Jabbour H N, Clarke L A, McNeilly A S, Edery M, Kelly P A
MRC Reproductive Biology Unit, Edinburgh, UK.
J Mol Endocrinol. 1998 Apr;20(2):175-82. doi: 10.1677/jme.0.0200175.
This study investigated the pattern and site of expression of the prolactin receptor gene in the testis and epididymis of red deer collected during the breeding season (n=3). Ribonuclease protection assays using 50 microg total RNA and a 300 bp [32P]-labelled antisense cRNA probe, generated from the extracellular domain of the red deer prolactin receptor, confirmed the expression of the receptor in both the testis and epididymis; a higher level of prolactin receptor mRNA was detected in the epididymis compared with the testis (170.4+/-1.5 x 10(3) and 26.3+/-2.7 x 10(3) arbitrary units respectively; P<0.05). In situ hybridisation using 300 bp [33P]-labelled sense and antisense cRNA probes generated from the extracellular domain of the receptor localised the expression sites to the seminiferous tubules and interstitial compartments of the testis and the epithelial layer of the epididymal duct. Quantification of grain numbers demonstrated a higher level of expression of the receptor in the epididymis compared with the interstitial and seminiferous tubule compartments of the testis (18.1+/-4.4 x 10(2), 10.1+/-2.0 x 10(2) and 8.3+/-0.8 x 10(2) grains/microm2 respectively; P<0.05). However, no differences were detected in the level of expression of the receptor between the interstitial and seminiferous tubule compartments of the testis. Immunocytochemistry using an anti-prolactin receptor antibody, raised against a peptide sequence from the extracellular domain of the rat prolactin receptor, localised expression of the receptor gene to the Leydig cells, pachytene spermatocytes, round spermatids and elongating spermatids. In the epididymis, the receptor was localised to the epithelial layer within the epididymal ducts. Expression of the prolactin receptor gene in the red deer testis and epididymis suggests a role for the hormone in steroidogenesis and spermatogenesis.
本研究调查了繁殖季节采集的马鹿（n = 3）睾丸和附睾中催乳素受体基因的表达模式及表达位点。使用50微克总RNA和由马鹿催乳素受体胞外域产生的300碱基对[32P]标记反义cRNA探针进行核糖核酸酶保护分析，证实该受体在睾丸和附睾中均有表达；与睾丸相比，附睾中检测到更高水平的催乳素受体mRNA（分别为170.4±1.5×10³和26.3±2.7×10³任意单位；P<0.05）。使用由受体胞外域产生的300碱基对[33P]标记正义和反义cRNA探针进行原位杂交，将表达位点定位到睾丸的生精小管和间质区以及附睾管的上皮层。颗粒计数定量显示，与睾丸的间质区和生精小管区相比，附睾中受体的表达水平更高（分别为18.1±4.4×10²、10.1±2.0×10²和8.3±0.8×10²颗粒/平方微米；P<0.05）。然而，睾丸的间质区和生精小管区之间未检测到受体表达水平的差异。使用针对大鼠催乳素受体胞外域肽序列产生的抗催乳素受体抗体进行免疫细胞化学分析，将受体基因的表达定位到睾丸间质细胞、粗线期精母细胞、圆形精子细胞和伸长精子细胞。在附睾中，受体定位于附睾管内的上皮层。马鹿睾丸和附睾中催乳素受体基因的表达表明该激素在类固醇生成和精子发生中起作用。
