Baker P J, O'Shaughnessy P J
Division of Veterinary Physiology and Pharmacology, Department of Veterinary Preclinical Studies, University of Glasgow Veterinary School, Bearsden Rd, Glasgow G61 1QH, UK.
Reproduction. 2001 Oct;122(4):553-9. doi: 10.1530/rep.0.1220553.
Prostaglandin D synthetase is expressed relatively highly in the testis and reproductive tract of a number of species, including the mouse. In adult mouse testis, expression is confined largely to the Leydig cells and in this study changes in the expression and localization of prostaglandin D synthetase mRNA during testis development were examined. Initial studies using RT-PCR and isolated testicular compartments indicated that prostaglandin D synthetase expression in the neonatal testis was predominantly within the seminiferous tubules. In situ hybridization studies confirmed that prostaglandin D synthetase mRNA appears to be expressed only in the tubules of neonatal mouse testes and only in the interstitial tissue of the adult testis. TaqMan real-time PCR was used to quantify prostaglandin D synthetase mRNA content during development using an exogenous mRNA as a control standard. Expression per testis decreased after birth to < 10% at day 15 before recovering again by days 25-30. After day 30, expression per testis increased 40-fold during final development to adulthood. Studies using RT-PCR showed that early expression before day 15 was restricted to the tubular compartment, whereas the subsequent increase in expression after day 30 was restricted to the interstitial compartment. Database analysis showed that the 3' end of the prostaglandin D synthetase transcript was subject to alternate splicing. Both splice isoforms were shown by RT-PCR to be present throughout development and without a major change in expression pattern. These results indicate that expression of prostaglandin D synthetase mRNA shifts during development from the tubular compartment of the fetal or neonatal testis to the developing adult Leydig cells, with expression in the Leydig cells increasing markedly after puberty. These changes are similar to those observed for 17beta-hydroxysteroid dehydrogenase type III and may indicate that this developmental process is not uncommon in the testis.
前列腺素D合成酶在包括小鼠在内的许多物种的睾丸和生殖道中表达相对较高。在成年小鼠睾丸中，表达主要局限于睾丸间质细胞，在本研究中，检测了睾丸发育过程中前列腺素D合成酶mRNA的表达和定位变化。最初使用逆转录聚合酶链反应（RT-PCR）和分离的睾丸区室进行的研究表明，新生小鼠睾丸中前列腺素D合成酶的表达主要在生精小管内。原位杂交研究证实，前列腺素D合成酶mRNA似乎仅在新生小鼠睾丸的小管中表达，且仅在成年睾丸的间质组织中表达。使用TaqMan实时PCR，以一种外源mRNA作为对照标准，对发育过程中前列腺素D合成酶mRNA的含量进行定量。出生后每个睾丸的表达下降，在第15天时降至<10%，然后在第25 - 30天再次恢复。第30天后，每个睾丸的表达在最终发育至成年期的过程中增加了40倍。使用RT-PCR的研究表明，第15天之前的早期表达局限于小管区室，而第30天之后表达的随后增加则局限于间质区室。数据库分析表明，前列腺素D合成酶转录本的3'端存在可变剪接。逆转录聚合酶链反应显示两种剪接异构体在整个发育过程中均存在，且表达模式无重大变化。这些结果表明，前列腺素D合成酶mRNA的表达在发育过程中从胎儿或新生小鼠睾丸的小管区室转移至发育中的成年睾丸间质细胞，且间质细胞中的表达在青春期后显著增加。这些变化与在III型17β-羟基类固醇脱氢酶中观察到的变化相似，可能表明这种发育过程在睾丸中并不罕见。
