Drosophila Laminin: Characterization and Localization

• Published in: The Journal of cell biology Vol. 105; no. 5; pp. 2383 - 2391
• Main Authors: Fessler, L. I., Campbell, A. G., Duncan, K. G., Fessler, J. H.
• Format: Journal Article
• Language: English
• Published: New York, NY Rockefeller University Press 01.11.1987; The Rockefeller University Press
• Subjects: Amino Acids - analysis; Animals; Antibodies; Antigen-Antibody Complex; Basement membrane; Biological and medical sciences; Cell coat. Cell surface; Cell culture techniques; Cell structures and functions; Cells; Chromatography, Affinity; Chromatography, Gel; Chromatography, Ion Exchange; Collagens; Drosophila; Drosophila melanogaster; Drosophila melanogaster - analysis; Drosophilidae; Embryos; Fundamental and applied biological sciences. Psychology; Gels; Immunoglobulin G; Laminin - analysis; Laminin - isolation & purification; Microscopy, Electron; Molecular and cellular biology; Molecular Weight; Molecules; Vertebrates; Cell culture; Purification; Insecta; Immunochemistry; Drosophilidae; Characterization; Proteins; Laminin; Arthropoda; Isolation; Extracellular matrix; Invertebrata; Drosophila melanogaster; Diptera
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.105.5.2383

Drosophila laminin was isolated from the medium of Drosophila K c 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.