大鼠脑cDNA编码具有酶活性的γ-氨基丁酸转氨酶，并为γ-氨基丁酸分解代谢细胞提供了一种分子探针。

A rat brain cDNA encodes enzymatically active GABA transaminase and provides a molecular probe for GABA-catabolizing cells.

作者信息

Medina-Kauwe L K, Tillakaratne N J, Wu J Y, Tobin A J

机构信息

Molecular Biology Institute, University of California-Los Angeles 90024-1606.

出版信息

J Neurochem. 1994 Apr;62(4):1267-75. doi: 10.1046/j.1471-4159.1994.62041267.x.

DOI:10.1046/j.1471-4159.1994.62041267.x

PMID:8133261

Abstract

cDNAs encoding gamma-aminobutyric acid aminotransferase (GABA-T) were isolated from a lambda ZAP rat hippocampal cDNA expression library by two independent cloning methods, immunological screening with an antimouse GABA-T antibody and plaque hybridization with a GABA-T cDNA probe derived by polymerase chain reaction. We have produced enzymatically active GABA-T from a rat brain cDNA containing the full-length GABA-T coding region. Our rat brain GABA-T cDNAs hybridize to mRNAs in brain and peripheral tissues, including liver, kidney, and testis. We have also detected GABA-T mRNA in GABAergic cells of rat cerebellar cortex by in situ hybridization. Our rat brain GABA-T probe hybridizes to Purkinje, basket, stellate, and Golgi II cells, the same GABAergic neurons previously shown to contain glutamate decarboxylase GAD65 and GAD67.

摘要

通过两种独立的克隆方法，即使用抗小鼠γ-氨基丁酸转氨酶（GABA-T）抗体进行免疫筛选以及用聚合酶链反应衍生的GABA-T cDNA探针进行噬菌斑杂交，从λZAP大鼠海马cDNA表达文库中分离出编码GABA-T的cDNA。我们已经从包含全长GABA-T编码区的大鼠脑cDNA中产生了具有酶活性的GABA-T。我们的大鼠脑GABA-T cDNA与脑和外周组织（包括肝脏、肾脏和睾丸）中的mRNA杂交。我们还通过原位杂交在大鼠小脑皮质的GABA能细胞中检测到了GABA-T mRNA。我们的大鼠脑GABA-T探针与浦肯野细胞、篮状细胞、星状细胞和高尔基II型细胞杂交，这些都是先前显示含有谷氨酸脱羧酶GAD65和GAD67的相同GABA能神经元。