Peng Y W, Lam D M
Alice R. McPherson Laboratory of Retina Research, Baylor College of Medicine, The Woodlands, TX 77381.
Vis Neurosci. 1991 Apr;6(4):357-70. doi: 10.1017/s0952523800006581.
We have produced and characterized a monoclonal antibody, AT101, which selectively labels both viable and formaldehyde-fixed horizontal cell axon terminals, but not their somas or axons, of the goldfish (Carassius auratus) retina. The antigen recognized by AT101 appears to be a cell surface glycoprotein with a molecular weight of about 35,000 Daltons, and is present exclusively or predominantly in nervous tissues of all vertebrate species examined. We have used AT101 as a probe to analyze immunocytochemically the organization of horizontal cell axon terminals (HCATs) in the adult goldfish retina, and the emergence and maturation of these terminals during retinal development. Because of continued growth at the retinal margin in adult goldfish, there is a peripheral-to-central gradient in the age of cells, with the most mature in the center and the youngest in the periphery. In the center and near periphery of the adult retina, HCATs have a fusiform morphology and form a dense network in the middle and proximal part of the inner nuclear layer. In the far peripheral retina, the axon terminals appear round or ellipsoid. The retina closest to the retinal margin is devoid of AT101 staining, indicating that either HCATs are absent or the antigen recognized by AT101 is not present on HCATs at this stage. A similar sequence of changes in staining pattern is seen during development. Although AT101 staining can first be demonstrated in the larval retina at 1 month after hatching, it appears mostly as punctate structures. At a later stage, there are round or ellipsoid structures that resemble in morphology and location (in the inner nuclear layer) those found in the far peripheral adult retina. Double-labeling experiments with AT101 and antiserum against tubulin also indicate that AT101 labels the HCATs when they first appear during development. These data suggest that the emergence and maturation of HCAT is a late event in retinal development.
我们制备并鉴定了一种单克隆抗体AT101,它能选择性地标记金鱼(Carassius auratus)视网膜中存活的和经甲醛固定的水平细胞轴突终末,但不标记其胞体或轴突。AT101识别的抗原似乎是一种分子量约为35,000道尔顿的细胞表面糖蛋白,且仅存在于或主要存在于所有被检测脊椎动物物种的神经组织中。我们使用AT101作为探针,通过免疫细胞化学方法分析成年金鱼视网膜中水平细胞轴突终末(HCATs)的组织结构,以及这些终末在视网膜发育过程中的出现和成熟情况。由于成年金鱼视网膜边缘持续生长,细胞年龄存在从周边到中央的梯度,中央的细胞最成熟,周边的细胞最年轻。在成年视网膜的中央和近周边区域,HCATs呈梭形形态,并在内核层的中部和近端形成密集网络。在远周边视网膜中,轴突终末呈圆形或椭圆形。最靠近视网膜边缘的区域没有AT101染色,这表明在这个阶段要么没有HCATs,要么AT101识别的抗原不在HCATs上。在发育过程中也观察到类似的染色模式变化序列。虽然在孵化后1个月的幼体视网膜中就可以首次检测到AT101染色,但最初它大多表现为点状结构。在后期阶段,会出现圆形或椭圆形结构,其形态和位置(在内核层)与远周边成年视网膜中的结构相似。用AT101和抗微管蛋白抗血清进行的双重标记实验也表明,AT101在HCATs发育过程中首次出现时就对其进行标记。这些数据表明,HCATs的出现和成熟是视网膜发育后期的事件。