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大鼠附睾起始段γ-谷氨酰转肽酶mRNA表达的稳定性及转录调控

Stability and transcriptional regulation of gamma-glutamyl transpeptidase mRNA expression in the initial segment of the rat epididymis.

作者信息

Rudolph D B, Hinton B T

机构信息

Department of Cell Biology and The Center for Research in Reproduction, University of Virginia, Health Sciences Center, Charlottesville 22908, USA.

出版信息

J Androl. 1997 Sep-Oct;18(5):501-12.

PMID:9349748
Abstract

Gamma-glutamyl transpeptidase (GGT) is an enzyme believed to play a role in the protection of maturing spermatozoa in the epididymis. Our previous studies have shown that four GGT mRNAs (I-IV) transcribed from the single-copy rat GGT gene are differentially expressed and regulated in the rat epididymis. In particular, the normal expression of GGT mRNA(IV) in the epididymal initial segment is dependent upon the presence of testicular factors. The objective of this study was to test the hypothesis that the decreased expression of GGT mRNA(IV) in the initial segment following the in vivo removal of testicular factors by efferent duct ligation (EDL) is due to a decrease in stability and/or transcription rate. The stability of the GGT mRNAs was evaluated by measuring the rate of mRNA decay. These stability studies showed that GGT mRNA(IV) exhibited a rapid initial decay that slowed at later times to a decay rate similar to that of GGT mRNAs(II,III). The decay rates were not different following sham-operation or EDL, and thus the stability of GGT mRNAs were not influenced by the in vivo loss of testicular factors. Results of transcription analysis revealed that the transcription rate of GGT mRNA(IV) in the initial segment fell by approximately 68% following a 12-hour EDL. Additionally, secondary-structure models indicate two families of folding patterns for GGT mRNA(IV), which could be the reason for the two decay regimes detected in the stability study. Thus, the decreased expression level of GGT mRNA(IV) in the initial segment following the in vivo loss of testicular factors is a function of a decreased transcription rate and intricate decay kinetics.

摘要

γ-谷氨酰转肽酶(GGT)是一种据信在附睾中对成熟精子起保护作用的酶。我们之前的研究表明,从单拷贝大鼠GGT基因转录而来的四种GGT mRNA(I-IV)在大鼠附睾中差异表达且受到调控。特别是,附睾起始段中GGT mRNA(IV)的正常表达依赖于睾丸因子的存在。本研究的目的是检验以下假设:通过输出小管结扎(EDL)在体内去除睾丸因子后,附睾起始段中GGT mRNA(IV)表达的降低是由于稳定性和/或转录速率的下降。通过测量mRNA衰减率来评估GGT mRNA的稳定性。这些稳定性研究表明,GGT mRNA(IV)最初表现出快速衰减,随后衰减速度减慢,达到与GGT mRNA(II,III)相似的衰减率。假手术或EDL后的衰减率没有差异,因此GGT mRNA的稳定性不受体内睾丸因子缺失的影响。转录分析结果显示,12小时的EDL后,附睾起始段中GGT mRNA(IV)的转录速率下降了约68%。此外,二级结构模型表明GGT mRNA(IV)存在两种折叠模式家族,这可能是稳定性研究中检测到两种衰减模式的原因。因此,体内睾丸因子缺失后,附睾起始段中GGT mRNA(IV)表达水平的降低是转录速率下降和复杂衰减动力学的作用结果。

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