Quantitation of mRNAs during mouse spermatogenesis: protamine-like histone and phosphoglycerate kinase-2 mRNAs increase after meiosis.
Quantitation of mRNAs during mouse spermatogenesis: protamine-like histone and phosphoglycerate kinase-2 mRNAs increase after meiosis.
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小鼠精子发生过程中 mRNA 的定量：鱼精蛋白样组蛋白和磷酸甘油酸激酶 2 mRNA 在减数分裂后增加。
DOI：
10.1073/pnas.77.10.6086
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发表时间：
1980
影响因子：
11.1
通讯作者：
A. Salkeld
中科院分区：
文献类型：
--
作者：
R. Erickson;J. Kramer;J. Rittenhouse;A. Salkeld
从未成熟或成熟小鼠睾丸或从根据沉降速度分离的生精细胞制备总RNA，进行体外翻译。以[~ 3 H]精氨酸为标记物，小鼠鱼精蛋白样组蛋白可被鉴定为体外翻译产物。鱼精蛋白样组蛋白的mRNA仅在减数分裂后检测到;只有当成熟睾丸或晚期精子细胞组分的RNA被翻译时，才出现与鱼精蛋白样组蛋白共迁移的放射性峰。当[35 S]甲硫氨酸为标记物时，通过亲和色谱法随后进行二维凝胶电泳或通过特异性免疫沉淀法将磷酸甘油酸激酶-2（ATP：3-磷酸-D-甘油酸I-磷酸转移酶，EC 2.7.2.3）鉴定为体外翻译产物。磷酸甘油酸激酶-2的mRNA仅在成熟睾丸或晚期精子细胞组分中检测到。这些鱼精蛋白样组蛋白和磷酸甘油酸激酶-2 mRNA的翻译试验表明，这些信息在减数分裂后转录。
Total RNA, prepared from immature or mature mouse testes or from spermatogenic cells separated on the basis of sedimentation velocity, was translated in vitro. Mouse protamine-like histone could be identified as an in vitro translational product when [3H]arginine was used as the label. The mRNA for protamine-like histone was detected only after meiosis; the appearance of a peak of radioactivity comigrating with protamine-like histone occurred only when RNA from mature testes or late spermatid cell fractions was translated. Phosphoglycerate kinase-2 (ATP:3-phospho-D-glycerate I-phosphotransferase, EC 2.7.2.3) was identified as an in vitro translational product by affinity chromatography followed by two-dimensional gel electrophoresis or by specific immunoprecipitation when [35S]methionine was the label. The mRNA for phosphoglycerate kinase-2 was detected only in mature testes or late spermatid cell fractions. These translational assays for protamine-like histone and phosphoglycerate kinase-2 mRNAs suggest that these messages are transcribed after meiosis.