大鼠脊髓及脑干内膀胱支配中枢的跨突触示踪研究

Transsynaptic tracing of CNS neural circuitry involved in the innervation of bladder function in the adult rat brainstem and spinal cord

目的跨突触示踪正常大鼠脊髓和脑干内膀胱支配相关中枢,为进一步阐明膀胱功能重建术后中枢重塑奠定研究基础。方法成年雌性SD大鼠15只,膀胱壁内分三个点共注射GFP-PRV 4.5μ（l1×108PFU/ml）。注射后不同时间（72、84、96、108、120 h）分别取大脑、脊髓及背根神经节,荧光显微镜下观察标记结果。结果病毒注射后72～120 h,阳性神经元主要分布于L6～S1、L1～L2脊神经背根神经节;L6～S1、L1～L2脊髓节段骶副交感核、中间外侧核及后连合核;脑干的巴氏核、中缝巨细胞核、巨细胞网状核、锥旁网状结构、去甲肾上腺素能细胞群A5及A7、蓝斑、中脑导水管周围灰质和红核腹侧区域。结论本实验中标记的相关核团与膀胱存在解剖上的神经联系,可能直接或间接参与膀胱的支配。

Objective To identify the CNS neural circuitry involved in the innervation of bladder function in the adult rat brainstem and spinal cord using pseudorabies virus, a transsynaptic tracer, so as to lay down the basis for further studying of brain functional reorganization after bladder function reconstruction. Methods GFP-PRV 4.5ul （ 1 × 108 PFU/ml）was injected into the bladder wall of 15 adult female SD rats at 3 different sites. The distribution of virus-infected neurons in the brainstem, spinal cord and dorsal root ganglion were observed under fluorescent microscope at various intervals （72h, 84h, 96h, 108h, 120h） following the PRV injection. Results Fluorescence positive neurons were mainly present in L6～S1, LI～L2 dorsal root ganglion; sacral parasympathetic nucleus, intermediolateral cell column and dorsal commissural in L6～S1 and L1～L2 spinal cord segments; Barrington＇s nucleus, nucleus raphe magnus, the gigantocelluar reticular nucleus, the parapyramidal reticular formation, noradrenergic cell groups A5 and A7, locus coeruleus, periaqueductal gray and the ventral region of red nucleus in brainstem. Conoltusion The structures labeled in spinal cord and brainstem are synaptically connected with the bladder and presumably involved in the neural control of the bladder.