Aguado F, Majó G, Ruiz-Montasell B, Canals J M, Casanova A, Marsal J, Blasi J
Department de Biologia Cel.lular i Anatomia Patològica, Facultat de Medicina, Universitat de Barcelona, Hospital de Bellvitge, Spain.
Eur J Cell Biol. 1996 Apr;69(4):351-9.
A growing body of evidence indicates that the fundamental molecular mechanism of exocytosis in the secretory pathway may be structurally similar in all eukaryotic cells. The synaptosomal-associated protein of 25 kDa (SNAP-25) is a plasma membrane protein involved in regulated exocytosis in neurons. In order to compare exocytotic components in neurons and endocrine cells, we have analyzed the expression of SNAP-25 in the rat anterior pituitary. Western blotting analysis documented the presence of SNAP-25 in anterior pituitary homogenates and cultured anterior pituitary cells. In addition to SNAP-25, other neuronal proteins involved in exocytosis (syntaxin, VAMP/synaptobrevin and Rab3A) were also detected in the anterior pituitary. The specific expression of SNAP-25 mRNA in anterior pituitary cells was also corroborated by Northern analysis. SNAP-25 immunoreactivity was located at the plasma membrane of endocrine anterior pituitary cells. Characteristically, patches of fine punctate deposits exhibited intense SNAP-25 immunoreactivity. Double-labeling immunocytochemistry revealed that SNAP-25 was mainly associated with gonadotroph cell populations. Furthermore, we demonstrate that in the anterior pituitary, SNAP-25 is selectively cleaved by clostridial neurotoxins. In conclusion, our results establish the presence of SNAP-25 in secretory anterior pituitary cells and suggest a potential role of this protein in the secretion of adenohypophysial hormone.
越来越多的证据表明,分泌途径中胞吐作用的基本分子机制在所有真核细胞中可能在结构上相似。25 kDa的突触体相关蛋白(SNAP-25)是一种参与神经元中调节性胞吐作用的质膜蛋白。为了比较神经元和内分泌细胞中的胞吐成分,我们分析了大鼠垂体前叶中SNAP-25的表达。蛋白质印迹分析证明垂体前叶匀浆和培养的垂体前叶细胞中存在SNAP-25。除了SNAP-25,在垂体前叶中还检测到了其他参与胞吐作用的神经元蛋白( syntaxin、VAMP/突触小泡蛋白和Rab3A)。Northern分析也证实了垂体前叶细胞中SNAP-25 mRNA的特异性表达。SNAP-25免疫反应性位于垂体前叶内分泌细胞的质膜上。特征性地,细小点状沉积物斑块表现出强烈的SNAP-25免疫反应性。双标记免疫细胞化学显示SNAP-25主要与促性腺激素细胞群体相关。此外,我们证明在垂体前叶中,SNAP-25被梭菌神经毒素选择性切割。总之,我们的结果证实了分泌性垂体前叶细胞中存在SNAP-25,并表明该蛋白在腺垂体激素分泌中具有潜在作用。