Subcellular localization of calcium-permeable AMPA receptors in spinal motoneurons.

Subcellular localization of calcium-permeable AMPA receptors in spinal motoneurons.
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脊髓运动神经元中钙渗透性 AMPA 受体的亚细胞定位。

DOI:
10.1046/j.0953-816x.2001.01648.x
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发表时间:
2001
期刊:
The European journal of neuroscience
影响因子:
--
通讯作者:
Brorson,JR
Brorson,JR
中科院分区:
--
文献类型:
--
作者:
Vandenberghe,W;Bindokas,VP;Miller,RJ;Robberecht,W;Brorson,JR

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钙离子通透性α-氨基-3-羟基-5-甲基-4-异恶唑丙酸受体的激活与脊髓运动神经元的存活和树突状突起的生长密切相关。AMPA受体的Ca~(2+)通透性受GluR2亚基控制。全细胞电生理学研究表明,单个运动神经元中可能同时存在含GluR2和不含GluR2的AMPA受体。然而,还没有直接证明单个运动神经元中AMPA受体亚单位组成的异质性,也没有明显的亚细胞分布的含GluR2和缺乏GluR2的受体。在本研究中,我们利用共聚焦显微镜、免疫细胞化学和钙离子成像技术对培养的大鼠脊髓运动神经元内AMPA受体的亚细胞定位进行了研究。GluR2和GluR4的免疫反应主要集中在簇状结构中,绝大多数分布在突触的树突中。GluR2和GluR4的双重标记显示,GluR2和GluR4的相对表达在单个运动神经元内的簇之间存在差异;大多数AMPA受体簇同时表达GluR2和GluR4,但相当少数的簇仅表达GluR2或仅表达GluR4。GluR2免疫阴性的AMPA受体簇主要存在于树突中,但GluR2免疫阴性簇和GluR2阳性簇在树突和胞体中的相对比例相似。AMPA受体激活触发的[Ca~(2+)]虹膜成像证实,Ca~(2+)流入运动神经元树突。这些发现有力地支持了一种模型,即含GluR2和不含GluR2的AMPA受体共存于运动神经元中,聚集在突触上,并以相对比例混合,在细胞膜微域之间存在很大差异。
Activation of Ca2+‐permeable α‐amino‐3‐hydroxy‐5‐methyl‐4‐isoxazole propionic acid (AMPA) receptors has been linked to potent effects on survival and dendritic outgrowth of spinal motoneurons. Ca2+permeability of AMPA receptors is controlled by the GluR2 subunit. Whole‐cell electrophysiological studies have suggested that GluR2‐containing and GluR2‐lacking AMPA receptors may coexist in individual motoneurons. However, there has not been a direct demonstration of heterogeneity in AMPA receptor subunit composition in single motoneurons, nor of distinct subcellular distributions of GluR2‐containing and GluR2‐lacking receptors. In the present study, we have used confocal microscopy, immunocytochemistry and Ca2+imaging to characterize the subcellular localization of AMPA receptors in cultured rat spinal motoneurons. Immunoreactivity for GluR2 and GluR4 was concentrated in clusters, the vast majority of which were found in dendrites at synapses. Double‐labelling for GluR2 and GluR4 revealed variability in relative expression of GluR2 and GluR4 between clusters within individual motoneurons; most AMPA receptor clusters were immunoreactive for both GluR2 and GluR4, but a significant minority of clusters were immunoreactive for GluR2 only or for GluR4 only. The majority of GluR2‐immunonegative AMPA receptor clusters was present in dendrites, but the relative proportion of GluR2‐immunonegative and GluR2‐immunopositive clusters was similar in dendrites and soma. Imaging of [Ca2+]irises triggered by AMPA receptor activation confirmed Ca2+influx in motoneuron dendrites. These findings strongly support a model in which GluR2‐containing and GluR2‐lacking AMPA receptors coexist in motoneurons, clustered at synapses, and mixed in a relative proportion that varies considerably between cell membrane microdomains.
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