伪狂犬病毒法示踪成年大鼠排便反射神经通路的研究

摘要

目的 通过伪狂犬病毒示踪技术,探索成年大鼠排便反射相关的神经通路. 方法 选择正常成年SD大鼠(体重230 ～250 g)14只,随机分为实验组和对照组,实验组(10只)于末端直肠壁内注射伪狂犬病毒(Bartha株)10～ 20μL;对照组A(2只)经肛门向直肠腔内注射伪狂犬病毒(Bartha株)20 μL,对照组B(2只)于腹腔内注射伪狂犬病毒(Bartha株)20 μL.两组大鼠均于注射病毒96 h后处死,取出脑干及脊髓,进行免疫组化染色分析. 结果 实验组大鼠的脊髓各节段中均出现伪狂犬病毒阳性的神经元细胞,其中T13 ～ L1、L6 ～S1节段中病毒阳性的神经元数量较多(T13:47.5 ±2.9;L1:64.5 ±2.5;L6:30±2.1;S1:68.5±3.5),脑干Barrington氏神经核团处亦出现伪狂犬病毒阳性的神经元细胞(143.5±8.1).对照组大鼠的脊髓及脑干内均未检测出伪狂犬病毒阳性细胞. 结论 实验组的伪狂犬病毒示踪结果具有较高的特异性;成年大鼠的排便反射神经通路在脊髓内与L6至S1以及T13至L1节段关系密切,在脑干内则与Barrington氏神经核团关系密切.%Objective To explore the defecation neural pathway in spine and brainstem by inoculating pseudorabies virus (PRV,Bartha strain) into rectum in adult rats.Methods A total of 14 normal adult SD rats (male or female,weight:230 ～ 250 g) were randomly divided into two groups.The experimental group (n =10) received an inoculation of 10 ～20 μL PRV into rectal wall.The control group A had an injection of 20 μl PRV into peritoneal cavity (n =2) while the control group B 20 μL PRV into rectal cavity (n =2).At 96 h post-injection,the rats were sacrificed for harvesting spine and brainstem for immunohistochemical staining and analysis.Results PRV-immunoreactive (IR) cells were localized into almost every spinal cord in experimental rats,especially spinal segments of T13 ～ L1 and L6 ～ S1 (T13:47.5 ± 2.9;L1:64.5 ± 2.5;L6:30 ±2.1;S1:68.5 ±3.5).And PRV-IR cells were also detected in Barrington's nucleus (143.5 ± 8.1) in brainstem of experimental rats.And PRV-IR cells were not detected in control rats.Conclusions PRV transneuronal tracing has a high specificity in experimental rats.And the defecation neural pathway is closely associated with spinal segments of T13-L1 and L6-S1 and Barrington's nucleus in adult rats.