Characterization of Insulin-Like Peptides and Their Relation to Molt and Metabolism of the Shrimp Litopenaeus vannamei
Resumen
Insulin-like peptides are key regulators of metabolism, reproduction, and senescence in higher eukaryotic organisms.
They have been described in different invertebrates, including nematodes, molluscs, and insects. Insulin-like peptides
have also been detected in some decapod crustaceans, where they may display typical insulin effects. On the basis
that common biological function may be related to a similar structure, we searched for an insulin-related substance in
the shrimp Litopenaeus vannamei and further investigated its function. An IGF-I-like molecule was detected in
neurosecretory cells of the brain and thoracic ganglion in shrimps at different molt stages. Furthermore, we
demonstrated that ILP did not differentially stimulate ecdysteroid secretion as it is observed in some insect species.
An in vivo assay was performed and the results revealed that injections of heterologous insulin/IGF-I increased the
glycogen contents of gills and digestive gland from intermolt animals (fasting state), suggesting that an endogenous
ILP may display a conserved function in decapods. Moreover, cloning and sequencing of a cDNA from hemocytes
revealed that the sequence code for a precursor similar to the Insulin-like Growth Factor binding proteins. A
quantitative real-time RT-PCR study revealed the presence of LivIGFBP-related peptide transcript in a variety of
tissues, including nervous tissues, gills, and muscle, brain, thoracic ganglion and nerve cord. In addition, the
expression of this transcript differs according to the animal’s size, suggesting a possible role in growth regulation.
Descargas
Citas
Argue, B.J., Arce, S.M., Lotz, J.M., Moss, S.M. 2002. Selective breeding of Pacific white shrimp (Litopenaeus
vannamei) for growth and resistance to Taura Syndrome Virus. Aquaculture 204, 447-460.
Auernhammer, C.J., Fottner, C., Engelhardt, D., Bidlingmaier, M, Strasburger, C.J. Weber, M.M. 2002. Differential
regulation of insulin-like growth factor- (IGF) I and IGF-binding protein (IGFBP) secretion by human
peripheral blood mononuclear cells. Hormone Research 57, 15-21.
Badisco, L., Claeys, I., Van Hiel, M., Clynen, E., Huybrechts, J., Vandermissen, T., Van Soest, S., Vanden Bosch,
L., Simonet, G., Vanden Broeck, J. 2008. Purification and characterization of an insulin-related peptide in
the desert locust, Schistocerca gregaria: immunolocalization, cDNA cloning, transcript profiling and
interaction with neuroparsin. Journal of Molelucar Endocrinology 40, 137-150.
Badisco, L., Claeys, I., Van Loy, T., Van Hiel, M., Franssens, V., Simonet, G., Vanden Broeck, J. 2007.
Neuroparsins, a family of conserved arthropods neuropeptides. General and Comparative Endocrinology
, 64-71.
Bathgate, R.A., Samuel, C.S., Burazin, T.C., Layfield, S., Claasz, A.A., Reytomas, I.G., Dawson, N.F., Zhao, C.,
Bond, C., Summers, R.J., Parry, L.J., Wade, J.D., Tregear, G.W. 2002. Human relaxin gene 3 (H3) and the
equivalent mouse relaxin (M3) gene. Novel members of the relaxin peptide family. Journal of Biological
Chemistry 277, 1148-1157.
Baxter, R.C. 2000. Insulin-like growth factor (IGF) binding proteins: Interactions with IGFs and intrinsic
bioactivities. American Journal of Physiology 278, E967-E976.
Blais, C., Sefiani, M., Toullec, J-Y., Soyez, D. 1994. In vitro production of ecdysteroids by Y-organs of Penaeus
vannamei (Crustacea, Decapoda). Correlation with hemolymph titers. Invertebrate and Reproduction
Development 26, 3-12.
Burkhardt, E., Adham, I.M. Brosig, B., Gastmana, A., Mattei, M.G., Engel, W. 1994. Structural organization of the
porcine and human genes coding for a Leydig cell-specific insulin-like peptide (LEY I-L) and chromosomal
localization of the human gene (INSL3). Genomics 20, 13-19.
