Use of Microderivatization Techniques in Combination with Thin-Layer Chromatography, Liquid Chromatography and Gas Chromatography-Mass Spectrometry for Investigation of Juvenile Hormones and Related Compounds
The qualitative juvenile hormone (JH) profile in lepidopteran species, as determined by both in vivo and in vitro techniques, is markedly different from that in insects from other orders, the majority of which contain only JH III. A gas chromatographic-mass spectrometric (GC-MS) method for quantification of JHs in vivo was adapted to study the mode of JH biosynthesis by corpora allata incubated in vitro with selected 14C-radiolabeled substrates of high specific activity. We analyzed derivatives of JH using selected ion monitoring to measure ratios of the fragment ion and corresponding ions 2,4, and 6 mass units higher, allowing determination of dilution of the propionate unit which is incorporated as an intermediate. The results showed conclusively that the ethyl branch of JH II from Manduca sexta derives ultimately from metabolism of the branched-chain amino acids isoleucine and valine. A purification procedure for isolation of JHs from biological tissue was modified to allow isolation of JH acids and compounds structurally related to JH, including JH diols, methyl farnesoate (MF), farnesoic acid (FA), and farnesol. To allow analysis, JH acids were methylated and the resulting JHs were detected and quantified, as their d3-methoxyhydrin derivatives, by GC-MS with selected ion monitoring (SIM). A detailed study of fifth stadium Manduca sexta showed that JH I and II acids predominate over the corresponding esters at the prepupal stage. GC-MS methods were devised for quantification of JH diols, MF, FA, and farnesol from arthropods. None of the latter compounds, nor any of the known JHs, could be detected in the hemipteran Oncopeltus fasciatus. Ring glands from a few dipteran species appear to secrete a new JH in vitro, and details are given for the isolation and analysis of this compound. Haemolymph from several crustaceans contain detectable levels of MF, and mandibular organs from crustaceans secrete MF and/or FA in vitro.
Unable to display preview. Download preview PDF.
- 2.F. C. Baker, Techniques for identification and quantification of juvenile hormones and related compounds in arthropods, in: “Morphogenese Hormones of Arthropods/Recent Advances in Comparative Morphology, Physiology and Development of Arthropods”, A. P. Gupta, ed., Rutgers University Press, New Brunswick, pp. 389–453, in press (1990).Google Scholar
- 3.D. A. Schooley, F. C. Baker, L. W. Tsai, C. A. Miller and G. C. Jamieson, Juvenile hormones, 0, I and II exist only in Lepidoptera, in: “Biosynthesis, Metabolism and Mode of Action of Invertebrate Hormones”, J. Hoffmann and M. Porchet, eds., Springer-Verlag, Berlin, pp. 373–383 (1984).CrossRefGoogle Scholar
- 7.A. Dorn, Structure and function of the embryonic corpus allatum of Oncopeltus fasciatus Dallas (Insecta, Heteroptera), Verhandlungen der Deutschen Zoologischen Gesellschaft, pp. 85–89 (1975).Google Scholar
- 9.D. S. Richard, S. W. Applebaum, T. J. Sliter, F. C. Baker, D. A. Schooley, C. C. Reuter, V. C. Henrich and L. I. Gilbert, Juvenile hormone bisepoxide biosynthesis in vitro by the ring of Drosophila melanogaster. a putative juvenile hormone in the higher Diptera, Proc. Nat. Acad. Sci. USA, 86:1421–1425 (1989).CrossRefPubMedPubMedCentralGoogle Scholar
- 12.H. Laufer, M. Landau, E. Homola and D. W. Borst, Methyl farnesoate: its site of synthesis and regulation of secretion in a juvenile crustacean, Insect Biochem., 17:1129–1131(1987).Google Scholar
- 16.F. C. Baker and D. A. Schooley, Preparation of (10R.S)-[10–3HJjuvenile hormone III and (10R.S, 11S.R)-[10–3HUuvenile hormone O: conversion of [10–3H]juvenile hormone III to methyl (2E, 6E)-[10–3H]farnesoate and (2E,6E)-[10–3H]farnesol, J. Labeled Comp. Radiopharmaceut, 5:533–543 (1986).Google Scholar
- 19.P. A. Brindle, D. A. Schooley, L. W. Tsai and F. C. Baker, Comparative metabolism of branched-chain amino acids to precursors of juvenile hormone biogenesis in corpora allata of lepidopterous versus non-lepidopterous insects, J. Biol Chem., 263:10653–10657(1988).Google Scholar
- 24.K. H. Trautmann, P. Masner, A. Schüler, M. Suchy and H.-K. Wipf, Evidence of the juvenile hormone methyl(2E,6E)-10,1 l-epoxy-3,7,1 l-trimethyl-2,6-dodecadieno-ate JH-3) in insects of four orders, Zeitschrift fur Naturforschung., 29c:757–759 (1974).Google Scholar
- 26.F. C. Baker, C. A. Miller, L. W. Tsai, G. C. Jamieson, D. C. Cerf and D. A. Schooley, The effect of juvenoids, anti juvenile hormone agents, and several intermediates of juvenile hormone biosynthesis on the in vivo juvenile hormone levels in Manduca sexta larvae, Insect Biochem., 16:741–747 (1986).CrossRefGoogle Scholar
- 27.P. O. Lawrence, F. C. Baker, L. W. Tsai, C. A. Miller, D. A. Schooley and L. Geddes, JH III levels in superparasitized pharate pupae of Anastrepha suspensa (Diptera: tephritidae) and in larvae of the parasitic wasp Biosteres longicaudatus (Hymenoptera: braconidae), Arch, Insect Biochem. Physiol., in press (1990).Google Scholar
- 28.G. Jones, T. Hanzlik, B. D. Hammock, D. A. Schooley, C. A. Miller, L. W. Tsai and F. C. Baker, The juvenile hormone titer during the penultimate and ultimate larval stadia of Trichoplusia ni, J. Insect Physiol., in press (1990).Google Scholar
- 32.W. S. Bowers, P. A. Marsella and P. H. Evans, Identification of an hemipteran juvenile hormone: in vitro biosynthesis of JH III by Dysdercus fasciatus, J. Exp. Zool., 228:555–559(1983).Google Scholar
- 35.B. Lanzrein, H. Imboden, C. Bürgin, E. Brüning and H. Gfeller, On titers, origin and functions of juvenile hormone III, methyl farnesoate, and ecdysteroids in embryonic development of the ovoviviparous cockroach Nauphoeta cinerea, in: “Biosynthesis, Metaboism and Mode of Action of Invertebrate Hormones”, J. Hoffmann and M. Porchet, eds., Springer-Verlag, Berlin, pp. 454–465 (1984).CrossRefGoogle Scholar