Centre for Integrative Physiology, University of Edinburgh, Edinburgh, UK.
J Neuroendocrinol. 2012 Apr;24(4):629-41. doi: 10.1111/j.1365-2826.2011.02237.x.
In magnocellular neurones of the supraoptic nucleus (SON), the neuropeptides vasopressin and oxytocin are synthesised and packaged into large dense-cored vesicles (LDCVs). These vesicles undergo regulated exocytosis from nerve terminals in the posterior pituitary gland and from somata/dendrites in the SON. Regulated exocytosis of LDCVs is considered to involve the soluble N-ethylmaleimide sensitive fusion protein attachment protein receptor (SNARE) complex [comprising vesicle associated membrane protein 2 (VAMP-2), syntaxin-1 and soluble N-ethylmaleimide attachment protein-25 (SNAP-25)] and regulatory proteins [such as synaptotagmin-1, munc-18 and Ca(2+) -dependent activator protein for secretion (CAPS-1)]. Using fluorescent immunocytochemistry and confocal microscopy, in both oxytocin and vasopressin neurones, we observed VAMP-2, SNAP-25 and syntaxin-1-immunoreactivity in axon terminals. The somata and dendrites contained syntaxin-1 and other regulatory exocytosis proteins, including munc-18 and CAPS-1. However, the distribution of VAMP-2 and synaptotagmin-1 in the SON was limited to putative pre-synaptic contacts because they co-localised with synaptophysin (synaptic vesicle marker) and had no co-localisation with either oxytocin or vasopressin. SNAP-25 immunoreactivity in the SON was limited to glial cell processes and was not detected in oxytocin or vasopressin somata/dendrites. The present results indicate differences in the expression and localisation of exocytosis proteins between the axon terminals and somata/dendritic compartment. The absence of VAMP-2 and SNAP-25 immunoreactivity from the somata/dendrites suggests that there might be different SNARE protein isoforms expressed in these compartments. Alternatively, exocytosis of LDCVs from somata/dendrites may use a different mechanism from that described by the SNARE complex theory.
在视上核(SON)的大细胞神经元中,神经肽血管加压素和催产素被合成并包装到大型致密核心囊泡(LDCV)中。这些囊泡从垂体后叶的神经末梢以及 SON 中的体/树突中通过受调节的胞吐作用释放。LDCV 的受调节胞吐作用被认为涉及可溶性 N-乙基马来酰亚胺敏感融合蛋白附着蛋白受体(SNARE)复合物[包括囊泡相关膜蛋白 2(VAMP-2)、突触融合蛋白 1(syntaxin-1)和可溶性 N-乙基马来酰亚胺附着蛋白 25(SNAP-25)]和调节蛋白[如突触结合蛋白 1、Munc-18 和钙依赖性分泌激活蛋白(CAPS-1)]。使用荧光免疫细胞化学和共聚焦显微镜,在催产素和血管加压素神经元中,我们观察到 VAMP-2、SNAP-25 和 syntaxin-1-免疫反应性在轴突末端。体和树突包含 syntaxin-1 和其他调节胞吐作用的蛋白质,包括 Munc-18 和 CAPS-1。然而,VAMP-2 和突触结合蛋白 1 在 SON 中的分布仅限于假定的突触前接触,因为它们与突触小泡蛋白(突触小泡标记物)共定位,并且与催产素或血管加压素没有共定位。SNAP-25 在 SON 中的免疫反应性仅限于神经胶质细胞过程,在催产素或血管加压素体/树突中未检测到。目前的结果表明,轴突末端和体/树突区室之间的胞吐作用蛋白的表达和定位存在差异。VAMP-2 和 SNAP-25 免疫反应性在体/树突中缺失表明,这些区室中可能表达不同的 SNARE 蛋白同工型。或者,LDCV 从体/树突的胞吐作用可能使用不同于 SNARE 复合物理论所描述的机制。