysosome redistribution or GVB. After GVB, lysosomes became once again uniformly dispersed and unresponsive, even to 60 x 10(-8) M added LH, a finding consistent with tachyphylaxis of target cells by independent criteria. The present data, all statistically significant at P less than 0.05, demonstrate that mobilization of lysosomes before GVB is a specific response to factors that promote resumption of meiotic maturation of rat oocytes.

摘要

哺乳动物卵母细胞的成熟部分特征在于核膜溶解,即生发泡破裂(GVB)。在体外进行减数分裂成熟的大鼠卵母细胞中,通过吖啶橙(AO)的活体摄取后的荧光显微镜观察,对应于溶酶体的细胞器在GVB之前发生重新分布和核周积累。在排卵前促性腺激素高峰(GS)期间取出并在约22℃的化学限定培养基中单独培养的卵泡内卵母细胞,30分钟后溶酶体聚集形成分散的簇;到60分钟时,观察到溶酶体在核周集中并从皮质卵质中基本消失。GVB在120分钟内发生。相比之下,在GS之前取出的卵泡内卵母细胞在培养长达5小时内溶酶体通常呈均匀分布且生发泡完整。在GS之前从卵泡中吸出、部分去除颗粒细胞并在不添加激素的情况下培养的卵母细胞,大多数溶酶体在60分钟内集中在生发泡周围,GVB通常在120分钟时发生。体外向分离制剂中添加3或30×10⁻⁸ M的促黄体生成素(LH)可显著加速这些事件。1.5×10⁻⁸ M激素未观察到LH的作用,抗LH IgG可阻断其作用。高达60×10⁻⁸ M的促卵泡激素或80×10⁻⁸ M的催乳素在加速溶酶体重分布或GVB方面无效。GVB后,溶酶体再次均匀分散且无反应,即使添加60×10⁻⁸ M的LH也是如此,这一发现与通过独立标准确定的靶细胞快速脱敏一致。目前的数据在P小于0.05时均具有统计学意义,表明GVB之前溶酶体的动员是对促进大鼠卵母细胞减数分裂成熟恢复的因子的特异性反应。

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UPTAKE OF DYES AND DRUGS BY LIVING CELLS IN CULTURE.培养的活细胞对染料和药物的摄取
Life Sci (1962). 1964 Dec;3:1407-14. doi: 10.1016/0024-3205(64)90082-7.
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An electron microscope study of the rat ovum.大鼠卵子的电子显微镜研究。
J Biophys Biochem Cytol. 1959 Mar 25;5(2):327-42. doi: 10.1083/jcb.5.2.327.
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Functions of lysosomes.溶酶体的功能。
Annu Rev Physiol. 1966;28:435-92. doi: 10.1146/annurev.ph.28.030166.002251.

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