Shirota M, Kurohmaru M, Hayashi Y, Shirota K, Kelly P A
Laboratory of Reproductive and Developmental Toxicology, Hatano Research Institute of Food and Drug Safety Center, Kanagawa, Japan.
Endocr J. 1995 Feb;42(1):69-76. doi: 10.1507/endocrj.42.69.
Biotinylated riboprobe that specifically hybridized with messenger RNAs (mRNAs) encoding the long form of prolactin receptor (PRLRL) was transcribed from 269 bp Hind III and Xho I fragment of the cytoplasmic domain of PRLRL complementary DNA (cDNA). The probe was used for in situ hybridization to identify tissue localization of PRLRL mRNA in the mammary gland, liver, ovaries and kidneys from lactating rats at 24-36 h after delivery. Histochemical detection of signals by means of horseradish peroxidase (HRP)-labeled streptavidin revealed that the PRLRL gene was expressed on the alveolar epithelial cells and mammary ductal epithelium in the mammary gland, and hepatocytes in the liver. In the ovary, the PRLRL gene was expressed on the luteal cells in the newly formed corpus luteum, granulosa cells and theca cells of follicles at various stages of development, hypertrophied theca cells in the atretic follicle, and secondary interstitial cells, but no signal was observed in the kidney. Biotinylated sense RNA probe did not detect any signals in any of the tissues examined. In situ hybridization with non-radiolabeled probe provided the identification of PRLRL mRNA in the fine tissues, such as follicular epithelium in the ovary, and showed the morphology of individual cells expressing the PRLRL gene. In particular, the diversity of signal intensity in the same mammary gland with different appearances suggested the existence of a local mechanism controlling PRLRL gene expression.
生物素化核糖探针与编码催乳素受体长型（PRLRL）的信使核糖核酸（mRNA）特异性杂交，该探针由PRLRL互补脱氧核糖核酸（cDNA）胞质结构域的269 bp Hind III和Xho I片段转录而成。该探针用于原位杂交，以鉴定分娩后24 - 36小时泌乳大鼠乳腺、肝脏、卵巢和肾脏中PRLRL mRNA的组织定位。通过辣根过氧化物酶（HRP）标记的链霉亲和素进行信号的组织化学检测显示，PRLRL基因在乳腺的肺泡上皮细胞和乳腺导管上皮以及肝脏的肝细胞中表达。在卵巢中，PRLRL基因在新形成的黄体中的黄体细胞、不同发育阶段卵泡的颗粒细胞和卵泡膜细胞、闭锁卵泡中肥大的卵泡膜细胞以及次级间质细胞中表达，但在肾脏中未观察到信号。生物素化的正义RNA探针在任何检测的组织中均未检测到任何信号。与非放射性标记探针的原位杂交确定了PRLRL mRNA在精细组织中的位置，如卵巢中的卵泡上皮，并显示了表达PRLRL基因的单个细胞的形态。特别是，同一乳腺中不同外观的信号强度差异表明存在控制PRLRL基因表达的局部机制。
