Developmental expression of the S35-S45/SGP-2/TRPM-2 gene in rat testis and epididymis.
Developmental expression of the S35-S45/SGP-2/TRPM-2 gene in rat testis and epididymis.
复制标题
S35-S45/SGP-2/TRPM-2 基因在大鼠睾丸和附睾中的发育表达。
DOI：
10.1002/mrd.1080330403
复制
发表时间：
1992
影响因子：
2.5
通讯作者：
Tenniswood,M
中科院分区：
文献类型：
--
作者：
Zakeri,Z;Curto,M;Hoover,D;Wightman,K;Engelhardt,J;Smith,FF;Kierszenbaum,AL;Gleeson,T;Tenniswood,M
睾酮抑制的前列腺信息-2(TRPM-2)最初是从大鼠退行性前列腺腹侧分离和克隆的。在这个组织中，以及在其他依赖激素的组织中，如乳腺，这种基因在缺乏适当的营养激素的情况下被诱导。序列分析表明，TRPM-2基因的编码序列与S35-S45(又称SGP-2和Clusterin)完全相同，S35-S45是由成人睾丸支持细胞结构性表达的。利用Northern杂交、狭缝杂交、S1-核酸酶分析和原位杂交等方法，研究了TRPM-2在睾丸和附睾发育过程中的表达规律。从7日龄、14日龄、28日龄、35日龄和91日龄大鼠睾丸和附睾提取的RNA的狭缝印迹分析表明，该基因在7日龄到14日龄之间被诱导到可检测到的水平，而相对表达水平在14日龄后没有显著变化。用2、7、14、28、35和91日龄大鼠睾丸冰冻切片的原位杂交证实，TPRM-2在7日龄大鼠生精上皮中几乎没有表达，但在14日龄后表达显著增加。主要在支持细胞中，但也与减数分裂发育中的生精细胞有关。然而，TRPM-2mRNA在2日龄时在睾丸网部表达，35日龄时达到高峰，并在成年后继续表达。狭缝印迹分析表明，TRPM-2在7日龄至14日龄的附精中也被诱导表达，但正如原位杂交所证明的那样，TRPM-2mRNA在附睾头的上皮细胞中可检测到，而在附睾部或尾部几乎检测不到。Northern分析表明，在发育过程中，睾丸中TRPM-2转录本的大小也会发生变化。在睾丸发育的早期阶段，TRPM-2的转录本似乎是一条约1.5kb的宽带，而成年的转录本似乎有大约1.8kb的长度。S1-核酸酶保护分析表明，这种大小的增加并不是由于TRPM-2/SGP-2第一个外显子的差异剪接，最可能反映的是发育过程中不同时期睾丸mRNA多聚腺苷化的差异。©1992 Wiley-Liss，Inc.
Testosterone‐repressed prostate message‐2 (TRPM‐2) was originally isolated and cloned from the regressing ventral prostate of the rat. In this tissue, and in other hormone‐dependent tissues such as the mammary gland, this gene is induced in the absence of the appropriate trophic hormone. Sequence analysis of the cDNA and genomic clones of TRPM‐2 have demonstrated that the coding sequence of this gene is identical to S35‐S45 (also known as SGP‐2 and clusterin), which is constitutively expressed by the Sertoli cells of the adult testis. Using Northern, slot blot, S1‐nuclease analysis, and in situ hybridization, we have investigated the regulation of TRPM‐2 expression in the testis and epididymis during development. Slot blot analysis of RNA extracted from the testis and epididymis of 7‐, 14‐, 28‐, 35‐, and 91‐day‐old rats demonstrates that the gene is induced to detectable levels between days 7 and 14 and that the relative level of expression does not change significantly after day 14. In situ hybridization using frozen sections of testis from day 2‐, 7‐, 14‐, 28‐, 35‐, and 91‐day‐old rats confirms that there is little expression of TPRM‐2 in the seminiferous epithelium of 7‐day‐old rats, but this increases considerably after 14 days, primarily in Sertoli cells but also in association with meiotic developing spermatogenic cells. However, TRPM‐2 mRNA is expressed in the rete testis at 2 days of age, reaches a peak at 35 days of age, and continues to be expressed in the adult. Slot blot analysis demonstrates that TRPM‐2 is also induced in the epididymis between 7 and 14 days of age, although, as has been demonstrated by in situ hybridization, TRPM‐2 mRNA is detectable in the epithelial cells in the head of the epididymis but is barely detectable in the midportion or tail regions. Northern analysis suggests that the size of the TRPM‐2 transcript in the testis also changes during development. In the early stages of testicular development, the TRPM‐2 transcript appears to be a broad band of approximately 1.5 kb, while the transcript in the adult appears to be approximately 1.8 kb in length. S1‐nuclease protection assays suggest that this increase in size is not due to differential splicing of the first exon of TRPM‐2/SGP‐2 and most probably reflects a difference in the polyadenylation of the mRNA in the testis at different times during development. © 1992 Wiley‐Liss, Inc.