Summary
From the central nervous system ofDrosophila melanogaster 3rd instar larvae, eight continuous cell lines have been established (named ML-DmBG1 to 8). Using ML-DmBG2, single colony isolation was carried out and six colonial clones were obtained. All reacted to the antibody to horseradish peroxidase, which is a neuronal marker in insects. Acetylcholine, a known neurotransmitter inDrosophila, was detected in three of the colonial clones by high performance liquid chromatography. Therefore, it is concluded that the established colonial clones are neural cells originating in the larval central nervous system. Among them, some variation was observed with respect to morphology, acetylcholine content, and reactivity to anti-HRP. The variation may reflect the heterogeneity of cells composing the central nervous system.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Cross, D. P.; Sang, J. H. Cell culture of individualDrosophila embryos I. Development of wild-type cultures. J. Embryol. Exp. Biol. 45:161–172; 1978.
Currie, D. A.; Milner, M. J.; Evans, C. W. The growth and differentiationin vitro of leg and wing imaginal disc cells fromDrosophila melanogaster. Development 102:805–814; 1988.
Davis, K. T.; Shearn, A.In vitro growth of imaginal disks fromDrosophila melanogaster. Science 196:438–440; 1977.
Echalier, G.; Ohanessian, A.In vitro culture ofDrosophila melanogaster embryonic cells. In Vitro 6:162–172; 1970.
Falkmer, S.; Emdin, S.; Have, N., et al. Insulin in invertebrate and cyclostomes. Am. Zool. 13:625–638; 1973.
Furst, A.; Mahowald, A. P. Differentiation of primary embryonic neuroblasts in purified neural cell cultures fromDrosophila. Dev. Biol. 109:184–192; 1985.
Fujimori, K.; Yamamoto, K. Determination of acetylcholine and choline in perchlorate extracts of brain tissue using liquid chromatography-electrochemistry with an immobilized-enzyme reactor. J. Chromatogr. 414:167–173; 1987.
Garofalo, R. S.; Rosen, O. M. Tissue localization ofDrosophila melanogaster insulin receptor transcripts during development. Mol. Cell. Biol. 8:1638–1647; 1988.
Gorczyca, M.; Hall, J. C. Immunohistochemical localization of choline acetyltransferase during development and inCha ts mutants ofDrosophila melanogaster. J. Neurosci. 7:1361–1369; 1987.
Jan, L. Y.; Jan, Y. N. Antibodies to horseradish peroxidase as specific neuronal markers inDrosophila and in grasshopper embryos. Proc. Natl. Acad. Sci. USA 79:2700–2704; 1982.
Katz, F.; Moats, W.; Jan, L. N. A carbohydrate epitope expressed uniquely on the cell surface ofDrosophila neurons is altered in the mutantnac (neurally altered carbohydrate). EMBO J. 7:3471–3477; 1988.
Kim, Y-T.; Wu, C-F. Distinctions in growth cone morphology and motility between monopolar and multipolar neurons inDrosophila CNS cultures. J. Neurobiol. 22:263–275; 1991.
Kim Y-T.; Wu, C-F. Reversible blockage of neurite development and growth cone formation in neuronal cultures of a temperature-sensitive mutant ofDrosophila. J. Neurosci. 7:3245–3255; 1987.
Kojima, T.; Sone, M.; Michiue, T.; Saigo, K. Mechanism of induction ofbar-like eye malformation by transient overexpression ofbar homeobox gene inDrosophila melanogaster. Genetica, in press.
Kuroda, Y.; Tamura, S. A technique for the culture of melanotic tumors ofDrosophila melanogaster in the synthetic medium. Med. J. Osaka. Univ. 7:137–144; 1956.
Laemmli, U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685; 1970.
Lendahl, U.; McKsay, R. D. G. The use of cell lines in neurobiology. TINS 13:132–137; 1990.
Lewis, S. E.; Smallman, B. N. The estimation of acetylcholine in insects. J. Physiol. 134:241–256; 1956.
Meneses, P.; Ortiz, M. A. A protein extract fromDrosophila melanogaster with insulin-like activity. Comp. Biochem. Physiol. 51A:483–485; 1975.
Miyake, T.; Ueda, R. Establishment of embryonic cell lines inDrosophila melanogaster. Protein, Nucleic Acid Enzyme Suppl.: 314–318; 1984.
