Drosophila laminin: characterization and localization.
Drosophila laminin: characterization and localization.
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DOI：
10.1083/jcb.105.5.2383
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发表时间：
1987-11
期刊：
影响因子：
--
通讯作者：
Fessler JH
中科院分区：
文献类型：
--
作者：
Fessler LI;Campbell AG;Duncan KG;Fessler JH
从果蝇Kc细胞培养培养基中分离到果蝇层粘连蛋白。通过快速沉淀、凝胶过滤和层析纯化。果蝇层粘连蛋白是一种由三条链组成的二硫分子，表观分子质量分别为400,215和185kd。在电子显微镜下，它具有与脊椎动物层粘连蛋白相似的球形结构域特征的十字形外观。给出了果蝇层粘连蛋白的氨基酸组成和凝集素结合特性。制备了果蝇层粘连蛋白的多克隆抗体，并建立了抗体的特异性。在发育的胚胎中，免疫荧光染色在发育的6 ~ 8小时进行检测；在8-9小时和更老的胚胎切片中，在内脏器官周围、肌肉周围、皮下上皮下面和神经系统的基底膜处可见免疫染色。幼虫和成虫均可见基底膜染色。在细胞胚期分离的果蝇胚胎细胞在培养中生长，免疫荧光显示，一些特异性分化细胞在培养数小时后合成层粘连蛋白。讨论了该基膜组分结构进化保存的意义。
Drosophila laminin was isolated from the medium of Drosophila Kc cell cultures. It was purified by velocity sedimentation, gel filtration, and chromatography. Drosophila laminin is a disulfide-linked molecule consisting of three chains with apparent molecular masses of 400, 215, and 185 kD. In electron micrographs, it has the cross-shaped appearance with globular domains characteristic of vertebrate laminin with closely similar dimensions. The amino acid composition and lectin-binding properties of Drosophila laminin are given. Polyclonal antibodies to Drosophila laminin were prepared and their specificity was established. In developing embryos immunofluorescence staining was detected between 6 and 8 h of development; and in sections of 8-9-h and older embryos immunostaining was seen at sites where basement membranes are present surrounding internal organs, muscles, underlying the hypodermal epithelium, and in the nervous system. Basement membrane staining was also seen in larva and adults. Cells from Drosophila embryos dissociated at the cellular blastoderm stage were grown in culture and some specific, differentiated cells synthesized laminin after several hours of culture as shown by immunofluorescence. The significance of the evolutionary conservation of the structure of this basement membrane component is discussed.