Krawczyk Z, Mali P, Parvinen M
Department of Anatomy, University of Turku, Finland.
J Cell Biol. 1988 Oct;107(4):1317-23. doi: 10.1083/jcb.107.4.1317.
Changes in the level of a testis-specific hsp70 gene-related transcript (hst70 RNA) and its cellular localization during the cycle of rat seminiferous epithelium have been investigated. Segments of seminiferous tubules at defined stages of the cycle were isolated in living condition by transillumination-assisted microdissection and the exact stages identified by phase-contrast microscopy of live cell squashes. The levels of the hst70 RNA were determined by Northern and slot blotting of whole cell lysates. High levels were found in stages XII-XIV and I to early VII of the cycle, and low levels were found in other stages, i.e., late VII (VIId) through VIII-XI of the cycle. The in situ hybridization revealed that the hst70 gene was activated in late pachytene primary spermatocytes during stage XII of the cycle, and that mRNA was then present in cells during differentiation through diakinesis, meiotic divisions, and early spermiogenesis (steps 1 through early 7). The activation of the gene coding for hst70 RNA shortly before meiotic divisions may indicate that the gene product is needed either during differentiation of late spermatocytes into spermatids or later during spermiogenesis, and that the mRNA may be stored in early spermatids.
研究了大鼠生精上皮周期中睾丸特异性热休克蛋白70基因相关转录本(hst70 RNA)水平的变化及其细胞定位。通过透照辅助显微切割在活体条件下分离出处于周期特定阶段的生精小管片段,并通过活细胞压片的相差显微镜确定确切阶段。通过全细胞裂解物的Northern印迹和狭缝印迹法测定hst70 RNA的水平。在周期的XII - XIV期以及I期至VII期早期发现高水平,而在其他阶段,即周期的VII期晚期(VIId)至VIII - XI期发现低水平。原位杂交显示,hst70基因在周期的XII期晚期粗线期初级精母细胞中被激活,然后在细胞从终变期、减数分裂到精子发生早期(第1步至早期第7步)的分化过程中mRNA都存在。减数分裂前不久编码hst70 RNA的基因被激活,这可能表明该基因产物在晚期精母细胞分化为精子细胞的过程中或在精子发生后期是必需的,并且mRNA可能储存在早期精子细胞中。