Gremo F, Kobiler D, Barondes S H
J Cell Biol. 1978 Nov;79(2 Pt 1):491-9. doi: 10.1083/jcb.79.2.491.
We determined the cellular localization of an endogenous lectin at various times during the development of a well-characterized region of chick brain, the optic tectum. This lectin is a carbohydrate-binding protein that interacts with lactose and other saccharides, undergoes striking changes in specific activity with development, and has previously been purified by affinity chromatography from extracts of embryonic chick brain and muscle. Cellular localization in the tectum was done by indirect immunofluoresecent staining, using immunoglobulin G derived from an antiserum raised against pure lectin. No lectin was detectable in the optic tectum examined at 5 days of embryonic development. From approximately 7 days of development, neuronal cell bodies and fibers were labeled by the antibody; and extracts of tectum contained hemagglutination activity that could be inhibited by lactose or by the antiserum. Lectin remained present in many tectal neuronal layers after hatching; but in 2-month-old chicks it was sparse or absent in most of the tectum except for prominent labeling of fibers in the stratum album centrale. The initial appearance of lectin in the optic tectum was not dependent on innervation by optic nerve fibers since bilateral enucleation during embryogenesis did not affect it. Lectin was detectable on the surface of embryonic optic tectal neurons dissociated with a buffer containing EDTA.
我们确定了一种内源性凝集素在鸡脑一个特征明确的区域——视顶盖发育过程中不同时间点的细胞定位。这种凝集素是一种与乳糖及其他糖类相互作用的碳水化合物结合蛋白,其比活性在发育过程中发生显著变化,并且此前已通过亲和层析从胚胎鸡脑和肌肉提取物中纯化出来。通过间接免疫荧光染色进行视顶盖中的细胞定位,所用的免疫球蛋白G源自针对纯凝集素产生的抗血清。在胚胎发育第5天检查的视顶盖中未检测到凝集素。从大约发育第7天开始,神经元细胞体和纤维被抗体标记;并且视顶盖提取物含有可被乳糖或抗血清抑制的血凝活性。孵化后,凝集素仍存在于许多视顶盖神经元层中;但在2月龄雏鸡中,除了中央白质层中纤维有明显标记外,视顶盖的大部分区域凝集素稀少或不存在。视顶盖中凝集素的最初出现不依赖于视神经纤维的支配,因为胚胎期双侧眼球摘除并不影响它。在含有EDTA的缓冲液中解离的胚胎视顶盖神经元表面可检测到凝集素。