Fey J, Hausen P
Max-Planck-Institut für Entwicklungsbiologie, Abt. Zellbiologie, Tübingen, Federal Republic of Germany.
Differentiation. 1990 Feb;42(3):144-52. doi: 10.1111/j.1432-0436.1990.tb00755.x.
A cloned cDNA fragment, homologous to domain II of the mouse laminin B1 chain, was obtained from a Xenopus neurula cDNA library. Using this probe Northern-blot analysis over the course of embryogenesis revealed a first signal of a laminin transcript at midgastrula (stage 11). Affinity-purified polyclonal antibodies directed against Xenopus laminin were obtained via a lacZ fusion protein. Immunohistology demonstrated the appearance of the antigen at about one developmental stage after the detection of the transcript. At stage 12 1/2 a faint immunofluorescence staining surrounded the developing notochord and somitogenic mesodermal cells. Thereafter laminin appeared in distinct locations outlining notochord, somites and neural tube. The epidermal basement membrane seemed to be endowed with laminin only at the relatively late postneurula stage. The observation that laminin could not be detected before stage 12 1/2 is discussed with respect to a proposed role of laminin in the gastrulation process.
从小爪蟾神经胚cDNA文库中获得了一个与小鼠层粘连蛋白B1链结构域II同源的克隆cDNA片段。使用该探针在胚胎发育过程中进行Northern杂交分析，结果显示在原肠胚中期（11期）首次出现层粘连蛋白转录本信号。通过lacZ融合蛋白获得了针对小爪蟾层粘连蛋白的亲和纯化多克隆抗体。免疫组织学显示，在检测到转录本后约一个发育阶段出现该抗原。在12.5期，微弱的免疫荧光染色围绕着正在发育的脊索和体节生成中胚层细胞。此后，层粘连蛋白出现在勾勒出脊索、体节和神经管的不同位置。表皮基底膜似乎仅在神经胚后期才具有层粘连蛋白。关于层粘连蛋白在原肠胚形成过程中的假定作用，讨论了在12.5期之前未检测到层粘连蛋白这一观察结果。
