利用突触小泡蛋白-GFP融合蛋白对活的秀丽隐杆线虫中的突触特化进行可视化观察。

Visualization of synaptic specializations in live C. elegans with synaptic vesicle protein-GFP fusions.

作者信息

Nonet M L

机构信息

Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, MO 63110, USA.

出版信息

J Neurosci Methods. 1999 Jul 1;89(1):33-40. doi: 10.1016/s0165-0270(99)00031-x.

DOI:10.1016/s0165-0270(99)00031-x

PMID:10476681

Abstract

Synaptic specializations are difficult to visualize at the light microscope level in living preparations. To circumvent this problem, synaptic vesicle proteins were fused to green fluorescent protein (GFP) and expressed in C. elegans neurons. C. elegans synaptobrevin-GFP and synaptogyrin-GFP fusion proteins were observed to target to synaptic sites. This localization allowed the visualization of synaptic specializations in living animals with light microscopy. Restricted expression of synaptobrevin-GFP fusions in subsets of neurons enables the visualization of individual presynaptic varicosities. The cell biology underlying the sorting of synaptic vesicle proteins, trafficking of vesicles to terminals, and the development of presynaptic specializations is now more amenable to forward genetic analysis using these synaptic markers.

摘要

在活体标本中，突触特化结构在光学显微镜水平下很难可视化。为了解决这个问题，将突触小泡蛋白与绿色荧光蛋白（GFP）融合，并在秀丽隐杆线虫神经元中表达。观察到秀丽隐杆线虫的突触融合蛋白-GFP和突触素-GFP融合蛋白靶向突触部位。这种定位使得在活体动物中通过光学显微镜观察突触特化结构成为可能。突触融合蛋白-GFP融合蛋白在神经元亚群中的限制性表达能够可视化单个突触前膨体。现在，利用这些突触标记物，对突触小泡蛋白分选、小泡向终末运输以及突触前特化结构形成的细胞生物学基础进行正向遗传学分析变得更加容易。