Expression of protamine-1 and-2 mRNA during human spermiogenesis

被引:103
作者
Steger, K
Pauls, K
Klonisch, T
Franke, FE
Bergmann, M
机构
[1] Inst Vet Anat, D-35392 Giessen, Germany
[2] Inst Pathol, D-35392 Giessen, Germany
[3] Inst Anat & Zellbiol, D-06097 Halle, Germany
关键词
human testis; in-situ hybridization; laser-assisted cell-picking; protamines; spermiogenesis;
D O I
10.1093/molehr/6.3.219
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
During spermiogenesis, the histone-to-protamine replacement causes the compaction of the spermatid chromatin. The genes for protamines, PRM-1 and PRM-2, are transcribed in round and elongating spermatids, The transcripts are stored in a translationally-repressed state by the binding of protein repressors before being translated in elongating and elongated spermatids. RNA extracts from homogenized whole testis samples supply only average data, and cell-specific and stage-specific expression cannot be addressed. Therefore, we used UV-laser-assisted cell-picking (UV-LACP) to select spermatids of defined differentiation steps, Subsequent reverse transcription-polymerase chain reaction (RT-PCR) with intron-spanning primer pairs allowed the detection of DNA-free and pseudogene-free PRM-1 and PRM-P cDNA. Additional in-situ hybridization with digoxygenin-labelled cRNA probes exhibited PRM-1 and PRM-2 mRNA from step 1/2 spermatids to step 4 spermatids, but not in elongated spermatids. RT-PCR revealed amplicons for PRIM-1 and PRM-2 in all spermatids except step 3 round spermatids, Applying proteinase K digestion, PRM-1 and PRM-2 transcripts were also detected in step 3 spermatids indicating that protein repressors may bind to both PRM-1 and PRM-2 mRNA in step 3 round spermatids. These data demonstrate that the combination of UV-LACP and non-radioactive in-situ hybridization appear to be a suitable approach for the study of cell-specific and stage-specific gene expression during spermiogenesis.
引用
收藏
页码:219 / 225
页数:7
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