Inagaki S, Kito S, Kubota Y, Girgis S, Hillyard C J, MacIntyre I
Brain Res. 1986 May 28;374(2):287-98. doi: 10.1016/0006-8993(86)90423-3.
125I-calcitonin gene-related peptide (CGRP) binding sites were mapped in the human brain and rat brains by in vitro macroautoradiography, and compared to each other. Binding experiments were made to characterize 125I-CGRP binding on the human and rat brains. Scatchard analysis of saturation experiments from slide-mounted sections of the human and rat cerebellum displayed 125I-CGRP binding sites with a dissociation constant (Kd) of 0.17 nM and 0.11 nM, respectively, and a maximal number of binding sites (Bmax) of 96.8 fmol/mg and 23.0 fmol/mg protein. 125I-CGRP binding was time-dependent, reversible and saturable with high affinity in the brains. Autoradiograms showed a discrete distribution of 125I-CGRP binding sites throughout the brains of human and rat with patterns similar to each other. In the human brain, the highest binding was seen in the cerebellum, inferior olivary nuclear complex, certain parts of the central gray matter, arcuate nuclei of the medulla oblongata and dorsal motor nucleus of the vagus, and densities of CGRP-binding sites were high in the nucleus accumbens, amygdala, tail of the nucleus caudatus, substantia nigra, ventral tegmental area, medial portion of the inferior colliculus, medial pontine nuclei, locus coeruleus, inferior vestibular nucleus, substantia gelatinosa of the spinal trigeminal nucleus, nucleus of the solitary tract and nucleus cuneatus lateralis. In the rat, high densities were found in the hippocampus pars anterior, nucleus accumbens, ventral and caudal portions of the nucleus caudatus-putamen, central and basolateral nuclei of the amygdala, caudal portion of the insular cortex, medial geniculate body, superior and inferior colliculi, certain portions of the central gray matter, locus coeruleus, inferior olivary nuclei, vagal complex, nucleus cuneatus lateralis and cerebellum. In contrast, in both species, most of the cortical areas including the hippocampus, most of the thalamus, and hypothalamus exhibited few binding sites. In addition, high quantities of the binding sites were seen on the pia mater and on walls of blood vessels in the brain and subarachnoidea. These results revealed essentially homologous locations of CGRP binding sites in the human and rat central nervous systems and well corresponding distributions of binding sites and endogenous CGRP-like immunoreactivity.
通过体外宏观放射自显影技术在人脑和大鼠脑中绘制了125I-降钙素基因相关肽(CGRP)结合位点,并进行了相互比较。进行结合实验以表征125I-CGRP在人脑和大鼠脑中的结合情况。对人及大鼠小脑载玻片切片的饱和实验进行Scatchard分析显示,125I-CGRP结合位点的解离常数(Kd)分别为0.17 nM和0.11 nM,最大结合位点数(Bmax)分别为96.8 fmol/mg和23.0 fmol/mg蛋白质。125I-CGRP在脑中的结合具有时间依赖性、可逆性且具有高亲和力的饱和性。放射自显影片显示,在人和大鼠的脑中,125I-CGRP结合位点呈离散分布,模式相似。在人脑中,小脑、下橄榄核复合体、中央灰质的某些部分、延髓弓状核和迷走神经背运动核的结合最高,伏隔核、杏仁核、尾状核尾部、黑质、腹侧被盖区、下丘内侧部分、脑桥内侧核、蓝斑、下前庭核、三叉神经脊束核胶状质、孤束核和外侧楔束核的CGRP结合位点密度较高。在大鼠中,海马前部、伏隔核、尾壳核的腹侧和尾部、杏仁核的中央和基底外侧核、岛叶皮质尾部、内侧膝状体、上丘和下丘、中央灰质的某些部分、蓝斑、下橄榄核、迷走神经复合体、外侧楔束核和小脑的密度较高。相比之下,在这两个物种中,包括海马在内的大多数皮质区域、大多数丘脑和下丘脑的结合位点很少。此外,在软脑膜以及脑和蛛网膜下腔的血管壁上可见大量结合位点。这些结果揭示了CGRP结合位点在人和大鼠中枢神经系统中的基本同源位置,以及结合位点与内源性CGRP样免疫反应性的良好对应分布。