Skip to main content
Log in

The morphological response of Kc-H cells to ecdysteroids: Hormonal specificity

  • Published:
Wilhelm Roux's archives of developmental biology Aims and scope Submit manuscript

Summary

Cells of the line Kc, derived fromDrosophila melanogaster embryos, extend long processes when exposed to ecdysteroid hormones. We have devised a quantitative assay for this morphological response, using the subline Kc-H. The assay was used to characterize the conditions required for the response. A halfmaximal response is elicited by approximately 10−8M 20-hydroxyecdysone; the response is saturated by 10−7M 20-hydroxyecdysone, which causes detectable elongation within a few hours, and a maximal response after 2–3 days. The response occurs substantially normally in the absence of serum, during growth in suspension, and in over-crowded cultures. It is not elicited by cyclic nucleotides, vertebrate growth factors, or a variety of other non-ecdysteroid reagents. Of 60 ecdysteroid compounds tested, only those which were active in other insect test systems elicited the response, and the concentrations required were approximately proportional to the concentrations active in other in vitro systems. We conclude that the response of Kc cells to 20-hydroxyecdysone retains basic features of the ecdysteroid response of intact tissues and therefore that Kc cells are a useful model system for studying ecdysteroid action.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Agui N (1973) Quantitative bioassay of moulting hormone in vitro. Appl Entomol Zool 8:236–239

    Google Scholar 

  • Agui N (1976) Attempts to establish insect endocrine system in vitro. In: Maramorosch K (ed) Invertebrate tissue culture. Academic Press, New York, pp 133–160

    Google Scholar 

  • Agui N, Fukaya M (1973) Effects of moulting hormones and prothoracic glands on the development of wing discs of the cabbage armyworm (Mamestra brassicae L.) in vitro (Lepidoptera: Noctuidae). Appl Entomol Zool 8:73–82

    Google Scholar 

  • Ashburner M (1971) Induction of puffs in polytene chromosomes of in vitro cultured salivary glands ofDrosophila melanogaster. Nature New Biol 230:222–223

    Google Scholar 

  • Ashburner M (1973) Sequential gene activation by ecdysone in polytene chromosomes ofDrosophila melanogaster. I. Dependence upon ecdysone concentration. Dev Biol 35:47–61

    Google Scholar 

  • Ashburner M, Richards G (1976) The role of ecdysone in the control of gene activity in the polytene chromosomes ofDrosophila. In: Lawrence PA (ed) Insect development. Wiley, New York, pp 203–225

    Google Scholar 

  • Berger E, Frank M, Abell MC (1980) Ecdysone-induced changes in protein synthesis in embryonicDrosophila cells in culture. In: Kurstak E, Maramorosch K, Dübendorfer A (eds) Invertebrate systems in vitro. Elsevier/North Holland, Amsterdam, pp 195–208

    Google Scholar 

  • Berger E, Ringler R, Alahiotis S, Frank M (1978) Ecdysone-induced changes in morphology and protein synthesis inDrosophila cell cultures. Dev Biol 62:498–511

    Google Scholar 

  • Best-Belpomme M, Courgeon A-M (1977) Ecdysterone and acetylcholinesterase activity in culturedDrosophila cells. FEBS Lett 82:345–347

    Google Scholar 

  • Best-Belpomme M, Courgeon A-M, Echalier G (1980) Development of a model for the study of ecdysteroid action:Drosophila melanogaster cells established in vitro. In: Hoffmann J (ed), Progress in ecdysone research. Elsevier/North Holland, Amsterdam, in press

    Google Scholar 

  • Best-Belpomme M, Courgeon A-M, Rambach A (1978) β-Galactosidase is induced by hormone inDrosophila melanogaster cell cultures. Proc Natl Acad Sci USA 75:6102–6106

    Google Scholar 

  • Bliss CI (1970) Statistics in biology, vol 2. McGraw-Hill, New York, p 172

    Google Scholar 

  • Bollenbacher WE, Galbraith MN, Gilbert LI, Horn DHS (1977) In vitro metabolism of 3β-hydroxy-, and 3β, 14α-dihydroxy-[3α-3H]-5β-cholest-7-en-6-one by the prothoracic glands ofManduca sexta. Steroids 29:47–63

    Google Scholar 

  • Borst DW, Bollenbacher WE, O'Connor JD, King DS, Fristrom JW (1974) Ecdysone levels during metamorphosis ofDrosophila melanogaster. Dev Biol 39:308–316

