Marret C, Avallet O, Perrard-Sapori M H, Durand P
INSERM-INRA, Hôpital Debrousse, Lyon, France.
Mol Reprod Dev. 1998 Sep;51(1):22-35. doi: 10.1002/(SICI)1098-2795(199809)51:1<22::AID-MRD3>3.0.CO;2-Y.
Expression of the testis-specific histone TH2B, the phosphoprotein p19, and the transition proteins TP1 and TP2, was localized in the rat testis and quantified, using in situ hybridization of their mRNAs with radiolabeled probes and image analysis. In a first study, expression was assessed during testicular development between day 2 and day 65 postpartum. TH2B mRNAs appeared first in preleptotene spermatocytes (PL) on day 12 and in pachytene spermatocytes (PS) on day 18; p19 mRNAs were present in PS from day 18 onward, and TP1 and TP2 mRNAs were detected in round spermatids (RS) from day 32 onward. In the second trial, the expression of these four genes was studied throughout the cycle of spermatogenic epithelium in mature animals. TH2B mRNAs were localized in B spermatogonia at stage V, and in PL at stages VII and VIII but no longer in leptotene and zygotene spermatocytes. Thereafter, TH2B mRNAs were observed in PS from stages III-IV to XIII. The steady-state mRNA level per cell was high in PS with a maximum at stages IX-X. p19 mRNAs were present in PS from stages III-IV onward and in RS up to stages 1-2 of spermiogenesis. The maximum mRNA level per cell was observed in PS between stages VII and XIII. The presence of TP1 mRNAs was restricted to spermatids from steps 6 to 15-16 of spermiogenesis while TP2 mRNAs were detected in spermatids only between step 7 and step 13. The highest steady-state amounts of mRNAs were observed between step 7 and step 14 for TP1 and between step 10 and step 12 for TP2.
利用放射性标记探针原位杂交技术及其图像分析,对睾丸特异性组蛋白TH2B、磷蛋白p19以及过渡蛋白TP1和TP2在大鼠睾丸中的表达进行了定位和定量分析。在第一项研究中,评估了产后第2天至第65天睾丸发育过程中的表达情况。TH2B mRNA在产后第12天首次出现在前细线期精母细胞(PL)中,第18天出现在粗线期精母细胞(PS)中;p19 mRNA从第18天起存在于PS中,TP1和TP2 mRNA从第32天起在圆形精子细胞(RS)中被检测到。在第二项试验中,研究了成熟动物生精上皮周期中这四个基因的表达情况。TH2B mRNA定位于Ⅴ期的B型精原细胞以及Ⅶ期和Ⅷ期的PL,但不再定位于细线期和偶线期精母细胞。此后,在Ⅲ - Ⅳ期至ⅩⅢ期的PS中观察到TH2B mRNA。每个细胞的稳态mRNA水平在PS中较高,在Ⅸ - Ⅹ期达到最大值。p19 mRNA从Ⅲ - Ⅳ期起存在于PS中,在精子发生的1 - 2期的RS中也有存在。每个细胞的最大mRNA水平在Ⅶ期至ⅩⅢ期的PS中观察到。TP1 mRNA仅存在于精子发生第6步至15 - 16步的精子细胞中,而TP2 mRNA仅在第7步至第13步的精子细胞中被检测到。TP1的最高稳态mRNA量在第7步至第14步之间观察到,TP2的最高稳态mRNA量在第10步至第12步之间观察到。