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通过竞争性逆转录聚合酶链反应测定再生小鼠睾丸中c-myc mRNA的表达过程。

Course of c-myc mRNA expression in the regenerating mouse testis determined by competitive reverse transcriptase polymerase chain reaction.

作者信息

Amendola R

机构信息

Department of Environmental Sciences, Enea CR-Casaccia, Roma, Italy.

出版信息

DNA Cell Biol. 1994 Nov;13(11):1099-107. doi: 10.1089/dna.1994.13.1099.

Abstract

The c-myc proto-oncogene is a reliable marker of the "G0-early G1" transition, and its down-regulation is believed to be necessary to obtain cellular differentiation. In murine spermatogenesis, the level of c-myc transcripts does not correlate with the rate of cellular division. Proliferation of supposed staminal spermatogonia to reproduce themselves is induced with a local 5 Gy X-ray dose in 90-day-old C57Bl/6 mice. c-myc quantification by a newly developed competitive reverse transcriptase polymerase chain reaction (RT-PCR) was carried out to follow the expression course of this proto-oncogene. Damage and restoration of spermatogenesis were analyzed at days 3, 6, 9, 10, 13, 30, and 60 after injury by relative testes/body weight determination and histological examination. Proliferative status was determined by histone H3 Northern blot analysis. c-myc mRNA level was 10 times higher after 3 days in the irradiated animals compared to the controls. An increasing number of copies were noted up to 10 days, but promptly decreased to the base level found for irradiated mice from 13 to 60 days. Interestingly, the expression of histone H3 detected S phase only in testes at 60 days from damage.

摘要

c-myc原癌基因是“G0期至G1早期”转变的可靠标志物,其下调被认为是细胞分化所必需的。在小鼠精子发生过程中,c-myc转录本水平与细胞分裂速率无关。用局部5 Gy X射线剂量诱导90日龄C57Bl/6小鼠中假定的雄性精原细胞增殖以进行自我复制。通过新开发的竞争性逆转录聚合酶链反应(RT-PCR)对c-myc进行定量,以追踪该原癌基因的表达过程。在损伤后第3、6、9、10、13、30和60天,通过相对睾丸/体重测定和组织学检查分析精子发生的损伤和恢复情况。通过组蛋白H3 Northern印迹分析确定增殖状态。与对照组相比,照射动物在3天后c-myc mRNA水平高10倍。在第10天之前观察到拷贝数增加,但从第13天到60天迅速降至照射小鼠的基础水平。有趣的是,组蛋白H3的表达仅在损伤后60天的睾丸中检测到S期。

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