Tsui H C, Pope W B, Kim C S, Klein W L
Department of Neurobiology and Physiology, Northwestern University Institute for Neuroscience, Evanston, Illinois 60208.
J Neurobiol. 1992 Aug;23(6):720-38. doi: 10.1002/neu.480230610.
Neuritogenesis and synapse formation are transient phenomena mediated in part by filopodial attachments (Tsui, Lankford, and Klein, Proc. Natl. Acad. Sci. 82:8256-8260 1985). The attachments can be labeled by antisera against adherons, adhesive microparticles isolated from cell culture media (Tsui, Schubert, and Klein, J. Cell Biol. 106:2095-2108 1988). Here, two monoclonal antibodies raised against adherons have been found to recognize transiently expressed membrane antigens of developing avian retina. Early in development, monoclonal antibody (mAb) AD1 stained antigens that spanned the entire tissue. With time, immunoreactivity became restricted to optic fiber, ganglion cell, and inner plexiform layers. Immunoblots of embryonic day (E) 13 retina showed a broad band at 66-72 kD for particulate fractions and a fine band at 70 kD for soluble fractions. The particulate forms disappeared as retinas matured, but the soluble form did not. mAb AD2 initially labeled retina antigens of optic fiber, ganglion cell, and inner plexiform layers (IPL). Labeling in the plexiform layer showed discrete lamina. Immunoreactivity first appeared at E9, peaked at E15, and then disappeared shortly after hatching. In isolated cells, AD2 labeled small cell surface aggregates. Cytoarchitectural studies, using whole-mount transmission electron microscopy, showed AD2 antigen in cell surface microfilaments, including some that joined filopodia together. The adheron antigens recognized by mAbs AD1 and AD2 thus were (1) topographically restricted; (2) associated with cell surfaces; and (3) developmentally down-regulated. This pattern suggests a role in developmentally transient cell surface phenomena, such as neurite extension or junction biogenesis.
神经发生和突触形成是部分由丝状伪足附着介导的短暂现象(Tsui、Lankford和Klein,《美国国家科学院院刊》82:8256 - 8260,1985年)。这些附着可以用针对粘附子的抗血清进行标记,粘附子是从细胞培养基中分离出的粘性微粒(Tsui、Schubert和Klein,《细胞生物学杂志》106:2095 - 2108,1988年)。在此,已发现两种针对粘附子产生的单克隆抗体可识别发育中的鸡视网膜瞬时表达的膜抗原。在发育早期,单克隆抗体(mAb)AD1染色的抗原遍布整个组织。随着时间推移,免疫反应性局限于视神经纤维层、神经节细胞层和内网状层。胚胎第13天视网膜的免疫印迹显示,颗粒部分在66 - 72 kD处有一条宽带,可溶部分在70 kD处有一条细带。随着视网膜成熟,颗粒形式消失,但可溶形式未消失。mAb AD2最初标记视神经纤维层、神经节细胞层和内网状层(IPL)的视网膜抗原。在网状层的标记显示出离散的薄片。免疫反应性在胚胎第9天首次出现,在胚胎第15天达到峰值,然后在孵化后不久消失。在分离的细胞中,AD2标记小的细胞表面聚集体。使用整装透射电子显微镜进行的细胞结构研究显示,AD2抗原存在于细胞表面微丝中,包括一些将丝状伪足连接在一起的微丝。因此,mAbs AD1和AD2识别的粘附子抗原具有以下特点:(1)在拓扑学上受到限制;(2)与细胞表面相关;(3)在发育过程中下调。这种模式表明其在发育短暂的细胞表面现象中起作用,如神经突延伸或连接形成。