    Google Scholar 

  • Boyd M, Ashburner M (1977) The hormonal control of salivary gland secretion inDrosophila melanogaster: Studies in vitro. J Insect Physiol 23:517–523

    Google Scholar 

  • Bullière F, Bullière D (1977) Régénération, différenciation et hormones de mues chez l'embryon de blatte en culture in vitro. Wilhelm Roux's Archives 182:255–275

    Google Scholar 

  • Canonica L, Danieli B, Weisza-Vincze I, Ferrari G (1972) Structure of muristerone A, a new phytoecdysone. Chem Commun 1972:1060–1061

    Google Scholar 

  • Caruelle JP, Cassier P, Joulie-Delorme C (1978) Dose-effect relationships of α- and β-ecdysone on cuticular deposition and cellular alterations of larval epidermis ofSchistocerca gregaria in vitro. Ann Sci Nat Zool Biol Anim 20:53–78

    Google Scholar 

  • Cherbas L, Cherbas P (1970) Distribution and metabolism of α-ecdysone in pupae of the silkworm,Antheraea polyphemus. Biol Bull 138:115–128

    Google Scholar 

  • Cherbas L, Cherbas P, Savakis C, Demetri G, Manteuffel-Cymborowska M, Yonge CD, Williams CM (1980) Studies of ecdysteroid action on aDrosophila cell line. In: Kurstak E, Maramorosch K, Dübendorfer A (eds) Invertebrate systems in vitro. Elsevier/North Holland, Amsterdam, pp 217–228

    Google Scholar 

  • Cherbas P, Cherbas L, Williams CM (1977) Induction of acetylcholinesterase activity by β-ecdysone in aDrosophila cell line. Science 197:275–277

    Google Scholar 

  • Cherbas P, Cherbas L, Demetri G, Manteuffel-Cymborowska M, Savakis C, Yonge CD, Williams CM (1980) Ecdysteroid hormone effects on aDrosophila cell line. In: Roy AK, Clark JH (eds), Gene regulation by steroid hormones. Springer, Berlin Heidelberg New York, pp 278–308

    Google Scholar 

  • Chihara CJ, Petri WH, Fristrom JW, King DS (1972) The assay of ecdysones and juvenile hormones onDrosophila imaginal disks in vitro. J Insect Physiol 18:1115–1123

    Google Scholar 

  • Chong YK, Galbraith MN, Horn DHS (1970) Isolation of deoxycrustecdysone, deoxyecdysone, and α-ecdysone from the fernBlechnum minus. Chem Commun 1970:1217–1218

    Google Scholar 

  • Courgeon A-M (1972a) Action of insect hormones at the cellular level. Morphological changes of a diploid cell line ofDrosophila melanogaster treated with ecdysone and several analogues in vitro. Exp Cell Res 74:327–336

    Google Scholar 

  • Courgeon A-M (1972b) Effects of α- and β-ecdysone on in vitro diploid cell multiplication inDrosophila melanogaster. Nature New Biol 238:250–251

    Google Scholar 

  • Courgeon A-M (1975) Action conjuguée de l'hormone juvénile et de l'ecdysterone sur des lignées cellulaires de Drosophile in vitro. CR Acad Sci Paris Ser D 280:2563–2566

    Google Scholar 

  • Doane WW (1973) Role of hormones in insect development. In: Counce SJ, Waddington CH (eds), Developmental systems: Insects, vol 2. Academic Press, New York, pp 291–497

    Google Scholar 

  • Dübendorfer A, Shields G, Sang JH (1975) Development and differentiation in vitro ofDrosophila imaginal disc cells from dissociated early embryos. J Embryol Exp Morphol 33:487–498

    Google Scholar 

  • Fain MJ, Riddiford LM (1977) Requirements for molting of the crochet epidermis of the tobacco hornworm larva in vivo and in vitro. Wilhelm Roux's Archives 181:285–307

    Google Scholar 

  • Fristrom JH, Yund MA (1976) Characteristics of the action of ecdysones onDrosophila imaginal discs cultured in vitro. In: Maramorosch K (ed) Invertebrate tissue culture, Academic Press, New York, pp 161–178

    Google Scholar 

  • Fukushima T, Yagi S (1975) Hormonal effect on cultivated insect tissues. III. Effects of α- and β-ecdysones or prothoracic glands on spermiogenesis in two noctuid moths in vitro (Lepidoptera: Noctuidae). Appl Entomol Zool 10:220–225

    Google Scholar 

  • Galbraith MN, Horn DHS, Middleton EJ, Thomson JA (1973) Biological activity of synthetic molting hormone analogues in the blowflyCalliphora stygia. Experientia 29:19

