Silverman W F, Walsh R J, Posner B I
Brain Res. 1986 Jan;389(1-2):11-9. doi: 10.1016/0165-3806(86)90168-9.
The development of prolactin receptors in the choroid plexus of the rat was examined using the in vivo autoradiographic approach employing the principle of competitive binding. Experimental animals were injected with [125I]prolactin alone (total binding) while control animals received [125I]prolactin and a 500-fold excess of unlabelled prolactin (non-specific binding). Newborns as well as animals 10, 14 and 18 days postnatal were studied. Three minutes following hormone injection animals received an intracardiac perfusion with fixative and tissues were prepared for quantitative light microscopic autoradiography. The choroid plexus first demonstrated specific binding of prolactin, i.e. a statistically significant difference in the autoradiographic reactions between experimental and control animals, at 14 days postnatal. The lactogen specificity of these binding sites was further defined by the ability of [125I]prolactin to be displaced by unlabelled human growth hormone, which is lactogenic in rats, and not by unlabelled insulin, which is structurally dissimilar to prolactin. Morphometric analyses were performed on electron micrographs of choroid plexus from 10- and 14-day postnatal rats. The volume densities of constituents known to be enriched in polypeptide hormone receptors were measured and compared. Small cytoplasmic vesicles and tubules were statistically significantly more abundant in 10-day-old rats than in 14-day-old animals. It is conjectured that these vesicles and tubules contain an intracellular pool of prolactin receptors whose decrease at 14 days parallels the expression of specific binding sites at the cell surface.
采用竞争结合原理的体内放射自显影方法,研究了大鼠脉络丛中催乳素受体的发育情况。给实验动物单独注射[125I]催乳素(总结合),而对照动物接受[125I]催乳素和500倍过量的未标记催乳素(非特异性结合)。对新生大鼠以及出生后10天、14天和18天的动物进行了研究。激素注射3分钟后,给动物进行心脏内灌注固定剂,并制备组织用于定量光学显微镜放射自显影。脉络丛在出生后14天首次表现出催乳素的特异性结合,即实验动物和对照动物在放射自显影反应上存在统计学上的显著差异。这些结合位点的催乳素特异性通过未标记的人生长激素(在大鼠中具有催乳素活性)能够取代[125I]催乳素,而未标记的胰岛素(其结构与催乳素不同)不能取代[125I]催乳素的能力进一步得到定义。对出生后10天和14天大鼠脉络丛的电子显微镜照片进行了形态计量分析。测量并比较了已知富含多肽激素受体的成分的体积密度。在10日龄大鼠中,小细胞质囊泡和小管在统计学上比14日龄动物丰富得多。据推测,这些囊泡和小管含有催乳素受体的细胞内池,其在14天时的减少与细胞表面特异性结合位点的表达平行。