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激素受体定位的放射自显影和生化研究。

Autoradiographic and biochemical studies of hormone receptor localization.

作者信息

Sheridan P J, Buchanan J M, Anselmo V

出版信息

J Histochem Cytochem. 1981 Jan;29(1A Suppl):195-200.

PMID:7288155
Abstract

Autoradiography has been used in the past to locate presumptive receptor systems for a number of steroids. It has provided information in rather complex tissues that could not have been obtained by biochemical procedures. In these studies we made use of autoradiography to redirect our biochemical efforts to study androgen receptors in the primate heart. Castrated-adrenalectomized female rhesus monkeys and baboons were injected with 1 microgram and 5 alpha-dihydro-[1,2,4,5,6,7-3H]-testosterone per kilogram of body weight. The animals were killed 1 hour later and the hearts were removed and processed for autoradiography. A nuclear localization of androgen was found in most of the myocardial cells, but in few if any of the interstitial cells. After reviewing the autoradiographic data, we began a new series of biochemical studies using a new buffer system and unlabeled androgen to stabilize the receptor. This in combination with postlabeled gradients allowed us to demonstrate for the first time 8S binding on sucrose density gradients. We feel that autoradiography can be a useful adjunct to biochemical studies even in "less complex" tissue such as the cardiac muscle.

摘要

过去,放射自显影术已被用于定位多种类固醇的假定受体系统。它在相当复杂的组织中提供了通过生化程序无法获得的信息。在这些研究中,我们利用放射自显影术重新引导我们的生化研究方向,以研究灵长类动物心脏中的雄激素受体。给去势-肾上腺切除的雌性恒河猴和狒狒每千克体重注射1微克5α-二氢-[1,2,4,5,6,7-³H]-睾酮。1小时后处死动物,取出心脏并进行放射自显影处理。在大多数心肌细胞中发现雄激素定位于细胞核,但在间质细胞中几乎没有发现(如果有的话)。在审查了放射自显影数据后,我们开始了一系列新的生化研究,使用新的缓冲系统和未标记的雄激素来稳定受体。这与后标记梯度相结合,使我们首次在蔗糖密度梯度上证明了8S结合。我们认为,即使在像心肌这样“不太复杂”的组织中,放射自显影术也可以成为生化研究的有用辅助手段。

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