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Mouse testes contain two size classes of actin mRNA that are differentially expressed during spermatogenesis.

机译:小鼠睾丸包含两个大小类别的肌动蛋白mRNA在精子发生过程中差异表达。

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摘要

Using several actin isotype-specific cDNA probes, we found actin mRNA of two size classes, 2.1 and 1.5 kilobases (kb), in extracts of polyadenylated and nonpolyadenylated RNA from sexually mature CD-1 mouse testes. Although the 2.1-kb sequence was present in both meiotic and postmeiotic testicular cell types, it decreased manyfold in late haploid cells. The 1.5-kb actin sequence was not detectable in meiotic pachytene spermatocytes (or in liver or kidney cells), but was present in round and elongating spermatids and residual bodies. To differentiate between the beta- and gamma-actin mRNAs, we isolated a cDNA, pMGA, containing the 3' untranslated region of a mouse cytoplasmic actin that has homology to the 3' untranslated region of a human gamma-actin cDNA but not to the 3' untranslated regions of human alpha-, beta-, or cardiac actins. Dot blot hybridizations with pMGA detected high levels of presumptive gamma-actin mRNA in pachytene spermatocytes and round spermatids, with lower amounts found in elongating spermatids. Hybridization with the 3' untranslated region of a rat beta-actin probe revealed that round spermatids contained higher levels of beta-actin mRNA than did pachytene spermatocytes or residual bodies. Both probes hybridized to the 2.1-kb actin mRNA but failed to hybridize to the 1.5-kb mRNA.
机译:使用几种肌动蛋白同种型特异性cDNA探针,我们从性成熟的CD-1小鼠睾丸的聚腺苷酸化和非聚腺苷酸化RNA的提取物中发现了两个大小级别的肌动蛋白mRNA,分别为2.1和1.5千碱基。尽管在减数分裂和后减数分裂的睾丸细胞类型中均存在2.1-kb序列,但在晚期单倍体细胞中它减少了许多倍。 1.5 kb肌动蛋白序列在减数分裂的粗线精子细胞(或肝或肾细胞)中未检出,但存在于圆形的和延伸的精子细胞和残体中。为了区分β-和γ-肌动蛋白的mRNA,我们分离了一个cDNA pMGA,其中包含小鼠细胞质肌动蛋白的3'非翻译区域,该区域与人γ-actincDNA的3'非翻译区域具有同源性,但与人类α-,β-或心脏肌动蛋白的3'非翻译区。与pMGA的斑点印迹杂交在粗线精子细胞和圆形精子细胞中检测到高水平的推定的γ-肌动蛋白mRNA,而在伸长的精子细胞中发现的含量较低。与大鼠β-肌动蛋白探针的3'非翻译区杂交后发现,圆形精子中的β-肌动蛋白mRNA水平高于粗线精原细胞或残体。两种探针均与2.1-kb肌动蛋白mRNA杂交,但未能与1.5-kb mRNA杂交。

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