Cao C., Brown M.R. 2001. Localization of an insulin-like peptide in brains of two flies. Cell and Tissue Research
, 317-321.
Castille, F.L., Samocha, R.M., Lawrence, A.L., He, H., Frelier, P., Jaenike, F. 1993. Variability in growth and
survival of early postlarval shrimp (Penaeus vannamei Boone 1931). Aquaculture 113, 65-81.
Chang, E.S. 1995. Physiological and biochemical changes during the molt cycle in decapod crustaceans: an
overview. Journal of Experimental Marine Biology and Ecology 193, 1-14.
Chang, E.S., Tamone, S.L., Lin, W.-W., Oberbauer, A.M. 1995. Modifications to the paradigm of the hormonal
control of the crustacean molting: effects of metabolic inhibitors on the ecdysiotropic action of methyl
farnesoate and evidence for an insulin-like growth factor. In Proceedings of the international symposium on
biotechnology applications in aquaculture. Asian Fisheries Society 10, 183-195.
Chassin, D., Laurent, A., Janneuau, J.L., Berger, R., Bellet, D. 1995. Clonning of a new member of the insulin gene
superfamily (INSL4) expressed in human placenta. Genomics 29, 465-470.
Chuang, N-N., Wang, P-C. 1994. Characterization of insulin receptor from the muscle of the shrimp Penaeus
japonicus (crustacea: decapoda). Comparative Biochemical Physiology 108C, 289-297.
Claeys, I., Simonet, G., Poels, J., Van Loy, T., Vercammen, L., De Loof, A., Vanden Broeck, J. 2002. Insulin-related
peptides and their conserved signal transduction pathway. Peptides 23, 807-816.
Claeys, I., Simonet, G., Van Loy, T. De Loof, A., Vanden Broeck, J. 2003. cDNA cloning and transcript distribution
of two novel members of the neuroparsin family in the desert locust, Schistocerca gregaria. Insect
Molecular Biology 12, 473-481.
Clark, R. 1997. The somatogenic hormones and insulin-like growth factor-1: stimulators of lymphopoiesis and
immune function. Endocrine Reviews 18, 157-179.
Conklin, D., Lofton-Day, C.E., Haldeman, B.A., Ching, A., Whitmore, T.E., Lok, S., Jaspers, S. 1999. Identification
of INSL5, a new member of the insulin superfamily. Genomics 60, 50-56.
Davidson, J.K., Falkmer, S., Mehratra, B.K., Wilson, S., 1971. Insulin assay and light microscopial studies of
digestive organs in protostomian and deuterostomian species and coelenterates. General and Comparative
Endocrinology 17, 388-401.
Duret, L., Guex, N., Peitsch, M.C., Bairoch, A. 1998. New insulin-like proteins with atypical disulfide bond pattern
characterized in Caenorhabditis elegans by comparative sequence analysis and homology modelling.
Genome Research 8, 348-353.
Gallardo, N., Carrillo, O., Moltó, E., Deás, M., González-Suárez, R., Carrascosa JM, Ros M, Andrés A. 2003.
Isolation and biological characterization of a 6-kDa protein from hepatopancreas of lobster Panulirus argus
with insulin-like effects. General and Comparative Endocrinology 131: 284-290.
Gerisch, B., Weitzel, C., Kober-Eisermann, C., Rottiers, V., Antebi, A. 2001. A hormonal signaling pathway
influencing C. elegans metabolism, reproductive development, and life span. Developmental Cell 6, 841-
Goltzené, F., Holder, F., Charlet, M., Meister, M., Oka, T. 1992. Immunocytochemical localization of Bombyx-
PTTH-like molecules in neurosecretory cells of the brain of the migratory locust, Locusta migratoria. A
comparison with neuroparsin and insulin-related peptide. Cell and Tissue Research 269, 133-140.
