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使用35S标记的RNA探针通过原位杂交对大鼠肾脏中γ-谷氨酰转肽酶mRNA进行定位。

Localization by in situ hybridization of gamma-glutamyl transpeptidase mRNA in the rat kidney using 35S-labeled RNA probes.

作者信息

Lebargy F, Bulle F, Siegrist S, Guellaen G, Bernaudin J F

机构信息

Unités INSERM 139 et 99, Département d'Histologie, Faculté de Médecine, Université Paris, Creteil, France.

出版信息

Lab Invest. 1990 Jun;62(6):731-5.

PMID:1972767
Abstract

gamma-Glutamyl transpeptidase (GGT) is involved in the extraction of plasma glutathione in the postglomerular compartment of the kidney. The enzyme is distributed in the proximal tubule associated with the apical brush border. Using in situ hybridization with 35S-labeled anti-sense and sense RNA GGT probes on rat kidney cryostat sections, the present study demonstrates that GGT mRNA transcripts are detected in proximal tubules localized in the inner cortex, outer medulla, and medullary rays of the cortex. Such a distribution suggests that the GGT gene is mainly expressed in the more distal part of the proximal tubule, i.e., the pars recta. As a control, in situ hybridization has also been performed using a 35S-labeled beta-actin anti-sense RNA probe showing a diffuse pattern of distribution of beta-actin RNA transcripts particularly in the outer cortex. This highly sensitive method using single strand asymetric RNA probes, with a markedly reduced nonspecific binding is promising for the study of gene expression patterns in the kidney in order to clarify their heterogeneity in the various segments of the nephron.

摘要

γ-谷氨酰转肽酶(GGT)参与肾脏肾小球后段血浆谷胱甘肽的提取过程。该酶分布于近端小管,与顶端刷状缘相关。本研究采用35S标记的反义及正义RNA GGT探针,对大鼠肾脏冰冻切片进行原位杂交,结果显示在位于肾皮质内带、外髓质及皮质髓放线的近端小管中可检测到GGT mRNA转录本。这种分布表明GGT基因主要在近端小管的更远端部分,即直部表达。作为对照,还使用35S标记的β-肌动蛋白反义RNA探针进行了原位杂交,结果显示β-肌动蛋白RNA转录本呈弥漫性分布模式,尤其在外皮质更为明显。这种使用单链不对称RNA探针的高灵敏度方法,非特异性结合明显减少,有望用于研究肾脏中的基因表达模式,以阐明其在肾单位各节段中的异质性。

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