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The ultrastructure of nerve endings containing pigment-dispersing hormone (PDH) in crustacean sinus glands: Identification by an antiserum against a synthetic PDH

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Summary

A high-liter antiserum has been obtained from two rabbits immunized with a glutaraldehyde conjugate of synthetic pigment-dispersing hormone (PDH) from Uca pugilator and bovine thyroglobulin. The antiserum blocked melanophore-dispersing activity of the peptide in vivo. In sinus glands (SG) of Carcinus maenas, Cancer pagurus, Uca pugilator and Orconectes limosus, electron-microscopic immunocytochemistry revealed sparsely distributed axon endings containing a distinct PDH-immunoreactive type of neurosecretory granules (diameter 90–130 nm). Exocytotic figures indicating release of the content of these granules into hemolymph lacunae were occasionally observed. Preservation of fine structure and antigenicity of the PDH granules were markedly dependent on the fixation procedure used. A preliminary experiment with C. maenas showed that preterminal axon dilatations near the basal lamina seemed to accumulate PDH-granules when animals were kept in complete darkness for three days. Immunodot blotting of fractions after high pressure liquid chromatography (HPLC) of extracts from SGs of C. maenas and O. limosus revealed a strongly immunoreactive substance at a retention time very similar to those of synthetic PDHs of Uca pugilator and Pandalus borealis. It is also coincident with a zone of biological activity. Thus, the antigen demonstrated by immunocytochemistry is identical or very similar to one of the known PDHs.

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References

  • Aoto I, Hisano S (1985) Ultrastructural evidence for the existence of the distal retinal pigment light-adapting hormone in the sinus gland of the prawn Palaemon paucidens. Gen Comp Endocrinol 60:468–475

    Google Scholar 

  • Bendayan M, Zollinger M (1983) Ultrastructural localization of antigenic sites on osmium-fixed tissues applying the protein A-gold technique. J Histochem Cytochem 31:101–109

    Google Scholar 

  • Bunt AH (1969) Formation of coated and “synaptic” vesicles within neurosecretory axon terminals of the crustacean sinus gland. J Ultrastr Res 28:411–421

    Google Scholar 

  • DeMey J, Moeremans M, Genens G, Nuyders R, De Brabander M (1981) High resolution light and electron microscopic localization of tubulin with the IGS (immuno gold staining) method. Cell Biol Int Rep 5:889–899

    Google Scholar 

  • Fernlund P (1971) Chromactivity hormones of Pandalus borealis. Isolation and purification of a light-adapting hormone. Biochem Biophys Acta 237:519–529

    Google Scholar 

  • Fernlund P (1976) Structure of a light-adapting hormone from the shrimp, Pandalus borealis. Biochem Biophys Acta 439:17–25

    Google Scholar 

  • Fernlund P, Josefsson L (1972) Crustacean color-change hormone: Amino acid sequence and chemical synthesis. Science 177:173–174

    Google Scholar 

  • Frens G (1973) Controlled nucleation for the regulation of the particle size in monodisperse gold solutions. Nature Phys Sci 241:20–22

    Google Scholar 

  • Gorgels-Kallen JL, Van Herp F (1981) Localization of crustacean hyperglycemic hormone (CHH) in the X-organ sinus gland complex in the eyestalk of the crayfish, Astacus leptodactylus. J Morphol 170:347–355

    Google Scholar 

  • Gorgels-Kallen JL, Van Herp F, Leuven RSEW (1982) A comparative immunocytochemical investigation of the crustacean hyperglycemic hormone (CHH) in the eyestalks of some decapod crustacea. J Morphol 174:161–169

    Google Scholar 

  • Greenberg MJ, Price DA (1983) Invertebrate neuropeptides: native and naturalized. Ann Rev Physiol 45:271–288

    Google Scholar 

  • Hearn SA, Silver MM, Sholdice JA (1985) Immunoelectron microscopic labelling of immunoglobulin in plasma cells after osmium fixation and epoxy embedding. J Histochem Cytochem 33:1212–1218

