Summary
By using an in vitro functional assay, we have shown that Drosophila embryonic cells possess Ca2+-dependent adhesive sites, which resemble in many respects those described for vertebrate cells and tissues. The cells, obtained by mechanical disruption of gastrulastage embryos, form aggregates within 30 min when maintained under constant rolling. The aggregation is completely dependent on the presence of Ca2+ in the medium. In its absence, the cells remain dispersed but the process is reversible by readdition of Ca2+. In addition the aggregation is temperature-dependent. No aggregation occurs at 4° C but it can be restored by raising the temperature to 25° C. These properties are characteristic of these cells: established cell lines do not aggregate under the same conditions and mixing of cell lines and embryonic cells does not result in chimeric aggregates, thus pointing towards cell-type selectivity with respect to aggregability. Observations in electron microscopy have shown that the embryonic cells in the aggregates tightly adhere to one another and form, as early as after 30 min, maculae adherens junctions. Drosophila embryonic cells have adhesion sites that are protected from trypsin proteolysis in the presence of Ca2+ and sensitive in its absence. The cells' aggregation can be inhibited by a mouse antiserum directed against cell-surface components and a good correlation exists between neutralization of the inhibitory activity of the antiserum and the presence of trypsin-sensitive sites on the cells. These data are in favour of cell-cell adhesion mediated by specific adhesion proteins.
Similar content being viewed by others
References
Anderson H (1988) Drosophila adhesion molecules and neural development. Trends Neurosci 11:472–475
Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–252
Burt R, Gierer A (1979) Age specific cell sorting within aggregates of chick neural retina cells. Roux's Arch Dev Biol 187:367–373
Choi YS, Gumbiner B (1989) Expression of cell adhesion molecule E-cadherin in Xenopus embryos begins at gastrulation and predominates in the ectoderm. J Cell Biol 108:2449–2458
Cunningham BA, Hemperly JJ, Murray BA, Prediger EA, Brackenbury R, Edelman GM (1987) Neural cell adhesion molecule: structure, immunoglobulin-like domains, cell surface modulation, and alternative RNA splicing. Science 236:799–806
Currie DA, Milner MJ, Evans CW (1988) The growth and differentiation in vitro of leg and wing imaginal disc cells from Drosophila melanogaster. Development 102:805–814
Edelman GM (1986) Cell adhesion molecules in the regulation of animal form and tissue pattern. Annu Rev Cell Biol 2:81–117
Edelman GM, Murray BA, Mege R, Cunningham BA, Gallin WJ (1987) Cellular expression of liver and neural cell adhesion molecules after transfection with their cDNAs results in specific cell-cell binding. Proc Natl Acad Sci USA 84:8502–8506
Fausto-Sterling A, Hsieh L (1987) In vitro culture of Drosophila imaginal disc cells: aggregation, sorting out, and differentiation abilities. Dev Biol 120:284–293
Fausto-Sterling A, Muckenthaler FA, Hsieh L, Rosenblatt PL (1985) Some determinants of cellular adhesiveness in an embryonic cell line from Drosophila melanogaster. J Exp Zool 234:47–55
Fehon RG, Schubiger G (1985) Dissociation and sorting out of Drosophila imaginal disc cells. Dev Biol 108:465–473
Furst A, Mahowald AP (1985) Differentiation of primary embryonic neuroblasts in purified neural cell cultures from Drosophila. Dev Biol 109:184–192
Galewsky S, Rickoll WL (1989) 20-hydroxyecdysone induced aggregation of Drosophila S3 cells is inhibited by antibodies to a hormone-dependent extracellular glycoprotein. Roux's Arch Dev Biol 198:14–18
Gauger A, Fehon RG, Schubiger G (1985) Preferential binding of imaginal disc cells to embryonic segments of Drosophila. Nature 313:395–397
Gratecos D, Astier M, Sémériva M (1987) A new appproach to monoclonal antibody production. In vitro immunization with antigens on nitrocellulose using Drosophila myosin heavy chain as an example. J Immunol Methods 103:169–178
