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大鼠肾脏中肾上腺素能受体的放射自显影定量分析。

Autoradiographic quantification of adrenergic receptors in rat kidney.

作者信息

Calianos T, Muntz K H

机构信息

Department of Cell Biology and Neurosciences, University of Texas Southwestern Medical Center, Dallas.

出版信息

Kidney Int. 1990 Jul;38(1):39-46. doi: 10.1038/ki.1990.164.

Abstract

The adrenergic nervous system is active in kidney function, and the kidney has large numbers of adrenergic receptor subtypes. Because of the cellular complexity of the kidney, it is difficult to obtain direct assessments of adrenergic receptor binding characteristics over specific tissue compartments. Qualitative autoradiography allows the localization of adrenergic receptors over tissue types in the kidney, but quantitative autoradiography allows direct comparison of adrenergic receptor number over different cellular compartments. The purpose of this study was to obtain direct assessments of alpha 1, alpha 2, and beta adrenergic receptor numbers over different tissue compartments of the kidney using quantitative autoradiography. Sections of Sprague-Dawley rat kidney were incubated in several concentrations of 3H-dihydroalprenolol to label beta receptors, 3H-prazosin to label alpha 1 receptors and 3H-rauwolscine to label the alpha 2 receptors. Sections of rat heart incubated in 3H-dihydroalprenolol were included as standards. The sections were then prepared for receptor autoradiography. After processing, the grains were then quantified on an image analysis system, and binding curves constructed from the specific binding. In some animals, the proximal tubules were stained to localize the proximal convoluted tubules. Significant Scatchard analyses were obtained in the glomeruli with dihydroalprenolol (5.18 X 10(9) receptors/mm3) and with rauwolscine (2.48 X 10(9) receptors/mm3). Significant Scatchard analyses were obtained in the cortex with rauwolscine (9.47 X 10(9) receptors/mm3) and with prazosin (3.9 X 10(9)). In addition, specific binding was seen with rauwolscine and prazosin to the kidney arterioles.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

肾上腺素能神经系统在肾功能中发挥作用,且肾脏有大量肾上腺素能受体亚型。由于肾脏细胞的复杂性,难以直接评估特定组织区域内肾上腺素能受体的结合特性。定性放射自显影可确定肾脏中不同组织类型上肾上腺素能受体的定位,但定量放射自显影能直接比较不同细胞区域内肾上腺素能受体的数量。本研究的目的是使用定量放射自显影直接评估肾脏不同组织区域内α1、α2和β肾上腺素能受体的数量。将斯普拉格-道利大鼠肾脏切片在几种浓度的3H-二氢阿普洛尔中孵育以标记β受体,在3H-哌唑嗪中孵育以标记α1受体,在3H-萝芙木碱中孵育以标记α2受体。将在3H-二氢阿普洛尔中孵育的大鼠心脏切片作为标准物。然后对切片进行受体放射自显影准备。处理后,在图像分析系统上对颗粒进行定量,并根据特异性结合构建结合曲线。在一些动物中,对近端小管进行染色以定位近端曲管。在肾小球中,用二氢阿普洛尔(5.18×10⁹受体/mm³)和萝芙木碱(2.48×10⁹受体/mm³)获得了显著的斯卡查德分析结果。在皮质中,用萝芙木碱(9.47×10⁹受体/mm³)和哌唑嗪(3.9×10⁹)获得了显著的斯卡查德分析结果。此外,观察到萝芙木碱和哌唑嗪与肾小动脉有特异性结合。(摘要截短于250字)

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