    Google Scholar 

  • Hogben L, Slome D (1931) The pigmentary effector system. VI. The dual character of endocrine co-ordination in amphibian colour change. Proc R Soc London Ser B 108:10–53

    Google Scholar 

  • Jaros PP, Keller R (1979) Immunocytochemical identification of hyperglycemic hormone-producing cells in the eyestalk of Carcinus maenas. Cell Tissue Res 204:379–385

    Google Scholar 

  • Jaros PP, Dircksen H, Keller R (1985) Occurrence of immunoreactive enkephalins in a neurohemal organ and other nervous structures in the eyestalk of the shore crab, Carcinus maenas L. (Crustacea, Decapoda). Cell Tissue Res 241:111–117

    Google Scholar 

  • Keller R (1977) Comparative electrophoretic studies of crustacean neurosecretory hyperglycemic and melanophore-stimulating hormones from isolated sinus glands. J Comp Physiol 122:359–373

    Google Scholar 

  • Keller R (1983) Biochemistry and specificity of the neurohemal hormones in Crustacea. In: Gupta AT (ed) Neurohemal organs of arthropods Charles C. Thomas, Springfield, Illinois pp 118–148

    Google Scholar 

  • Keller R, Kegel G (1984) Studies on crustacean neuropeptides by use of high performance liquid chromatography. In: Hoffmann JA, Porchet M (eds) Biosynthesis, metabolism and mode of action of invertebrate hormones. Springer, Berlin Heidelberg New York pp 145–154

    Google Scholar 

  • Keller R, Jaros PP, Kegel G (1985) Crustacean hyperglycemic neuropeptides. Amer Zool 25:207–221

    Google Scholar 

  • Kleinholz LH (1970) A progress report on the separation and purification of crustacean neurosecretory pigmentary effector hormones. Gen Comp Endocr 14:578–588

    Google Scholar 

  • Kleinholz LH (1972) Comparative studies of crustacean melanophore stimulating hormones. Gen Comp Endocr 19:473–483

    Google Scholar 

  • Kleinholz LH, Keller R (1979) Endocrine regulation in Crustacea. In: Barrington EJW (ed) Hormones and evolution. Academic Press NY pp 158–213

    Google Scholar 

  • Kleinholz LH, Rao KR, Riehm JP, Tarr GE, Johnson L, Norton S (1986) Isolation and sequence analysis of a pigment dispersing hormone from eyestalks of the crab, Cancer magister. Biol Bull 170:135–143

    Google Scholar 

  • Langford LA, Coggeshall RE (1980) The use of potassium ferricyanide in neural fixation. Anat Rec 197:297–303

    Google Scholar 

  • Larsson LI (1981) A novel immunocytochemical model system for specificity and sensitivity screening of antisera against multiple antigens. J Histochem Cytochem 29:408–410

    Google Scholar 

  • Mangerich S, Keller R, Dircksen H (1986) Immunocytochemical identification of structures containing putative red pigmentconcentrating hormone in two species of decapod crustaceans. Cell Tissue Res 245:377–386

    Google Scholar 

  • Mangerich S, Keller R, Dircksen H, Rao KR, Riehm JP (1987) Immunocytochemical localization of pigment-dispersing hormone (PDH) and its coexistence with FMRFamide-immunoreactive material in the eyestalks of the decapod crustaceans Carcinus maenas and Orconectes limosus. Cell Tissue Res 250:365–375

    Google Scholar 

  • Maxwell MH (1977) Two rapid and simple methods used for removal of resins from 1.0 μm thick epoxy sections. J Microsc (Oxford) 112:253–255

    Google Scholar 

  • Moeremans M, Daneels G, Van Dijck A, Langanger G, De Mey J (1984) Sensitive visualization of antigen-antibody reactions in dot and blot immune overlay assays with immunogold and immunogold silver staining. J Immunol Meth 74:353–360