Gratecos D, Naidet C, Astier M, Thiery JP, Sémériva M (1988) Drosophila fibronectin: a protein that shares properties similar to those of its mammalian homologue. EMBO J 7:215–223
Harrelson AL, Goodman CS (1988) Growth cone guidance in insects: fasciclin II is a member of the immunoglobulin superfamily. Science 242:700–708
Hatta K, Nose A, Nagafuchi A, Takeichi M (1988) Cloning and expression of cDNA encoding a neural calcium-dependent cell adhesion molecule: its identity in the cadherin gene family. J Cell Biol 106:873–881
Hay ED (1968) Organization and fine structure of epithelium and mesenchyme in the developing chick embryo. In: Fleishmajer R, Billingham R (eds) ‘Epithelial-mesenchymal interactions’. Williams and Wilkins, Baltimore, Maryland, pp 31–55
Hubbard AL, Cohn ZA (1972) The enzymatic iodination of the cell membrane. J Cell Biol 55:390–405
Klämbt C, Müller S, Lützelschwab R, Rossa R, Totzke F, Schmidt O (1989) The Drosophila melanogaster l(2)gl gene encodes a protein homologous to the cadherin cell adhesion molecule family. Dev Biol 133:425–436
Laemmli UK (1970) Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227:680–685
Moscona A, Moscona H (1952) Dissociation and aggregation of cells from organ rudiments of the early chick embryos. J Anat 86:287–301
Nagafuchi A, Shirayoshi Y, Okazaki K, Yasuda K, Takeichi M (1987) Transformation of cell adhesion properties by exogenously introduced E-cadherin cDNA. Nature 329:341–343
Nose A, Nagafuchi A, Takeichi M (1988) Expressed recombinant cadherins mediate cell sorting in model systems. Cell 54:993–1001
Ozawa M, Baribault H, Kemler R (1989) The cytoplasmic domain of the cell adhesion molecule uvomorulin associates with three independent proteins structurally related in different species. EMBO J 8:1711–1717
Rickoll WL, Counce SJ (1980) Morphogenesis in the embryo of Drosophila melanogaster. Germ band extension. Roux's Arch Dev Biol 188:163–177
Sang JH (1981) Drosophila cells and cell lines. Adv Cell Cult 1:125–182
Seecof RL (1979) Preparation of cell cultures from Drosophila melanogaster embryos. TCA Man 5:1019–1022
Seecof RL, Alleaume N, Teplitz RL, Gerson I (1971) Differentiation of neurons and myocytes in cell cultures made from Drosophila gastrulae. Exp Cell Res 69:161–173
Seecof RL, Donady JJ, Teplitz RL (1973) Differentiation of Drosophila neuroblasts to form ganglion-like clusters of neurons in vitro. Cell Differ 2:143–149
Seeger MA, Haffley L, Kaufman TC (1988) Characterization of amalgam: a member of the immunoglobulin superfamily of Drosophila. Cell 55:589–600
Sémériva M, Naidet C, Krejci E, Gratecos D (1989) Towards the molecular biology of cell adhesion in Drosophila. Trends Genet 5:24–28
Shields G, Sang JH (1977) An improved medium for the cultivation of Drosophila cells. Drosophila Inf Ser 52:161
Takeichi M (1977) Functional correlation between cell adhesive properties and some cell surface proteins. J Cell Biol 75:464–474
Takeichi M (1988) The cadherins: cell-cell adhesion molecules controlling animal morphogenesis. Development 102:639–655
Takeichi M, Hatta K, Nagafuchi A (1985) Selective cell-adhesion mechanism: role of the calcium-dependent cell adhesion system. In: Edelman GM (ed) Molecular determinants of animal form. AR Liss, New-York, pp 223–233
Townes PL, Holtfreter J (1955) Directed movements and selective adhesion of embryonic amphibian cells. J Exp Zool 128:53–120
Trinkaus JP (1984) Cells into organs, Prentice Hall, Englewood Cliffs, NJ
Ui K, Ueda R, Miyake T (1988) In vitro cultures of cells from different kinds of imaginal discs of Drosophila melanogaster. Jpn J Genet 63:33–41
Wu CF, Suzuki N, Poo MM (1983) Dissociated neurons from normal and mutant Drosophila larval central nervous system in cell culture. J Neurosci 3:1888–1899
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Gratecos, D., Krejci, E. & Sémériva, M. Calcium-dependent adhesion of Drosophila embryonic cells. Roux's Arch Dev Biol 198, 411–419 (1990). https://doi.org/10.1007/BF00376160
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00376160