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卵因子对海胆精子中环核苷酸代谢的影响。

Effects of egg factors on cyclic nucleotide metabolism in sea urchin sperm.

作者信息

Garbers D L, Hardman J G

出版信息

J Cyclic Nucleotide Res. 1976;2(2):59-70.

PMID:4475
Abstract

Cyclic AMP in Strongylocentrotus purpuratus sperm was elevated approximately 2-fold by theophylline or 1-methyl-3-isobutylxanthine. Factors released from sea urchin eggs (FRE) elevated sperm cyclic AMP by about 7-fold within 1 min, and the combination of FRE with theophylline increased sperm cyclic AMP up to 100-fold within 1 min. Cyclic GMP in sea urchin sperm was slightly elevated by theophylline, but was lowered by FRE. Cyclic GMP in sperm treated with FRE plus theophylline was not higher than in sperm treated with theophylline alone. The ability of FRE-containing sea water to increase sperm cyclic AMP in the presence of theophylline was altered only slightly if at all by boiling, but it was decreased by about 50% by dialysis and destroyed by ashing. Filtration of FRE on Sephadex G-50 columns yielded two peaks of cyclic AMP-elevating activity. One peak (peak I) was eluted at the column void volume, and the other (peak II) was retained by the column. The cyclic GMP-lowering activity was located in fractions approximately corresponding to peak I of cyclic AMP-elevating activity. Dialysis of FRE-containing sea water before its application to the G-50 column virtually eliminated peak II of the cyclic AMP-elevating activity. When the cyclic AMP-elevating activity in peak I was filtered on Bio Gel A-5m columns, it also migrated at or near the column void volume. Fractions corresponding to peak I contained material that inhibited both guanylate and adenylate cyclase activities in broken cell preparations of sperm and guanylate cyclase from rat lung. The inhibitory material was stable to boiling, non-dialyzable, and destroyed by ashing. Under a variety of conditions, FRE-containing sea water or cyclic AMP-elevating peaks I or II did not stimulate sperm adenylate cyclase activity in broken cell preparations.

摘要

在紫海胆精子中,茶碱或1-甲基-3-异丁基黄嘌呤可使环磷酸腺苷(cAMP)升高约2倍。海胆卵释放因子(FRE)在1分钟内可使精子cAMP升高约7倍,而FRE与茶碱联合使用可在1分钟内使精子cAMP升高达100倍。茶碱可使海胆精子中的环磷酸鸟苷(cGMP)略有升高,但FRE可使其降低。用FRE加茶碱处理的精子中的cGMP并不高于仅用茶碱处理的精子。含FRE的海水在茶碱存在下增加精子cAMP的能力,即使经过煮沸也仅有轻微改变,但若有改变的话;但经透析后降低约50%,经灰化则被破坏。将FRE在葡聚糖G-50柱上过滤,得到两个升高cAMP活性的峰。一个峰(峰I)在柱的空体积处被洗脱,另一个峰(峰II)被柱保留。降低cGMP的活性位于大致对应于升高cAMP活性峰I的级分中。在将含FRE的海水应用于G-50柱之前进行透析,实际上消除了升高cAMP活性的峰II。当峰I中的升高cAMP活性在生物凝胶A-5m柱上过滤时,它也在柱的空体积处或其附近迁移。对应于峰I的级分含有抑制精子破碎细胞制剂中的鸟苷酸环化酶和腺苷酸环化酶活性以及大鼠肺鸟苷酸环化酶的物质。该抑制物质对煮沸稳定,不可透析,经灰化被破坏。在各种条件下,含FRE的海水或升高cAMP的峰I或峰II在精子破碎细胞制剂中均不刺激精子腺苷酸环化酶活性。

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