Feldman E L, Axelrod D, Schwartz M, Heacock A M, Agranoff B W
J Neurobiol. 1981 Nov;12(6):591-8. doi: 10.1002/neu.480120607.
Explant culture of the adult goldfish retina results in vigorous neuritic growth, provided that the optic nerve has been previously crushed in vivo. The site of new membrane addition in these growing neurites was studied with a a membrane-binding lectin Concanavalin A (Con A) in conjunction with a fluorescent indirect antibody method. Explant cultures were labeled with Con A followed by rabbit antibodies to Con A. This treatment was shown to immobilize Con A receptors while permitting growth. Twenty-four hours later, the washed preparation was treated with fluorescent goat antirabbit antibodies. The bound fluorescent antibodies appeared as a pattern of contiguous fluorescent speckles confined to the area of old growth while regions near the distal end of the neurite were not fluorescent. These results are compatible with the hypothesis that new membrane is added in the region of the growing tip of the neurite.
成年金鱼视网膜的外植体培养可导致旺盛的神经突生长,前提是视神经此前已在体内被切断。利用膜结合凝集素伴刀豆球蛋白A(Con A)结合荧光间接抗体法,研究了这些生长中的神经突上新膜添加的部位。外植体培养物先用Con A标记,然后用抗Con A的兔抗体处理。结果表明,这种处理可使Con A受体固定,同时允许生长。24小时后,洗涤后的标本用荧光山羊抗兔抗体处理。结合的荧光抗体呈现为连续荧光斑点的模式,局限于旧生长区域,而神经突远端附近的区域没有荧光。这些结果与神经突生长尖端区域添加新膜的假说相符。