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目的探讨大鼠接受高渗刺激后,视上核和孤束核内神经元-星形胶质细胞复合体(N-ASC)反应。方法成年雄性SD大鼠50只,随机分为光镜组(n=40)和电镜组(n=10)。采用多重免疫荧光标记法,在光镜下观察高渗刺激(9%氯化钠,5.5ml/kg尾静脉注射)后15、45、90和180 min,视上核内胶质纤维酸性蛋白(GFAP,标记星形胶质细胞)、催产素(OXT,标记视上核神经元)、Fos(标记神经元胞核)和孤束核内GFAP、酪胺酸羟化酶(TH,标记神经元)、Fos表达的时程变化。采用双重免疫电镜法,观察高渗刺激后视上核N-ASC内星形胶质细胞(用Cx43标记)与神经元(用Cx32标记)之间接触处的超微结构。结果光镜下在视上核和孤束核可分别观察到3种N-ASC,即:由GFAP阳性星形胶质细胞与OXT(或TH)阳性神经元形成的N-ASC;由GFAP阳性星形胶质细胞与Fos阳性神经元形成的N-ASC;由GFAP阳性星形胶质细胞与Fos和OXT(或TH)双阳性神经元形成的N-ASC。高渗刺激后N-ASC的数量明显增加。抗Cx43和抗Cx32的双重免疫电镜显示,高渗刺激后Cx43阳性星形胶质细胞突起(即Cx43半通道)以及Cx32和Cx43形成的异型缝隙连接(HGJ)的数量明显增加。结论视上核和孤束核内的N-ASC参与渗透压调节反应。
Objective To investigate the neuronal-astrocyte complex (N-ASC) response in supraoptic nucleus and nucleus tractus solitarius in rats after hypertonic stimulation. Methods 50 male SD rats were randomly divided into light microscope group (n = 40) and electron microscope group (n = 10). Multiple immunofluorescent labeling method was used to observe the effect of hypertonic stimulation (9% sodium chloride, 5.5ml / kg tail vein injection) at 15, 45, 90 and 180 minutes after light injection. The glial fibrillary acidic protein GFAP, labeled astrocytes), oxytocin (OXT, labeled supraoptic neurons), Fos (labeled neuronal nuclei) and nucleus tractus solitarius GFAP, tyrosine hydroxylase (TH, labeled neurons ), The time course of Fos expression. Double immunoelectron microscopy was used to observe the ultrastructure of contact between neurons (labeled with Cx32) in astrocytes (labeled with Cx43) in supraoptic nucleus N-ASC after hypertonic stimulation. Results Three types of N-ASC were observed in the supraoptic nucleus and nucleus tractus solitarium under light microscope: N-ASC formed by GFAP positive astrocytes and OXT (or TH) positive neurons, N-ASC formed by astrocytes and Fos-positive neurons; N-ASC formed by GFAP-positive astrocytes and Fos and OXT (or TH) double-positive neurons. The number of N-ASC increased significantly after hypertonic stimulation. Dual immunoelectron microscopy of anti-Cx43 and anti-Cx32 showed a significant increase in the number of Cx43-positive astrocyte protrusions (ie, Cx43 hemichannels) and shaped gap junctions (HGJs) formed by Cx32 and Cx43 after hypertonic stimulation. Conclusion N-ASC in the supraoptic nucleus and nucleus tractus solitarii participate in the regulation of osmotic pressure.