Gross, P.S., Bartlett, T.C., Browdy, C.L., Chapman, R.W. and Warr, G.W. 2001. Immune gene discovery by
expressed sequence tag analysis of hemocytes and hepatopancreas in the Pacific White Shrimp, Litopenaeus
vannamei, and the Atlantic White Shrimp, L. setiferus. Developmental and Comparative Immunology 25,
-577.
Gutiérrez, A., Nieto, J., Pozo, F., Stern, S., Schoofs, L. 2007. Effects of insulin/IGF-I like peptides on glucose
metabolism in the white shrimp Penaeus vannamei. General and Comparative Endocrinology 153, 170-175.
Hansen, G.N., Hansen, B.L., Jorgensen, P.N., Scharrer, B. 1990. Immunocytochemical localization and
immunochemical characterization of an insulin-related peptide in the insect Leucophaea maderae. Cell and
Tissue Research 259, 265-273.
Hatt, P.J., Liebon, C., Moriniére, M., Oberlander, H., Porcheron, P. 1997. Activity of insulin growth factors and
shrimp neurosecretory organ extracts on a lepidopteran cell line. Archive of Insect Biochemistry and
Physiology 34, 313-328.
Honegger, B., Galic, M., Köhler, K., Wittwer, F., Brogiolo, W., Hafen, E., Stocker, H. 2008. Imp-L2, a putative
homolog of vertebrate IGF-binding protein 7, counteracts insulin signaling in Drosophila and is essential
for starvation resistance. Journal of Biology 7, 1-10.
Ikeya, T., Galic, M., Belawat, P., Nairz, K., Hafen, E. 2002. Nutrient-dependent expression of insulin-like peptides
from neuroendocrine cells in the CNS contributes to growth regulation in Drosophila. Current Biology 12,
-300.
Imai, Y., Moralez, A., Andag, U., Clarke, J.B. Busby, W.H. Clemmons, D.R. 2000. Substitutions for hydrophobic
amino acids in the N-terminal domains of IGFBP-3 and -5 markedly reduce IGF-I binding and alter their
biological actions. Journal of Biological Chemistry 275, 18188-18194.
Iwanaga, S. and Lee, B.L. 2005. Recent Advances in the innate immunity of invertebrate animals. Journal of
Biochemistry and Molecular Biology 38, 128-150.
Jones, J.I., Clemmons D.R. 1995. Insulin like growth factor and their binding proteins: Biologic actions. Endocrine
Reviews 16, 3-34.
Koman, A., Cazaubon, S., Couraud, P.O., Ullrich, A., Strosberg, A.D. 1996. Molecular characterization and in vitro
biological activity of placentin, a new member of the insulin gene family. Journal of Biological Chemistry
, 20238-20241.
Kooijman R., Scholtens L. E., Rijkers G. T. and Zegers B. J.M. 1995. Type I insulin-like growth factor receptor
expresion in different developmental stages of human thymocytes. Journal of Endocrinology 147, 203–209.
Kyte, J., Doolittle, R.F. 1982. A simple method for displaying the hydropathic character of a protein. Journal of
Molecular Biology 157, 105-132.
LeRoith, D., Shiloach, J., Roth, J., Lesniak, M.A. 1980. Evolutionary origins of vertebrate hormones: substances
similar to mammalian insulins are native to unicellular eukaryotes. Proceedings of the National Academy of
Sciences of the United States of America 77, 6184- 6188.
Lin, C.L., Wang, P.C., Chuang, N.N. 1993. Specific phosphorylation of membrane proteins of Mr 44,000 and Mr
,000 by the autophosphorylated insulin receptor from the hepatopancreas of the shrimp Penaeus monodon
(Crustacea: Decapoda). Journal of Experimental Zoology 267, 104-112.
Lok, S., Johnston, D.S., Conklin, D., Lofton-Day, C.E., Adams, R.L., Jelmberg, A.C., Whitmone, T.E., Schrader, S.,
Griswold, M.D., Jaspers, S.R. 2000. Identification of INSL6, a new member of the insulin family that is
expressed in the testis of the human and rat. Biology of Reproduction 62, 1593- 1599.
