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Acta Biochimica et Biophysica Sinica Advance Access originally published online on May 29, 2009
Acta Biochimica et Biophysica Sinica 2009 41(7):545-553; doi:10.1093/abbs/gmp043
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© The Author 2009. Published by ABBS Editorial Office in association with Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.

Dynamic regulation of glutamic acid decarboxylase 65 gene expression in rat testis

Haixiong Liu{dagger}, Shifeng Li{dagger}, Yunbin Zhang, Yuanchang Yan and Yiping Li*

Shanghai Key Laboratory for Molecular Andrology, Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Graduate School of the Chinese Academy of Sciences, Shanghai 200031, China

* Correspondence address. Tel: +86-21-5492139; Fax: +86-21-54921415; E-mail: yipingli{at}sibs.ac.cn


  Abstract

Glutamate decarboxylase 65 (GAD65) produces {gamma}-aminobutyric acid, the main inhibitory neurotransmitter in adult mammalian brain. Previous experiments, performed in brain, showed that GAD65 gene possesses two TATA-less promoters, although the significance is unknown. Here, by rapid amplification of cDNA ends method, two distinct GAD65 mRNA isoforms transcribed from two independent clusters of transcription start sites were identified in post-natal rat testis. RT–PCR results revealed that the two mRNA isoforms had distinct expression patterns during post-natal testis maturation, suggesting that GAD65 gene expression was regulated by alternative promoters at the transcription level. By using GAD65-specific antibodies, western blotting analysis showed that the 58-kDa GAD65, N-terminal 69 amino acids truncated form of full-length GAD65 protein, was developmentally expressed during post-natal testis maturation, suggesting that GAD65 gene expression in testis may also be regulated by post-translational processing. Confocal immunofluorescence microscopy revealed that GAD65 protein was presented in Leydig cells of Day 1 testis, primary spermatocytes and spermatids of post-natal of Day 90 testis. The above results suggested that GAD65 gene expression is dynamically regulated at multiple levels during post-natal testis maturation.

Keywords    transcription; gene regulation; glutamate decarboxylase; testis

Received: February 16, 2009; Accepted: May 5, 2009

{dagger} These authors contributed equally to this work.


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