    Google Scholar 

  • Nordmann JJ (1977) Ultrastructural appearance of neurosecretory granules in the sinus gland of the crab after different fixation procedures. Cell Tissue Res 185:557–563

    Google Scholar 

  • Nordmann JJ (1979) Hormone release and membrane retrieval in neurosecretion. In: Colloques Internationaux du CNRS No 280 Biol Cellulaire des processus neurosécrétaires hypothalamiques Chapitre 37 pp 614–636

  • Nordmann JJ, Morris JF (1976) Membrane retrieval in neurosecretory axon endings. Nature 261:723–725

    Google Scholar 

  • Nordmann JJ, Morris JF (1980) Depletion of neurosecretory granules and membrane retrieval in the sinus gland of the crab. Cell Tissue Res 205:31–42

    Google Scholar 

  • Nordmann JJ, Labouesse J (1981) Neurosecretory granules: Evidence of an aging process within the neurohypophysis. Science 211:595–597

    Google Scholar 

  • Pérez-González MD (1957) Evidence for hormone-containing granules in sinus glands of the fiddler crab Uca pugilator. Biol Bull 113:426–441

    Google Scholar 

  • Rao KR (1985) Pigmentary effectors. In: Bliss DE, Mantel LH (eds) The Biology of Crustacea vol 9, Academic Press pp 395–462

  • Rao KR, Riehm JP, Zahnow CA, Kleinholz LH, Tarr GE, Johnson L, Norton S, Landau M, Semmes OJ, Sattelberg RM, Jorenby WH, Hintz MF (1985) Characterization of a pigment-dispersing hormone in eyestalks of the fiddler crab Uca pugilator. Proc Nat Sci USA 82:5319–5323

    Google Scholar 

  • Smith G (1974) The ultrastructure of the sinus gland of Carcinus maenas (Crustacea: Decapoda). Cell Tissue Res 155:117–125

    Google Scholar 

  • Spurr AR (1969) A low-viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res 26:31–43

    Google Scholar 

  • Sternberger LA (1974) Immunocytochemistry. Prentice Hall Inc Englewood Cliffs, NJ

    Google Scholar 

  • Unglaub-Silverthorn S (1975) Neurosecretion in the sinus gland of the fiddler crab, Uca pugilator. Cell Tissue Res 165:129–133

    Google Scholar 

  • Van Harreveld A (1936) A physiological solution for freshwater crustacean. Proc Soc Exp Biol Ned 34:428–432

    Google Scholar 

  • Van Herp F, Van Buggenum HJM (1979) Immunocytochemical localization of hyperglycemic hormone (HGH) in the neurosecretory system of the eyestalk of the crayfish Astacus leptodactylus. Experientia 35:1527–1529

    Google Scholar 

  • Webster SG, Keller R (1987) Physiology and biochemistry of crustacean neurohormonal peptides. In: Thorndyke M, Goldsworthy G (eds) Amines and peptides in invertebrates, Cambridge University Press (in press)

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Authors and Affiliations

  1. Rheinische Friedrich-Wilhelms-Universität, Institut für Zoophysiologie, Bonn, Federal Republic of Germany

    Heinrich Dircksen, Gabriele Gaus & Rainer Keller

  2. Department of Biology, The University of West Florida, Pensacola, Florida, USA

    Cynthia A. Zahnow, K. Ranga Rao & John P. Riehm

Authors
  1. Heinrich Dircksen
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  2. Cynthia A. Zahnow
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  3. Gabriele Gaus
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  4. Rainer Keller
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  5. K. Ranga Rao
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  6. John P. Riehm
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Dircksen, H., Zahnow, C.A., Gaus, G. et al. The ultrastructure of nerve endings containing pigment-dispersing hormone (PDH) in crustacean sinus glands: Identification by an antiserum against a synthetic PDH. Cell Tissue Res. 250, 377–387 (1987). https://doi.org/10.1007/BF00219082

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