    Google Scholar 

  • Galbraith MN, Horn DHS, Porter QN, Hackney RJ (1968) Structure of podecdysone A, a steroid with moulting hormone activity from the bark ofPodocarpus elatus R. Br. Chem Commun 1968:971–972

    Google Scholar 

  • Goodwin TW, Horn DHS, Karlson P, Koolman J, Nakanishi K, Robbins WE, Siddall JB, Takemoto T (1978) Ecdysteroids: A new generic term. Nature 272:122

    Google Scholar 

  • Gvozdev VA, Kakpakov VT, Mukhovatova LM, Polukarova LG, Tarantul VZ (1974) Effect of ecdysterone on cell growth and macromolecular synthesis in established embryonic cell lines ofDrosophila melanogaster. Ontogenez 5:33–42

    Google Scholar 

  • Hetru C, Lagueux M, Bang L, Hoffmann JA (1978) Adult ovaries ofLocusta migratoria contain the sequence of biosynthetic intermediates for ecdysone. Life Sci. 22:2141–2154

    Google Scholar 

  • Hikino H, Hikino Y (1970) Arthropod molting hormones. Fortschr Chem Org Naturst 28:256–312

    Google Scholar 

  • Hikino H, Hikino Y, Nomoto K, Takemoto T (1968) Cyasterone, an insect metamorphosing substance fromCyathula capitata: Structure. Tetrahedron 24:4895–4906

    Google Scholar 

  • Hodgetts RB, Sage B, O'Connor JD (1977) Ecdysone titers during postembryonic development ofDrosophila melanogaster. Dev Biol 60:310–317

    Google Scholar 

  • Hoffmeister F, Nakanishi K, Koreeda M, Hsu HY (1968) The moulting hormone activity of ponasterones in theCalliphora test. J Insect Physiol 14:53–54

    Google Scholar 

  • Horn DHS (1971) The ecdysones. In: Jacobson M, Crosby DG (eds) Naturally occurring insecticides. Dekker, New York, pp 333–459

    Google Scholar 

  • Imai S, Hori M, Fujioka S, Murata E, Goto M, Nakanishi K (1968a) Isolation of four new phytoecdysones, makisterone A, B, C, D, and the structure of makisterone A, a C28 steroid. Tetrahedron Lett 1968:3883–3886

    Google Scholar 

  • Imai S, Fujioka S, Murata E, Sasakawa Y, Nakanishi K (1968b) The structures of three additional phytoecdysones fromPodocarpus macrophyllus, makisterone B, C, and D. Tetrahedron Lett 1968:3887–3890

    Google Scholar 

  • Imai S, Toyosato T, Sakai M, Sato Y, Fujioka S, Murata E, Goto M (1969) Isolation of cyasterone and ecdysterone from plant materials. Chem Pharm Bull Jpn 17:340–342

    Google Scholar 

  • Jizba J, Herout V, Šorm F (1967) Polypodine B—a novel ecdysonelike substance from plant material. Tetrahedron Lett 1967:5139–5143

    Google Scholar 

  • Kimura S, Kobayashi M (1975) Prothoracicotropic action of ecdysone analogues inBombyx mori. J Insect Physiol 21:417–421

    Google Scholar 

  • King DS, Siddall JB (1969) Conversion of α-ecdysone to β-ecdysone by crustaceans and insects. Nature 221:955–956

    Google Scholar 

  • Kuroda Y (1969) Growth and differentiation of embryonic cells ofDrosophila melanogaster in vitro. Jpn J Genet [Suppl 1] 44:42–50

    Google Scholar 

  • Landureau JC (1976) Insect cell and tissue culture as a tool for developmental biology. In: Kurstak E, Maramorosch K (eds), Invertebrate tissue culture. Academic Press, New York, pp 101–130

    Google Scholar 

  • Laverdure A-M (1976) Recherche des conditions nécéssaires à l'évolution de l'ovaire de la nymphe deTenebrio molitor. Etude en culture in vitro. Bull Soc Zool Fr [Suppl 2] 101:8–11

    Google Scholar 

  • Lee E, Liu Y-T, Solomon PH, Nakanishi K (1976) Stereospecific conversion of diosgenin to α-ecdysone. J Am Chem Soc 83:1634–1635

    Google Scholar 

  • Marks EP (1973) Deposition of insect cuticle in vitro: Differential responses to α- and β-ecdysone. Gen Comp Endocrinol 21:472–477

