Ouhtit A, Morel G, Kelly P A
CNRS URA 1454, Neuroendocrinologie, Faculté de Médecine Lyon-Sud, Oullins, France.
Endocrinology. 1993 Jul;133(1):135-44. doi: 10.1210/endo.133.1.8319561.
PRL receptor gene expression was visualized in various tissues by in situ hybridization, using 35S-labeled probes unique to each form of receptor. Tissues were removed rapidly from adult male and female rats and placed in liquid nitrogen. Cryostat sections (10 microns) were prepared, fixed, pretreated, and dehydrated before incubation with the various probes. Hybridization was performed overnight, after which the slides were first exposed to autoradiographic film and then dipped in nuclear emulsion and exposed for 1-2 weeks. The specificity of the signal was studied by competition and using radiolabeled heterologous probes. Some tissues show no expression of either form of receptor mRNA, such as olfactory bulb and penis. Macroautoradiogram signals (optical density) were compared to a standard curve to observe the variation in mRNA expression, which was expressed in arbitrary units. Sex differences in the expression of PRL receptors were seen in a number of tissues, such as adrenal gland and pituitary. Expression of mRNAs specific to the long form of PRL receptor was predominant in adrenal gland, pituitary, thymus, spleen, skin, heart, and skeletal muscle, whereas the short form was expressed to a greater extent in kidney and lung. At the light microscopic level, the silver grains observed by epipolarization or light field were seen in the specific regions or cells that express PRL receptor mRNAs. In conclusion, the long form transcript predominates in most tissues, except kidney and lung. The advantage of in situ hybridization is that it allows the identification of specific regions or cells expressing mRNAs to be identified. The actions of PRL have not been identified in all tissues expressing PRL receptor transcripts, nor has any definitive correlation been made with the expression of short and long forms of PRL receptor and function.
使用针对每种受体形式的35S标记探针，通过原位杂交技术在各种组织中观察催乳素（PRL）受体基因的表达情况。从成年雄性和雌性大鼠中迅速取出组织并置于液氮中。制备（10微米厚的）冰冻切片，固定、预处理并脱水，然后与各种探针一起孵育。杂交进行过夜，之后玻片先与放射自显影片接触，然后浸入核乳胶中并曝光1 - 2周。通过竞争实验和使用放射性标记的异源探针研究信号的特异性。一些组织未显示任何一种受体mRNA的表达，如嗅球和阴茎。将宏观放射自显影片信号（光密度）与标准曲线进行比较，以观察mRNA表达的变化，其以任意单位表示。在许多组织中，如肾上腺和垂体，观察到PRL受体表达存在性别差异。PRL受体长形式特异性mRNA在肾上腺、垂体、胸腺、脾脏、皮肤、心脏和骨骼肌中表达占主导，而短形式在肾脏和肺中表达程度更高。在光学显微镜水平，通过偏光或明场观察到的银颗粒出现在表达PRL受体mRNA的特定区域或细胞中。总之，除肾脏和肺外，长形式转录本在大多数组织中占主导。原位杂交的优点在于它能够识别表达mRNA的特定区域或细胞。PRL的作用在所有表达PRL受体转录本的组织中尚未明确，PRL受体长短形式的表达与功能之间也未建立任何明确的相关性。
