HISTOLOGICAL AND ULTRASTRUCTURAL-LOCALIZATION OF THE KAINATE RECEPTOR SUBUNITS, KA2 AND GLUR6/7, IN THE RAT NERVOUS-SYSTEM USING SELECTIVE ANTIPEPTIDE ANTIBODIES
HISTOLOGICAL AND ULTRASTRUCTURAL-LOCALIZATION OF THE KAINATE RECEPTOR SUBUNITS, KA2 AND GLUR6/7, IN THE RAT NERVOUS-SYSTEM USING SELECTIVE ANTIPEPTIDE ANTIBODIES
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DOI：
10.1002/cne.903490107
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发表时间：
1994-11-01
影响因子：
2.5
通讯作者：
WENTHOLD, RJ
中科院分区：
文献类型：
--
作者：
PETRALIA, RS;WANG, YX;WENTHOLD, RJ
海人藻酸受体广泛存在于大脑的许多区域，可能与神经元对谷氨酸和其他兴奋性氨基酸的反应有关。用C末端多肽制备了识别海人酸结合亚基Ka2和GluR6的亲和纯化多克隆抗体。先前的一项研究表明，尽管GluR6的抗体在一定程度上识别GluR7(因此被命名为GluR6/7)，但每种抗体都是其亚基的特异性。其中一种抗体免疫组织化学染色的震颤切片显示，脑内以及颈髓、背根节和前庭神经节以及松果体和垂体腺的许多结构都有轻到中度的染色。在嗅球、大脑皮层、尾壳核和下丘脑中有中等水平的染色，而丘脑的大部分则呈轻度染色。在海马区，CA3区锥体细胞的染色比CA3区锥体细胞的染色更浓密，与GluR6/7抗体相比差异更明显。此外，CA3区透明层的神经毛染色最深。在脑干，一些感觉核、运动核和网状核的染色呈中等至中等密度。抗GluR6/7抗体阳性的丘脑网状核和桥核呈中等密度染色，可能代表其对GluR7的识别。在小脑中，抗Ka2抗体的颗粒层和分子层及抗GluR6/7抗体的分子层呈中度染色，而抗GluR6/7抗体的颗粒层呈中至浓染。在脑外，抗Ka2抗体染色最浓的是脑垂体腺的中间叶。免疫组织化学染色的超微结构定位于海马区、大脑皮层和小脑皮层。典型地，主要染色在突触后密度，与未染色的带有圆形或主要圆形小泡的突触前终末相对，以及相关的树突。光镜下的染色模式与以前的[H-3]红藻氨酸结合和原位杂交研究非常相似。此外，与以前对其他类型谷氨酸受体分布的研究相比，在神经系统中的许多相同的细胞群中发现了Ka2和GluR6/7以及其他各种亚基。(C)1994年Wiley-Liss，Inc.
Kainate receptors are found throughout many regions of the brain and presumably contribute to responses of neurons to glutamate and other excitatory amino acids. Two affinity-purified polyclonal antibodies that recognize the kainate binding subunits, KA2 and GluR6, were made using C-terminus peptides. A previous study demonstrated that each antibody is specific for its subunit, although antibody to GluR6 recognizes GluR7 to some extent (hence the designation GluR6/7). Vibratome sections immunostained with either antibody showed light to moderate staining in many structures in the brain as well as in cervical spinal cord, dorsal root and vestibular ganglia, and pineal and pituitary glands. Moderate levels were seen in the olfactory bulb, cerebral cortex, caudate/putamen, and hypothalamus, whereas much of the thalamus was stained lightly. In the hippocampus, CA3 pyramidal cells were stained more densely than CAI pyramidal cells-the difference more evident with antibody to GluR6/7. In addition, neuropilar staining was densest in the stratum lucidum of the CA3 region. In the brainstem, staining was moderate to moderately dense in a number of sensory, motor, and reticular nuclei. The moderately dense staining in the reticulothalamic nucleus and pontine nuclei with antibody to GluR6/7 may represent its recognition of GluR7. In the cerebellum, staining was moderate in granular and molecular layers with antibody to KA2 and in the molecular layer with antibody to GluR6/7, whereas it was moderately dense to dense in the granular layer with the GluR6/7 antibody. Outside of the brain, densest staining nas seen with antibody to KA2 in the intermediate lobe of the pituitary gland. Ultrastructural localization of immunostaining was examined in the hippocampus, cerebral cortex, and cerebellar cortex. Typically, major staining was in postsynaptic densities apposed by unstained presynaptic terminals with round or mainly round vesicles and in associated dendrites. The light microscope pattern of staining was fairly similar to that of previous [H-3]kainate binding and in situ hybridization studies. In addition, comparison with previous studies on distribution of other types of glutamate receptors indicates that KA2 and GluR6/7 are found with various other subunits in many of the same cell populations throughout the nervous system. (C) 1994 Wiley-liss, Inc.