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用单克隆抗体D3检测成年金鱼视网膜顶盖通路中N-CAM的时空分布。

The spatiotemporal distribution of N-CAM in the retinotectal pathway of adult goldfish detected by the monoclonal antibody D3.

作者信息

Bastmeyer M, Schlosshauer B, Stuermer C A

机构信息

Friedrich-Miescher-Laboratorium, Max-Planck-Gesellschaft, Tübingen, FRG.

出版信息

Development. 1990 Feb;108(2):299-311. doi: 10.1242/dev.108.2.299.

Abstract

The spatiotemporal distribution of neural cell adhesion molecule (N-CAM) in the retinotectal system of adult goldfish was assessed by immunofluorescence using the monoclonal antibody (Mab) D3 against chick N-CAM. In immunoblots with extracts of cell surface membranes of fish brains, Mab D3 recognized a prominent band at 170K and a weak band at 130K (K = 10(3) Mr). N-CAM immunofluorescence on cells was restricted to the marginal growth zones of the retina and the tectum and, in normal fish, to the youngest axons from the new ganglion cells of the peripheral retinal margin. In fish with previously transected optic nerves (ONS), Mab D3 staining was found transiently on all axons from the site of the cut into the retinorecipient layers of the tectum, but disappeared from these axons 450 days after ONS. Growing retinal axons in vitro exhibited N-CAM immunofluorescence throughout their entire extent, including their growth cones. Glial cells cultured from regenerating optic nerves were, however, unlabeled. These data are consistent with the idea that N-CAM is involved in adhesive interactions of growing axons. The temporally regulated expression of N-CAM on the new retinal axons may contribute to the creation of the age-related organization of the axons in the retinotectal pathway of fish.

摘要

利用抗鸡神经细胞黏附分子(N-CAM)的单克隆抗体(Mab)D3,通过免疫荧光法评估成年金鱼视网膜顶盖系统中N-CAM的时空分布。在用鱼脑细胞膜提取物进行的免疫印迹中,Mab D3识别出一条位于170K的显著条带和一条位于130K的弱条带(K = 10(3) Mr)。细胞上的N-CAM免疫荧光仅限于视网膜和顶盖的边缘生长区,在正常鱼中,仅限于来自周边视网膜边缘新神经节细胞的最年轻轴突。在先前切断视神经(ONS)的鱼中,在从切断部位到顶盖视网膜接受层的所有轴突上短暂发现Mab D3染色,但在ONS后450天这些轴突上的染色消失。体外生长的视网膜轴突在其整个长度包括生长锥上都表现出N-CAM免疫荧光。然而,从再生视神经培养的神经胶质细胞未被标记。这些数据与N-CAM参与生长轴突的黏附相互作用这一观点一致。N-CAM在新视网膜轴突上的时间调控表达可能有助于在鱼的视网膜顶盖通路中形成与年龄相关的轴突组织。

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