Manière G, Rondot I, Büllesbach EE, Gautron F, Vanhems E, Delbecque JP. 2004. Control of ovarian
steroidogenesis by insulin-like peptides in the blowfly (Phormia regina). Journal of Endocrinology 181:
-156.
Manor, R., Weil, S., Oren, S., Glazer, L., Aflalo, E.D., Ventura, T., Chalifa-Caspi, V., Lapidot, M., Sagi, A. 2007.
Insulin and gender: An insulin-like gene expressed exclusively in the androgenic gland of the male crayfish.
General and Comparative Endocrinology 150, 326-336.
McCusker, R.H. 1998. Controlling Insulin-like growth factor activity and the modulation of insulin-like growth
factor binding proteins and receptor binding. Journal of Diary Science 81, 1790-1800.
Mizoguchi, A., Ishizaki, H., Nagasawa, H., Kataoka, H., Isogai, A. 1987. A monoclonal-antibody against a synthetic
fragment of bombyxin (4K-prothoracicotropic hormone) from the silkmoth, Bombyx mori: characterization
and immunohistochemistry. Molecular and Cellular Endocrinology 51, 227-235.
Morrison, T.B., Weis, J.J., Wittwer, C.T. 1998. Quantification of low-copy transcripts by continuos SYBR Green I
monitoring during amplification. Biotechniques 24, 954-960.
Nagasawa H, Kataoka H, Isogai A, Tamura S, Suzuki A, Mizoguchi A, Fujiwara Y, Suzuki A, Takahashi SY,
Ishizaki H. 1986. Amino acid sequence of prothoracicotropic hormone of the silkworm Bombyx mori.
Proceedings of the National Academy of Sciences of the United States of America 83: 5840-5843.
Nogueira, B.V., Muehleisen, D.P., Whisenton, L.R., Gray, R.S., Bollenbacher, W.E. 1997. Life cycle expression of a
bombyxin-like neuropeptide in the tobacco hornworm, Manduca sexta. Journal of Insect Physiology 43, 47-
Nyman, T. and Pekonen, F. 1993. The expression of insulin-likegrowth factors and their binding proteins in normal
human lymphocytes. Acta Endocrinológica 128: 168–172.
Olinski, R.P., Lundin, L-G., Hallbook, F. 2006. Conserved synteny between the Ciona genome and human
paralogons identifies large duplication events in the molecular evolution of the insulin-relaxin gene family.
Molecular Biology and Evolution 23, 10-22.
Porcheron, P., Morinières, M., Grasi, J., Pradelles, P. 1989. Development of an enzyme immunoassay for
ecdysteroids using acetyl-cholinesterase as label. Insect Biochemistry 19, 117-122.
Reinecke, M., Betzler, D., Drankenberg, K., Falkmer, S., Sara, V.R. 1993. Occurrence of members of the insulin
superfamily in central nervous system and digestive tract of protochordates. Histochemistry 99, 277-285.
Reinecke, M., Drakenberg, K., Falkmer, S., Sara, V.R. 1991. Presence of IGF-1-like peptides in the neuroendocrine
system of the atlantic hagfish, Myxine glutinosa (cyclostomata): evidence derived by chromatography,
radioimmunoassay and immunohistochemistry. Histochemistry 96, 191-196.
Richardson, N.A., Anderson, A.J., Sara, V.R., 1997. The effect of insulin/IGF on glucose and leucine metabolism in
the redclaw crayfish (Cherax quadricarinatus). General and Comparative Endocrinology 105, 287-229.
Riehle, M.A., Fan, Y., Cao, C., Brown, M.R. 2006. Molecular characterization of insulin-like peptides in the yellow
fever mosquito, Aedes aegypti: Expression, cellular localization, and phylogeny. Peptides 27, 2547-2560.
Robertson, L., Bray, W., Leung-Trujillo, J., Lawrence, A. 1987. Practical molt staging of Penaeus setiferus and
Penaeus stylirostris. Journal of World Aquaculture Society 18, 180-185.
