Department of Neuroscience, University of Pittsburgh, Pittsburgh, PA 15260, United States.
Neuroscience. 2012 Dec 27;227:247-59. doi: 10.1016/j.neuroscience.2012.09.049. Epub 2012 Oct 3.
C1 catecholamine neurons reside within the rostroventrolateral medulla (RVLM), an area that plays an integral role in blood pressure regulation through reticulospinal projections to sympathetic preganglionic neurons in the thoracic spinal cord. In a previous investigation we mapped the efferent projections of C1 neurons, documenting supraspinal projections to cell groups in the preautonomic network that contribute to the control of cardiovascular function. Light microscopic study also revealed putative local circuit connections within RVLM. In this investigation we tested the hypothesis that RVLM C1 neurons elaborate a local circuit synaptic network that permits communication between C1 neurons giving rise to supraspinal and reticulospinal projections. A replication defective lentivirus vector that expresses enhanced green fluorescent protein (EGFP) under the control of a synthetic dopamine beta hydroxylase (DβH) promoter was used to label C1 neurons and their processes. Confocal fluorescence microscopy demonstrated thin varicose axons immunopositive for EGFP and tyrosine hydroxylase that formed close appositions to C1 somata and dendrites throughout the rostrocaudal extent of the C1 area. Dual-labeled electron microscopic analysis revealed axosomatic, axodendritic and axospinous synaptic contacts with C1 and non-C1 neurons with a distribution recapitulating that observed in the light microscopic analysis. Labeled boutons were large, contained light axoplasm, lucent spherical vesicles, and formed asymmetric synaptic contacts. Collectively these data demonstrate that C1 neurons form a synaptic network within the C1 area that may function to coordinate activity among projection-specific subpopulations of neurons. The data also suggest that the boundaries of RVLM should be defined on the basis of function criteria rather than the C1 phenotype of neurons.
C1 儿茶酚胺神经元位于延髓腹外侧区(RVLM),该区域通过网状脊髓投射到胸段脊髓交感节前神经元,在血压调节中起着重要作用。在之前的一项研究中,我们绘制了 C1 神经元的传出投射图,记录了向自主前网络中细胞群的中枢投射,这些细胞群有助于心血管功能的控制。光镜研究还揭示了 RVLM 内的假定局部回路连接。在这项研究中,我们检验了这样一个假设,即 RVLM C1 神经元形成一个局部回路突触网络,允许起源于中枢和网状脊髓投射的 C1 神经元之间进行通讯。复制缺陷型慢病毒载体,在合成多巴胺β羟化酶(DβH)启动子的控制下表达增强型绿色荧光蛋白(EGFP),用于标记 C1 神经元及其过程。共聚焦荧光显微镜显示,免疫阳性的 EGFP 和酪氨酸羟化酶的细而曲折的轴突形成与 C1 体和树突的紧密贴合,贯穿 C1 区的头尾延伸。双重标记的电子显微镜分析显示,与 C1 和非 C1 神经元形成轴体、轴树突和轴棘突突触接触,其分布与光镜分析观察到的分布相似。标记的终末是大的,含有轻轴浆、透明的球形小泡,并形成不对称的突触接触。这些数据共同表明,C1 神经元在 C1 区内形成一个突触网络,可能用于协调特定神经元亚群的活动。这些数据还表明,RVLM 的边界应该基于功能标准而不是神经元的 C1 表型来定义。