Cleary L J, Schwartz J H
Howard Hughes Medical Institute, Columbia University College of Physicians and Surgeons, New York, NY 10032.
J Comp Neurol. 1987 Sep 1;263(1):92-105. doi: 10.1002/cne.902630108.
In order to ascertain the form in which newly synthesized membrane is moved by fast axonal transport, we examined the distribution of label along an axon of the identified giant serotoninergic neuron (GCN) in the cerebral ganglion of Aplysia californica. Membrane glycoproteins were labeled by intrasomatic injection of 3H-fucose, and segments of GCN's axon in the posterior lip nerve containing the transported organelles were examined at 1, 5, 15, and 24 hours by quantitative electron-microscopic autoradiography. To show that membrane which is rapidly transported is contained only in discrete organelles rather than in continuous sheets of axoplasmic reticulum, we systematically varied conditions of fixation. We found that we could distinguish vesicles from axoplasmic reticulum most reliably in axons fixed with 2% formaldehyde and 2% glutaraldehyde in cacodylate buffer. At short times after intrasomatic injection of 3H-fucose, dense-cored vesicles and multivesicular tubules were the only axonal organelles labeled.
为了确定新合成的膜通过快速轴突运输移动的形式,我们检查了加州海兔脑神经节中已鉴定的巨型5-羟色胺能神经元(GCN)轴突上标记物的分布。通过向胞内注射3H-岩藻糖来标记膜糖蛋白,并在1、5、15和24小时时,通过定量电子显微镜放射自显影术检查后唇神经中含有运输细胞器的GCN轴突段。为了表明快速运输的膜仅包含在离散的细胞器中,而不是在连续的轴质网片中,我们系统地改变了固定条件。我们发现,在用2%甲醛和2%戊二醛的二甲胂酸盐缓冲液固定的轴突中,我们能够最可靠地区分囊泡和轴质网。在向胞内注射3H-岩藻糖后的短时间内,致密核心囊泡和多泡小管是唯一被标记的轴突细胞器。