Jena Bhanu P, Cho Sang-Joon, Jeremic Aleksandar, Stromer Marvin H, Abu-Hamdah Rania
Department of Physiology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.
Biophys J. 2003 Feb;84(2 Pt 1):1337-43. doi: 10.1016/S0006-3495(03)74949-2.
Earlier studies using atomic force microscopy (AFM) demonstrated the presence of fusion pores at the cell plasma membrane in a number of live secretory cells, revealing their morphology and dynamics at nm resolution and in real time. Fusion pores were stable structures at the cell plasma membrane where secretory vesicles dock and fuse to release vesicular contents. In the present study, transmission electron microscopy confirms the presence of fusion pores and reveals their detailed structure and association with membrane-bound secretory vesicles in pancreatic acinar cells. Immunochemical studies demonstrated that t-SNAREs, NSF, actin, vimentin, alpha-fodrin and the calcium channels alpha1c and beta3 are associated with the fusion complex. The localization and possible arrangement of SNAREs at the fusion pore are further demonstrated from combined AFM, immunoAFM, and electrophysiological measurements. These studies reveal the fusion pore or porosome to be a cup-shaped lipoprotein structure, the base of which has t-SNAREs and allows for docking and release of secretory products from membrane-bound vesicles.
早期使用原子力显微镜(AFM)的研究表明,在许多活的分泌细胞的细胞质膜上存在融合孔,揭示了它们在纳米分辨率下的形态和动力学,并能实时观察。融合孔是细胞质膜上的稳定结构,分泌小泡在此停靠并融合以释放小泡内容物。在本研究中,透射电子显微镜证实了融合孔的存在,并揭示了它们在胰腺腺泡细胞中的详细结构以及与膜结合分泌小泡的关联。免疫化学研究表明,t-SNAREs、NSF、肌动蛋白、波形蛋白、α- fodrin以及钙通道α1c和β3与融合复合体相关。通过结合AFM、免疫AFM和电生理测量,进一步证明了SNAREs在融合孔处的定位和可能的排列。这些研究揭示融合孔或孔体是一种杯状脂蛋白结构,其底部含有t-SNAREs,允许膜结合小泡停靠并释放分泌产物。