PROLACTIN RECEPTOR MESSENGER-RNA LOCALIZATION IN THE HYPOTHALAMUS BY IN-SITU HYBRIDIZATION
PROLACTIN RECEPTOR MESSENGER-RNA LOCALIZATION IN THE HYPOTHALAMUS BY IN-SITU HYBRIDIZATION
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DOI：
10.1111/j.1365-2826.1994.tb00572.x
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发表时间：
1994-04-01
影响因子：
3.2
通讯作者：
WISE, PM
中科院分区：
文献类型：
--
作者：
CHIU, SF;WISE, PM
催乳素受体可介导催乳素在脑中的作用，从而影响行为和神经内分泌。我们最近证明催乳素受体基因表达的前和内侧基底下丘脑，而不是在皮质的逆转录聚合酶链反应。在本文中，我们定位催乳素受体基因表达的单个细胞与原位杂交。通过使用（i）与长和短催乳素受体共同的细胞外结合结构域的编码序列互补的探针，（ii）更严格的杂交和洗涤条件以减少背景，以及（iii）更高比活性、更复杂和饱和量的探针，对原位杂交方法中的几个步骤进行了修改，以提高灵敏度。我们检测了催乳素受体基因在视前核室周区、视前内侧核、视上核、弓状核吻端和脉络丛细胞中的表达。皮质脑组织，这已被证明是以前通过逆转录聚合酶链反应缺乏催乳素受体mRNA，没有任何可检测的信号的受体mRNA，并被用作信号的背景水平的指示。在视前核、内侧视前核、视上核、弓状核、外侧腹内侧核的室周区，标记细胞上的银颗粒平均面积至少是同一脑切片皮质背景的10倍。
Prolactin receptors may mediate the action of prolactin in the brain to influence behavior and neuroendocrine secretions. We recently demonstrated prolactin receptor gene expression in the anterior and medial basal hypothalamus and not in the cortex by the reverse transcription-polymerase chain reaction. In this paper, we localize the prolactin receptor gene expression to individual cells with in situ hybridization. Several steps in the in situ hybridization method were modified to increase sensitivity by using (i) probes complementary to the coding sequence of the extracellular binding domain common to both long and short prolactin receptor, (ii) more stringent hybridization and wash conditions to reduce background and (iii) higher specific activity, more complex and saturating amounts of probe. We detected prolactin receptor gene expression in cells of the periventricular area of the preoptic nucleus, medial preoptic nucleus, supraoptic nucleus, rostral arcuate nucleus and choroid plexus. Cortical brain tissue, which has been demonstrated previously by reverse-transcription polymerase chain reaction to be lacking in prolactin receptor mRNA, did not have any detectable signal for the receptor mRNA and was used as an indication of background levels of signal. The mean area of silver grains over labeled cells in periventricular area of the preoptic nucleus, medial preoptic nucleus, supraoptic nucleus, arcuate nucleus, lateral ventromedial nucleus was at least 10 times greater than the background in the cortex of the same brain section.