Crumeyrolle-Arias M, Latouche J, Jammes H, Djiane J, Kelly P A, Reymond M J, Haour F
CNRS-Institut Pasteur, Paris, France.
Neuroendocrinology. 1993 Mar;57(3):457-66. doi: 10.1159/000126392.
A precise mapping of prolactin (PRL) receptors in the rat brain has been achieved. Localization of binding sites for both 125I-human growth hormone (125I-hGH) and 125I-monoclonal anti-PRL receptor (125I-U5) was studied by in vitro autoradiography on brain sections in female rats (n = 7). The analysis of autoradiograms generated from 12 adjacent sections at 11 different brain levels (bregma 0.2 to -4.8 mm) revealed 9 distinctive localizations for 125I-hGH binding sites: preoptic suprachiasmatic nucleus, medial preoptic area, periventricular, supraoptic, paraventricular, arcuate and vetromedial nuclei and also the median eminence and the infundibulum. Specificity for PRL binding was assessed by competition experiment of 125I-hGH with unlabeled hGH and ovine PRL. Binding sites were similarly localized by 125I-U5 indicating the presence of PRL receptors moiety. The quantitative analysis with 0.6 nM 125I-hGH demonstrated maximal densities in the preoptic suprachiasmatic and arcuate nuclei and minimal densities in the median eminence and the infundibulum. Due to ample antero-posterior variations no significant changes were observed during the estrous cycle. Saturation analysis of binding in the arcuate nucleus indicated a single class of high affinity (Kd from 0.9 to 2.2 nM) receptors (Bmax from 34 to 44 fmol/mg of proteins). The present data provide the hypothalamic cartography of PRL receptors in the female rat brain and support all the physiological evidence for the existence of a direct action of PRL in the hypothalamus.
已在大鼠脑中实现了催乳素(PRL)受体的精确图谱绘制。通过对雌性大鼠(n = 7)脑切片进行体外放射自显影,研究了¹²⁵I-人生长激素(¹²⁵I-hGH)和¹²⁵I-单克隆抗PRL受体(¹²⁵I-U5)的结合位点定位。对来自11个不同脑水平(前囟0.2至-4.8毫米)的12个相邻切片生成的放射自显影片分析显示,¹²⁵I-hGH结合位点有9个独特的定位:视前交叉上核、内侧视前区、室周区、视上核、室旁核、弓状核和腹内侧核,以及正中隆起和漏斗部。通过¹²⁵I-hGH与未标记的hGH和羊PRL的竞争实验评估了PRL结合的特异性。¹²⁵I-U5同样定位了结合位点,表明存在PRL受体部分。用0.6 nM ¹²⁵I-hGH进行的定量分析表明,视前交叉上核和弓状核中的密度最大,而正中隆起和漏斗部中的密度最小。由于前后变化很大,在发情周期中未观察到明显变化。弓状核结合的饱和分析表明存在一类高亲和力(Kd为0.9至2.2 nM)受体(Bmax为34至44 fmol/mg蛋白质)。目前的数据提供了雌性大鼠脑中PRL受体的下丘脑图谱,并支持了PRL在下丘脑中存在直接作用的所有生理学证据。