Sanders, B. 1983a. Insulin-like peptides in the lobster Homarus americanus. I. Insulin immunoreactivity. General
and Comparative Endocrinology 50, 366-373.
Sanders, B. 1983b. Insulin-like peptides in the lobster Homarus americanus. II Insulin-like Biological activity.
General and Comparative Endocrinology 50, 374-377.
Sevala, V.M., Sevala, V.L., Loughton, B.G. 1993. Insulin-like molecules in the beetle Tenebrio molitor. Cell and
Tissue Research 273, 71-77.
Shimasaki, S., Shimonaka, M., Zhang, H.P., Ling, N. 1991. Identification of five different insulin-like growth factor
binding proteins (IGFBPs) from adult rat serum and molecular cloning of a novel IGFBP in rat and human.
Journal of Biological Chemistry 266, 10646-10653.
Sitar, T., Popowicz, G.M., Siwanowicz, I., Huber, R., Holak, T.A. 2006. Structural basis for the inhibition of insulinlike
growth factors by insulin-like growth factor-binding proteins. Proceedings of the National Academy of
Sciences of the United States of America 103, 13028- 13033.
Smit, A.B., Van Kesteren, R.E., Li, K.W., Van Minnen, J., Spijker, S., Van Heerikhuizen, H., Geraerts, W.P.M.
Towards understanding the role of insulin in the brain: lessons from insulin-related signaling systems
in the invertebrate brain. Progress in Neurobiology 54, 35-54.
Söderhäll, K. and Cerenius, L. 1992. Crustacean immunity. Annual Review of Fish Diseases 2, 3-23.
Tatar, M., Bartke, A., Antebi, A. 2003. The endocrine regulation of aging by insulin-like signals. Science 299, 1346-
Tatar, M., Kopelman, A., Epstein, D., Tu, M.P., Garofalo, R.S. 2001. A mutant Drosophila insulin receptor homolog
that extends life-span and impairs neuroendocrine function. Science 292, 107-110.
Vafopoulou X, Steel CGH. 1997. Ecdysteroidogenic action of Bombyx prothoracicotropic hormone and bombyxin
on the prothoracic glands of Rhodnius prolixus in vitro. Journal of Insect Physiology 43: 651-656.
Van Buul-Offers, S.C. and Kooijman, R. 1998. The role of growth hormone and insulin-like growth factors in the
immune system. Cellular and Molecular Life Science 54, 1083-1094.
Vandesande, F, Dierickx, K. 1976. Immunocytochemical demostration of separate vasotocinergic neurons in the
amphibian magnocellular neurosecretory system. Cell and Tissue Research 179, 289-296.
Van de Velde, S., Badisco, L., Claeys, I., Verleyen, P., Chen, X., Vanden Bosch, L., Vanden Broeck, J., Smagghe,
G. 2007. Insulin-like peptides in Spodoptera littoralis (Lepidoptera): Detection, localization and
identification. General and Comparative Endocrinology 153, 72-79.
Vargas-Albores, F., Yepiz-Plascencia, G. Shrimp immunity. 1998. Trends in Comparative and Biochemical
Physiology 5, 195-210.
Weiss, I.M., Göhring, W., Fritz, M., Mann, K. 2001. Perlustrin, a Haliotis laevigata (abalone) nacre protein, is
homologous to the insulin-like growth factor binding protein N-terminal module of vertebrates.
Biochemical and Biophysical Research Communication 285, 244-249.
Wu, Q., Brown, M. R., 2006. Signaling and functions of Insulin-like peptides in insects. Annual Review of
Entomology 51, 1-24.
Zitnan, D., Sehnal, F., Mizoguchi, A., Ishizaki, H., Nagasawa, H., Suzuki, A. 1990. Developmental changes in the
bombyxin- and insulin-like immunoreactive neurosecretory system in the wax moth, Galleria mellonella.
Development Growth and Differentiation 32, 637-645.