    Google Scholar 

  • Maroy P, Dennis R, Beckers C, Sage BA, O'Connor JD (1978) Demonstration of an ecdysteroid receptor in a cultured cell line ofDrosophila melanogaster. Proc Natl Acad Sci USA 75:6035–6038

    Google Scholar 

  • Milner MJ, Haynie JL (1979) Fusion ofDrosophila eye-antennal imaginal discs during differentiation in vitro. Wilhelm Roux's Archives 185:363–370

    Google Scholar 

  • Milner MJ, Sang JH (1974) Relative activities of α-ecdysone and β-ecdysone for the differentiation in vitro ofDrosophila melanogaster imaginal discs. Cell 3:141–143

    Google Scholar 

  • Mitsuhashi J, Grace TDC (1970) The effects of insect hormones on the multiplication rates of cultured insect cells in vitro. Appl Entomol Zool 5:182–188

    Google Scholar 

  • Mitsui T, Riddiford LM (1976) Pupal cuticle formation byManduca sexta epidermis in vitro: Patterns of ecdysone sensitivity. Dev Biol 54:172–186

    Google Scholar 

  • Morisaki M, Awata N, Fujimoto Y, Ikekawa N (1975) Steroidal allenes as inhibitors of sterol biosynthesis. Chem Commun 1975:362–363

    Google Scholar 

  • Moriyama H, Nakanishi K, King DS, Okauchi T, Siddall JB, Hafferl W (1970) On the origin and metabolic fate of α-ecdysone in insects. Gen Comp Endocrinol 15:80–87

    Google Scholar 

  • Mukhovatova LM, Kakpakov VT (1975) Effect of α- and β-ecdysone on the differentiation ofDrosophila melanogaster imaginal discs cultivated in vitro. Ontogenez 6:80–87

    Google Scholar 

  • Oberlander H (1969) Effects of ecdysone, ecdysterone, and inokosterone on the in vitro initiation of metamorphosis of wing disks ofGalleria mellonella. J Insect Physiol 15:297–304

    Google Scholar 

  • Oberlander H (1976) Dissociation and reaggregation of fat body cells during insect metamorphosis. In: Kurstak E, Maramorosch K (eds) Invertebrate tissue culture. Academic Press, New York, pp 241–246

    Google Scholar 

  • Oberlander H, Ferkovich SM, Van Essen F, Leach CE (1978) Chitin biosynthesis in imaginal discs cultured in vitro. Wilhelm Roux's Archives 185:95–98

    Google Scholar 

  • Oberlander H, Leach CE (1975) Uptake and incorporation of D-glucosamine-6-3H into imaginal disks: Effects of alpha-ecdysone and beta-ecdysone in vitro. Insect Biochem 5:235–242

    Google Scholar 

  • Oberlander H, Leach CE, Tomblin C (1973) Cuticle deposition in imaginal disks of three species of Lepidoptera: Effects of ecdysones in vitro. J Insect Physiol 19:993–998

    Google Scholar 

  • O'Connor JD, Maroy P, Beckers C, Dennis R, Alvarez CM, Sage BA (1980) Ecdysteroid receptors in culturedDrosophila cells. In: Roy AK, Clark JH (eds) Gene regulation by steroid hormones. Springer, Berlin Heidelberg New York, pp 263–277

    Google Scholar 

  • Ohmori K (1974) Effect of ecdysterone contact period on the development of wing disks of the fleshfly,Sarcophaga peregrina, in vitro. J Insect Physiol 20:1697–1706

    Google Scholar 

  • Ohmori K, Ohtaki T (1973) Effects of ecdysone analogues on development and metabolic activity of wing disks of the fleshfly,Sarcophaga peregrina, in vitro. J Insect Physiol 19:1199–1210

    Google Scholar 

  • Ohtaki T, Milkman RD, Williams CM (1967) Ecdysone and ecdysone analogues: Their assay on the fleshflySarcophaga peregrina. Proc Natl Acad Sci USA 58:981–984

    Google Scholar 

  • Richards G (1978) Sequential gene activation by ecdysone in polytene chromosomes ofDrosophila melanogaster. VI. Inhibition by juvenile hormone. Dev Biol 66:32–42

    Google Scholar 

  • Riddiford LM (1976) In vitro action of ecdysone and juvenile hormone on epidermal commitment in the tobacco hornworm. In: Kurstak E, Maramorosch K (eds) Invertebrate tissue culture. Academic Press, New York, pp 213–222

    Google Scholar 

  • Riddiford LM (1978) Ecdysone-induced change in cellular commitment of the epidermis of the tobacco hornworm,Manduca sexta, at the initiation of metamorphosis. Gen Comp. Endocrinol 34:438–446

