Pérez Armendariz E Martha, Norcini Monica, Hernández-Tellez Beatriz, Castell-Rodríguez Andrés, Coronel-Cruz Cristina, Alquicira Raquel Guerrero, Sideris Alexandra, Recio-Pinto Esperanza
Departamento de Biología Celular y Tisular, Facultad de Medicina, Universidad Nacional Autónoma de México, Torre de Investigación 5to piso, Avenida Universidad 3000, Circuito Interior, Ciudad Universitaria, Colonia Universidad Nacional Autónoma de México, CU, D.F., 04510, Mexico.
Department of Anesthesiology, NYULMC, 180 Varick Street, Room 677, New York, NY 10014, USA.
Acta Histochem. 2018 Apr;120(3):168-178. doi: 10.1016/j.acthis.2017.11.005. Epub 2017 Dec 8.
Previous studies have shown that following peripheral nerve injury there was a downregulation of the gap junction protein connexin 36 (Cx36) in the spinal cord; however, it is not known whether Cx36 protein is expressed in the dorsal root ganglia (DRGs), nor if its levels are altered following peripheral nerve injuries. Here we address these aspects in the adult rat lumbar DRG. Cx36 mRNA was detected using qRT-PCR, and Cx36 protein was identified in DRG sections using immunohistochemistry (IHC) and immunofluorescence (IF). Double staining revealed that Cx36 co-localizes with both anti-β-III tubulin, a neuronal marker, and anti-glutamine synthetase, a satellite glial cell (SGC) marker. In neurons, Cx36 staining was mostly uniform in somata and fibers of all sizes and its intensity increased at the cell membranes. This labeling pattern was in contrast with Cx36 IF dots mainly found at junctional membranes in islet beta cells used as a control tissue. Co-staining with anti-Cx43 and anti-Cx36 showed that whereas mostly uniform staining of Cx36 was found throughout neurons and SGCs, Cx43 IF puncta were localized to SGCs. Cx36 mRNA was expressed in normal lumbar DRG, and it was significantly down-regulated in L4 DRG of rats that underwent sciatic nerve injury resulting in persistent hypersensitivity. Collectively, these findings demonstrated that neurons and SGCs express Cx36 protein in normal DRG, and suggested that perturbation of Cx36 levels may contribute to chronic neuropathic pain resulting from a peripheral nerve injury.
先前的研究表明,周围神经损伤后脊髓中缝隙连接蛋白连接蛋白36(Cx36)表达下调;然而,尚不清楚Cx36蛋白是否在背根神经节(DRG)中表达,也不清楚其水平在周围神经损伤后是否发生改变。在此,我们在成年大鼠腰段DRG中研究这些方面。使用qRT-PCR检测Cx36 mRNA,并使用免疫组织化学(IHC)和免疫荧光(IF)在DRG切片中鉴定Cx36蛋白。双重染色显示,Cx36与神经元标志物抗β-III微管蛋白和卫星神经胶质细胞(SGC)标志物抗谷氨酰胺合成酶均共定位。在神经元中,Cx36染色在所有大小的胞体和纤维中大多均匀,其强度在细胞膜处增加。这种标记模式与用作对照组织的胰岛β细胞中主要在连接膜处发现的Cx36 IF点形成对比。抗Cx43和抗Cx36的共染色显示,虽然在整个神经元和SGC中发现Cx36大多呈均匀染色,但Cx43 IF斑点定位于SGC。Cx36 mRNA在正常腰段DRG中表达,在坐骨神经损伤导致持续超敏反应的大鼠L4 DRG中显著下调。总的来说,这些发现表明神经元和SGC在正常DRG中表达Cx36蛋白,并提示Cx36水平的扰动可能导致周围神经损伤引起的慢性神经性疼痛。