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通过原位杂交检测卤虫发育过程中钠钾ATP酶α2和β信使核糖核酸的差异表达。

Differential expression of the alpha 2 and beta messenger RNAs of Na,K-ATPase in developing brine shrimp as measured by in situ hybridization.

作者信息

Sun D Y, Guo J Z, Hartmann H A, Uno H, Hokin L E

机构信息

Department of Pathology, University of Wisconsin, Madison 53706.

出版信息

J Histochem Cytochem. 1992 Apr;40(4):555-62. doi: 10.1177/40.4.1313064.

Abstract

We used in situ hybridization histochemistry with synthetic oligonucleotide probes to localize the mRNAs encoding the alpha 2- and beta-mRNAs of Na,K-ATPase during development of the brine shrimp Artemia. The mRNAs of the alpha 2- and beta-subunit were of low abundance in the cysts; in addition, less mRNA of the beta-subunit was localized. During emergence (12 hr), there was an increase in alpha 2-subunit mRNA in the gut mucosa, but there was a burst in beta-subunit mRNA throughout. As development progressed, the mRNAs of both the alpha 2- and beta-subunits showed a distinct pattern of expression in which the mRNA in the salt gland was of greatest abundance, followed by epidermal cells and gut mucosa. After 36 hr the alpha 2-subunit mRNA began to decrease in all positive cells but still remained highest in the salt gland and the brain region, while the mRNA of the beta-subunit kept increasing in the gut mucosa. Finally, the greatest abundance of the beta-subunit mRNA shifted from the salt gland to the antenna gland and the epidermal cells in the tail region, but the alpha 2-subunit mRNA did not. The more widespread distribution of the beta-mRNA than alpha 2-mRNA at certain stages (e.g., there was no alpha 2-mRNA in the antenna gland at the adult stage) is in all likelihood due to the marked drop in the alpha 2-subunit and a rise in alpha 1-subunit previously seen by Peterson et al. on polyacrylamide gel electrophoresis, as development progresses.

摘要

我们使用合成寡核苷酸探针进行原位杂交组织化学,以在卤虫发育过程中定位编码钠钾ATP酶α2和β亚基mRNA。α2和β亚基的mRNA在囊肿中丰度较低;此外,β亚基的mRNA定位较少。在孵化期(12小时),肠道黏膜中α2亚基mRNA增加,但β亚基mRNA在全身都有激增。随着发育进行,α2和β亚基的mRNA均呈现出独特的表达模式,其中盐腺中的mRNA丰度最高,其次是表皮细胞和肠道黏膜。36小时后,α2亚基mRNA在所有阳性细胞中开始减少,但在盐腺和脑区仍保持最高水平,而β亚基mRNA在肠道黏膜中持续增加。最后,β亚基mRNA的最高丰度从盐腺转移到触角腺和尾部区域的表皮细胞,但α2亚基mRNA没有。在某些阶段(例如,成年期触角腺中没有α2 mRNA),β mRNA的分布比α2 mRNA更广泛,这很可能是由于随着发育进行,先前Peterson等人在聚丙烯酰胺凝胶电泳上观察到的α2亚基显著下降和α1亚基上升所致。

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