Peel, D. J.; Milner, M. J. The diversity of cell morphology in cloned cell lines derived fromDrosophila imaginal discs. Roux’s Arch. Dev. Biol. 198:479–482; 1990.
Saigo, K.; Ueda, R.; Miyake, T. Polymorphism and stability of histone gene clusters inDrosophila melanogaster cultured cells. Biochim. Biophys. Acta 740:390–401; 1983.
Salvaterra, P. M.; McCaman, R. E. Choline acetyltransferase and acetylcholine levels inDrosophila melanogaster. A study using two temperature-sensitive mutants. J. Neurosci. 5:903–910; 1985.
Schaeffer, W. I. Terminology associated with cell, tissue and organ culture, molecular biology and molecular genetics. In Vitro. Cell. Dev. Biol. 26:97–101; 1990.
Schneider, I. Cell lines derived from late embryonic stages ofDrosophila melanogaster. J. Embryol. Exp. Morph. 27:353–365; 1972.
Seecof, R. L.; Alleaume, M.; Teplitz, R. L., et al. Differentiation of neurons and myocytes in cell cultures made fromDrosophila gastrulae. Exp. Cell Res. 69:161–173; 1971.
Seecof, R.; Dewhurst, S. Insulin is aDrosophila hormone and acts to enhance the differentiation of embryonicDrosophila cells. Cell Differ. 3:63–70; 1974.
Shields, G.; Sang, J. H. Characteristics of five cell types appearing duringin vitro culture of embryonic material fromDrosophila melanogaster. J. Embryol. Exp. Morph. 23a:53–69; 1970.
Snow, P. M.; Patel, N. H.; Harrelson, A., et al. Neural-specific carbohydrate moiety shared by many surface glycoproteins inDrosophila and grasshopper embryos. J. Neurosci. 7:4137–4144; 1987.
Straus, D. S. Effects of insulin on cellular growth and proliferation. Life Sci. 29:2131–2139; 1981.
Ui, K.; Ueda, R.; Miyake, T. Cell lines from imaginal discs ofDrosophila melanogaster. In Vitro Cell. Dev. Biol. 23:707–710; 1987.
Ui, K.; Ueda, R.; Miyake, T.In vitro cultures of cells from different kinds of imaginal discs ofDrosophila melanogaster. Jpn. J. Genet. 63:33–41; 1988.
Ui, K.; Togashi, S.; Ueda, R., et al. Establishment of cell lines from larval central nervous system ofDrosophila melanogaster. Jpn. J. Genet. 63:606; 1988.
Ui, K.; Ueda, R.; Miyake, T.In vitro culture of cells from dissociated imaginal disc ofDrosophila melanogaster. In: Mitsuhashi, J., ed. Invertebrate cell system applications, vol. II. Florida: CRC press; 1989:212–231.
Ui, K.; Sakuma, M.; Nishihara, S., et al. Neurotransmitter analysis in cell lines from larval CNS ofDrosophila melanogaster. Jpn. J. Genet. 64:492; 1989.
White, K.; Kankel, D. R. Patterns of cell division and cell movement in the formation of the imaginal nervous system inDrosophila melanogaster. Dev. Biol. 65:296–321; 1978.
Wu, C-F. Neurogenetic studies ofDrosophila central nervous system neurons in culture. In: Beadle, D. J.; Lees, G.; Kater, S. B., eds. Cell culture approaches to invertebrate neuroscience. New York: Academic Press; 1988:149–187.
Wu, C-F.; Sakai, K.; Saito, M., et al. GiantDrosophila neurons differentiated from cytokinesis-arrested embryonic neuroblasts. J. Neurobiol. 21:499–507; 1989.
Wu, C-F,; Suzuki, N.; Poo, M-M. Dissociated neurons from normal and mutantDrosophila larval nervous system in cell culture. J. Neurosci. 3:1888–1899; 1983.
Wyss, C. Ecdysterone, insulin and fly extract needed for the proliferation of normalDrosophila cells in defined medium. Exp. Cell Res. 139:297–307; 1982.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ui, K., Nishihara, S., Sakuma, M. et al. Newly established cell lines fromDrosophila larval CNS express neural specific characteristics. In Vitro Cell Dev Biol - Animal 30, 209–216 (1994). https://doi.org/10.1007/BF02632042
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02632042