成年大鼠纹状体nNOS阳性神经元的形态特征及其对DA剥夺的反应

The morphology of striatal nNOS neurons and response to DA depletion in adult rats

目的探讨成年雄性大鼠纹状体nNOS中间神经元对6OHDA诱导性DA剥夺的反应。方法选取60只成年雄性SD大鼠,随机分成正常对照组、生理盐水注射组、6OHDA处理组。实验借助6OHDA模型、免疫组化、免疫荧光、免疫印迹技术探察证实纹状体n NOS阳性中间神经元对DA剥夺的反应,实验数据用SPSS 20.0统计软件进行分析。结果免疫荧光双标记结果显示,nNOS阳性中间神经元与NPY神经元存在极高的共存率(98.5±0.1)%,其与C AT阳性中间神经元也存在较低的共存率(2.4±2.0)%。nNOS阳性神经元与Cr和Parv阳性中间神经元之间未观察到共存关系。免疫组化资料显示,n NOS阳性神经元胞体呈现梭形、圆形,nNOS胞体大小和主树突数量在对照组[分别为(13.4±1.3)μm、2.00±0.94)与6OHDA组[分别为(13.4±1.8)μm、1.77±0.73)之间差异无统计学意义(P≥0.05)。但6OHDA处理组n NOS阳性神经元胞体的数量(1.48±0.19)%显著多于对照组(0.77±0.13)%(P<0.05)。Western blot技术探测显示,6OHDA组(0.98±0.07)的n NOS蛋白量表达量也显著高于对照组(0.56±0.04;P<0.05)。结论 6OHDA诱导性DA剥夺明显导致纹状体n NOS阳性神经元的特征性反应及其蛋白高表达,此提示n NOS阳性神经元可能牵涉到DA剥夺诱导纹状体损伤的病理过程。

Objective nNOS-positive neurons that play an important regulatory role in the central nervous system, was involved in kinds of diseases, pathological processes. This study is to furtherly explore the response of nNOS neurons to DA depletion. Methods 60 SD male adult rats （250-350 g） were selected and randomly divided into three groups： blank control group, 0.9% NaC1 injection group and 6OHDA group. We use immunofluorescence, immunohistochemistry and Western blot techniques to explore and confirme the response of nNOS-positive neurons to DA depletion. Statistical analyses were performed using SPSS 20.0 software. Results Immunofluorescence observed that There was great high co-existence rate between nNOS neurons and NPY neurons, while low co-existence rate between nNOS neurons and ChAT neurons; however, nNOS neurons with Cr or Parv neurons have no coexistence. Immunohistochemistry revealed that, nNOS neurons perform spindle and round, and in the terms of soma size （control group： 13.4±1.3 ; 6OHDA group： 13.4±1.8） , primary dendritic number of nNOS （control group ： 2.00 ± 0.94 ; 6OHDA group ： 1.77 ± 0.73 ） , control group and 6OHDA group showed no significantly difference （P≥0.05） ;as to number of nNOS, 6OHDA group （1.48 ±0.19） was significantly higher than control group （0.77±0.13）. In addition, there was a significant increment in nNOS protein expression for 6OHDA group （0.98±0.07） , compared with control group （0.56±0.04）. Conclusion It＇s evident that 6OHDA-induced DA depletion induced the characteristic response of nNOS neurons and its high protein expression, which suggested nNOS neurons may be involved in the pathological process of DA depletion-induced striatal lesion.