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通过定量PCR检测小鼠精子发生过程中次黄嘌呤磷酸核糖基转移酶(HPRT)、磷酸甘油酸激酶-1(PGK-1)、磷酸甘油酸激酶-2(PGK-2)、腺嘌呤磷酸核糖转移酶(APRT)、甲基转移酶(MTase)和Zfy基因转录本的变化。

Measurement by quantitative PCR of changes in HPRT, PGK-1, PGK-2, APRT, MTase, and Zfy gene transcripts during mouse spermatogenesis.

作者信息

Singer-Sam J, Robinson M O, Bellvé A R, Simon M I, Riggs A D

机构信息

Division of Biology, Beckman Research Institute, City of Hope, Duarte, CA 91010.

出版信息

Nucleic Acids Res. 1990 Mar 11;18(5):1255-9. doi: 10.1093/nar/18.5.1255.

Abstract

A reverse transcriptase-polymerase chain reaction assay (RT-PCR) was used quantitatively to measure accumulated levels of RNA transcripts in total mouse RNAs derived from male germ cells at various spermatogenic stages. RNA levels for two X-linked enzymes, phosphoglycerate kinase (PGK-1) and hypoxanthine phosphoribosyl transferase (HPRT), both decrease during spermatogenesis, although the transcript levels decrease much more rapidly for PGK-1. RNA for the Y-linked ZFY (zinc finger protein) is elevated in all spermatogenic cell fractions tested, being particularly high in leptotene/zygotene spermatocytes and round spermatids. RNA for adenine phosphoribosyltransferase (APRT) increases 5-fold to a peak during late pachynema. RNA for PGK-2, undetectable in spermatogonial cells, increases at least 50-fold by the round spermatid stage. DNA (cytosine-5-)-methyltransferase (MTase) transcript levels are over an order of magnitude higher throughout spermatogenesis than in non-dividing liver cells.

摘要

采用逆转录聚合酶链反应分析(RT-PCR)定量测定了来自不同生精阶段雄性生殖细胞的小鼠总RNA中RNA转录本的积累水平。两种X连锁酶,磷酸甘油酸激酶(PGK-1)和次黄嘌呤磷酸核糖转移酶(HPRT)的RNA水平在精子发生过程中均下降,尽管PGK-1的转录本水平下降得更快。Y连锁的ZFY(锌指蛋白)的RNA在所有测试的生精细胞组分中均升高,在细线期/偶线期精母细胞和圆形精子细胞中尤其高。腺嘌呤磷酸核糖转移酶(APRT)的RNA在粗线期后期增加5倍达到峰值。PGK-2的RNA在精原细胞中无法检测到,到圆形精子细胞阶段至少增加50倍。DNA(胞嘧啶-5-)-甲基转移酶(MTase)的转录本水平在整个精子发生过程中比不分裂的肝细胞高一个数量级以上。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/500a/330442/438f527dd813/nar00189-0156-a.jpg

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