Localization of neurones expressing the gap junction protein Connexin45 within the adult spinal dorsal horn: a study using Cx45-eGFP reporter mice.
Localization of neurones expressing the gap junction protein Connexin45 within the adult spinal dorsal horn: a study using Cx45-eGFP reporter mice.
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DOI：
10.1007/s00429-012-0426-1
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发表时间：
2013-05
影响因子：
3.1
通讯作者：
King, A. E.
中科院分区：
文献类型：
--
作者：
Chapman, R. J.;Lall, V. K.;Maxeiner, S.;Willecke, K.;Deuchars, J.;King, A. E.
定位于神经元和神经胶质合胞体的连接蛋白 (Cx) 为通过间隙连接的细胞间通讯提供超微结构成分。在本研究中，使用 Cx45 报告小鼠模型（其中 Cx45 编码序列替换了增强型绿色荧光蛋白 (eGFP)）来表征成年小鼠脊髓内表达 Cx45 的神经元。 eGFP 免疫反应性 (eGFP-IR) 细胞定位于背角 (DH) 的 I-III 层的所有头尾水平，即与伤害感受相关的区域。通过 eGFP-IR 与神经元标记 NeuN 的共定位证实了 DH 中这些细胞的神经元表型而不是神经胶质表型。进一步的免疫组织化学研究表明，eGFP-IR 中间神经元共表达钙结合蛋白钙结合蛋白，并在较小程度上表达钙结合蛋白。相比之下，eGFP-IR 谱不与小白蛋白或 GAD-67 共定位，这两者都与抑制性中间神经元相关。用主要传入标记物异凝集素-B4 (IB4) 和降钙素基因相关肽染色显示，I-III 层内的 eGFP-IR 体体与先前假定的外周起源的非肽能伤害性传入细胞紧密相连。 5-HT 末端与 eGFP-IR 神经元间体紧密相连的存在表明通过下行通路进行调节。这些数据表明 Cx45 在小鼠浅表背角内的中间神经元群体中高度局部化表达。讨论了这些数据在 Cx45 和间隙连接在脊髓体感处理和疼痛中的假定作用的背景下的含义。
Connexin (Cx) proteins localized to neuronal and glial syncytia provide the ultrastructural components for intercellular communication via gap junctions. In this study, a Cx45 reporter mouse model in which the Cx45 coding sequence is substituted for enhanced green fluorescent protein (eGFP) was used to characterize Cx45 expressing neurones within adult mouse spinal cord. eGFP-immunoreactive (eGFP-IR) cells were localized at all rostro-caudal levels to laminae I–III of the dorsal horn (DH), areas associated with nociception. The neuronal rather than glial phenotype of these cells in DH was confirmed by co-localisation of eGFP-IR with the neuronal marker NeuN. Further immunohistochemical studies revealed that eGFP-IR interneurones co-express the calcium-binding protein calbindin, and to a lesser extent calretinin. In contrast, eGFP-IR profiles did not co-localize with either parvalbumin or GAD-67, both of which are linked to inhibitory interneurones. Staining with the primary afferent markers isolectin-B4 (IB4) and calcitonin gene-related peptide revealed that eGFP-IR somata within laminae I–III receive close appositions from the former, presumed non-peptidergic nociceptive afferents of peripheral origin. The presence of 5-HT terminals in close apposition to eGFP-IR interneuronal somata suggests modulation via descending pathways. These data demonstrate a highly localized expression of Cx45 in a population of interneurones within the mouse superficial dorsal horn. The implications of these data in the context of the putative role of Cx45 and gap junctions in spinal somatosensory processing and pain are discussed.
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影响因子：
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