    Google Scholar 

  • Rimpler H (1969) Pterosteron, Polypodin B, und ein neues ecdysonartiges Steroid (Viticosteron E) ausVitex megapotamica (Verbenaceae). Tetrahedron Lett 1969:329–333

    Google Scholar 

  • Robb JA (1969) Maintenance of imaginal discs ofDrosophila melanogaster in chemically defined media. J Cell Biol 41:876–885

    Google Scholar 

  • Robbins WE, Kaplanis JN, Thompson MJ, Shortino TJ, Cohen CF, Joyner SC (1968) Ecdysones and analogs: Effects on development and reproduction in insects. Science 161:1158–1160

    Google Scholar 

  • Robbins WE, Kaplanis JN, Thompson MJ, Shortino TJ, Joyner SC (1970) Ecdysones and synthetic analogs: Molting hormone activity and inhibitive effects on insect growth, metamorphosis, and reproduction. Steroids 16:105–125

    Google Scholar 

  • Robertson J, Pipa R (1973) Metamorphic shortening of interganglionic connectives ofGalleria mellonella (Lepidoptera) in vitro: Stimulation by ecdysone analogs. J Insect Physiol 19:673–679

    Google Scholar 

  • Rosset R (1978) Effects of ecdysone on aDrosophila cell line. Exp Cell Res 111:31–36

    Google Scholar 

  • Sláma K, Romanuk M, Šorm F (1974) Insect hormones and bioanalogues. Springer, Berlin Heidelberg New York

    Google Scholar 

  • Solomon F (1979) Detailed neurite morphologies of sister neuroblastoma cells are related. Cell 16:165–169

    Google Scholar 

  • Svoboda JA, Thompson MJ, Robbins WE, Kaplanis JN (1978) Insect steroid metabolism. Lipids 13:742–753

    Google Scholar 

  • Szöllösi A, Landureau J-C (1977) Imaginal cell differentiation in the spermiduct ofSamia cynthia (Lepidoptera): Responses in vitro to ecdysone and ecdysterone. Biol Cell 28:23–36

    Google Scholar 

  • Takeda N (1972) Effect of some phytoecdysones on the spermatogenesis of the slug moth prepupa,Monema flavescens Walker (Lepidoptera: Heterogeneidae). Appl Entomol Zool 7:37–39

    Google Scholar 

  • Takemoto T, Ogawa S, Nishimoto N, Mue K (1967) Studies on the consituents ofAchyranthis radix. V. Insect hormone activity of ecdysterone and inokosterone on the flies. Yakagaku Zasshi 87:1481–1483

    Google Scholar 

  • Thompson MJ, Robbins WE, Cohen CF, Kaplanis JN, Dutky SR, Hutchins RFN (1971) Synthesis and biological activity of 5β-hydroxy analogs of α-ecdysone. Steroids 17:399–409

    Google Scholar 

  • Wyatt GR (1972) Insect hormones. In: Litwack G (ed) Biochemical actions of hormones, vol 2. Academic Press, New York, pp 385–490

    Google Scholar 

  • Wyatt SS, Wyatt GR (1971) Stimulation of RNA and protein synthesis in silkmoth pupal wing tissue by ecdysone in vitro. Gen Comp Endocrinol 16:369–374

    Google Scholar 

  • Wyss C (1976) Juvenile hormone analogue counteracts growth stimulation and inhibition by ecdysones in clonalDrosophila cell line. Experientia 32:1272–1274

    Google Scholar 

  • Wyss C (1980) Cell hybrid analysis of ecdysone sensitivity and resistance inDrosophila cell lines. In: Kurstak E, Maramorosch K, Dübendorfer (eds) Invertebrate systems in vitro. Elsevier/North Holland, Amsterdam, in press

    Google Scholar 

  • Yund MA, King DS, Fristrom JW (1978) Ecdysteroid receptors in imaginal discs ofDrosophila melanogaster. Proc Natl Acad Sci USA 75:6039–6043

    Google Scholar 

  • Zdarek J, Denlinger DL (1975) Action of ecdysoids, juvenoids, and non-hormonal agents on termination of pupal diapause in the fleshfly. J Insect Physiol 21:1193–1202

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cherbas, L., Yonge, C.D., Cherbas, P. et al. The morphological response of Kc-H cells to ecdysteroids: Hormonal specificity. Wilhelm Roux' Archiv 189, 1–15 (1980). https://doi.org/10.1007/BF00848562

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00848562

Key words

Navigation