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大鼠脑皮质和新纹状体中β-肾上腺素能受体样免疫反应性的超微结构定位

Ultrastructural localization of beta-adrenergic receptor-like immunoreactivity in the cortex and neostriatum of rat brain.

作者信息

Aoki C, Joh T H, Pickel V M

机构信息

Department of Neurology, Cornell University Medical College, New York, NY 10021.

出版信息

Brain Res. 1987 Dec 29;437(2):264-82. doi: 10.1016/0006-8993(87)91642-8.

Abstract

We sought to quantitatively examine the processes containing beta-adrenergic receptor-like immunoreactivity (beta-AR-LI) in the cerebral cortex and neostriatum using a previously characterized rabbit antiserum to frog erythrocyte beta-ARs under optimized immunolabeling conditions. Quantitative assessments of the laminar distribution of beta-AR-LI in the cortex was achieved by computer-assisted image analysis of immunoautoradiographs and by quantitative electron microscopic analysis of peroxidase-antiperoxidase (PAP) labeling in aldehyde-fixed sections and unfixed synaptosomes. In the somatosensory and anterior cingulate cortical areas, light microscopy of aldehyde-fixed sections immunolabeled by the PAP method revealed small (0.5-1.0 micron) punctate processes in all layers. In the deeper layers, rims of immunoreactivity around the plasmalemma of a population of neuronal perikarya and processes were also observed. By immunoautoradiography, labeling was seen in distinct, laminar distributions resembling the reported autoradiographic patterns using radioligands. By electron microscopy, the immunoreactive profiles in all cortical layers were primarily thick and thin postsynaptic densities (PSDs), comprising 4% of all identifiable PSDs in fixed sections and 12% in unfixed synaptosomal preparations. Also labeled were saccules of smooth endoplasmic reticulum and pinocytotic vesicles in dendrites, glial processes and lightly myelinated axons. In the neostriatum, the density of autoradiographic immunoreactivity was equivalent to the heavily labeled laminae of the cerebral cortex. Immunoreactivity detectable by light microscopy included punctate processes and rims of perikarya, as was seen in the cerebral cortex. The PAP reaction was shown by electron microscopy to be localized to the cytoplasmic surface of plasmalemma of a few proximal dendrites, but was most prominently associated with PSDs of dendritic spines. Preadsorption of the antiserum with a partially purified beta-AR preparation abolished all detectable immunoreactivity. These results provide further support for the specificity of the antiserum for beta-ARs, and are the first quantitative ultrastructural evidence for association of beta-AR-LI with PSDs in the cerebral cortex. The neostriatum, whose major catecholaminergic innervation is dopaminergic, and not noradrenergic, is also confirmed to exhibit high levels of beta-AR-LI within subcellular structures analogous to those seen in the cerebral cortex.(ABSTRACT TRUNCATED AT 400 WORDS)

摘要

我们试图在优化的免疫标记条件下,使用先前鉴定的针对蛙红细胞β-肾上腺素能受体(β-AR)的兔抗血清,对大脑皮质和新纹状体中含有β-肾上腺素能受体样免疫反应性(β-AR-LI)的过程进行定量检测。通过对免疫自显影片的计算机辅助图像分析,以及对醛固定切片和未固定突触小体中过氧化物酶抗过氧化物酶(PAP)标记的定量电子显微镜分析,实现了对皮质中β-AR-LI层状分布的定量评估。在体感和前扣带回皮质区域,用PAP法免疫标记的醛固定切片的光学显微镜检查显示,所有层中均有小的(0.5-1.0微米)点状突起。在较深的层中,还观察到一群神经元胞体和突起的质膜周围有免疫反应性边缘。通过免疫自显影,可见标记呈明显的层状分布,类似于使用放射性配体报道的自显影模式。通过电子显微镜观察,所有皮质层中的免疫反应性轮廓主要是厚和薄的突触后致密物(PSD),在固定切片中占所有可识别PSD的4%,在未固定的突触小体制剂中占12%。树突、神经胶质突起和轻度髓鞘化轴突中的滑面内质网囊泡和吞饮小泡也有标记。在新纹状体中,放射自显影免疫反应性的密度与大脑皮质标记密集的层相当。光学显微镜可检测到的免疫反应性包括点状突起和胞体边缘,与大脑皮质中所见相同。电子显微镜显示,PAP反应定位于少数近端树突质膜的细胞质表面,但最显著地与树突棘的PSD相关。用部分纯化的β-AR制剂对抗体进行预吸附可消除所有可检测到的免疫反应性。这些结果进一步支持了该抗血清对β-AR的特异性,并且是β-AR-LI与大脑皮质中PSD相关联的首个定量超微结构证据。主要儿茶酚胺能神经支配为多巴胺能而非去甲肾上腺素能的新纹状体,也被证实在类似于大脑皮质所见的亚细胞结构中表现出高水平的β-AR-LI。(摘要截短至400字)

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