Summary
Sections of neurosecretory cells fixed in glutaraldehyde and osmium tetroxide were studied by means of an EMMA-4 analytical microscope. Secretory granules in neurosecretory cells of the corpus cardiacum and of the brain, both in the desert locust Schistocerca and in the blowfly Calliphora, as well as neurosecretory granules in posterior pituitaries of the frog Rana and of the albino rat all contain a high concentration of calcium. A distinct sulphur peak was also a constant feature.
In neurosecretory cells of the corpus cardiacum of Schistocerca the chromatin contained a high concentration of calcium. The mitochondria also contained much calcium, but part of this disappeared during preparation except when fixative and wash contained calcium chloride. By block staining with uranyl acetate most calcium is displaced from the mitochondria, whereas most of the calcium remains in the neurosecretory granules. Since the calcium peaks in spectra from neurosecretory granules appear of similar size, regardless of variations in the preparative procedure, this calcium must be firmly bound. The possible role of the calcium bound to the neurosecretory substance is discussed.
The presence of sulphur in insect neurosecretory granules indicates the presence of a protein besides the hormone, i.e., an insect neurophysin.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Adams, C.W.M., Sloper, J.C.: Technique for demonstrating neurosecretory material in the human hypothalamus. Lancet 1955I, 651–652
Arvy, L., Gabe, M.: Histochemistry of the neurosecretory product of the pars intercerebralis of pterygote insects. In: Neurosecretion (H. Heller and R.B. Clark, eds.), pp. 331–344. New York: Academic Press 1962
Baker, P.F.: Excitation-secretion coupling. Recent Advan. Physiol. 9, 51–86 (1974)
Bloch, R.J., Van Dyke, H.B.: Amino acids in posterior pituitary protein. Arch. Biochem. Biophys. 36, 1–4 (1952)
Borle, A.B.: Calcium metabolism at the cellular level. Fed. Proc. 32, 1944–1950 (1973)
Borowitz, J.L.: Effect of acetylcholine on the subcellular distribution of 45Ca in bovine adrenal medulla. Biochem. Pharmacol. 18, 715–723 (1969)
Borowitz, J.L., Fuwa, K., Weiner, N.: Distribution of metals and catecholamines in bovine adrenal medulla subcellular fractions. Nature (Lond.) 205, 42–43 (1965)
Carafoli, E., Lehninger, A.L.: A survey on the interaction of calcium ions with mitochondria from different tissues and species. Biochem. J. 122, 681–690 (1971)
Du Vigneaud, V., Lawler, H.C., Popenoe, E.A.: Enzymatic cleavage of glycinamide from vasopressin and a proposed structure for this pressor-antidiuretic hormone of the posterior pituitary. J. Amer. chem. Soc. 75, 4880–4881 (1953a)
Du Vigneaud, V., Ressler, C., Trippett, C.: The sequence of amino acids in oxytocin with a proposal for the structure of oxytocin. J. biol. Chem. 205, 949–957 (1953b)
Hall, T.A., Anderson, H.C., Appleton, T.C.: The use of thin specimens for X-ray microanalysis in biology. J. Microscopy 99, 177–182 (1973)
Helle, K.B., Serck-Hanssen, G.: Matrix phase organization at the molecular level in the chromaffin granule. Abstr. 3. Int. Catecholamine Symposium, Strassbourg (1973)
Johnson, R.G., Scarpa, A.: Ion permeability of isolated chromaffin granules. J. gen. Physiol. 68, 601–631 (1976)
Krogh, I.M., Normann, T.C.: The corpus cardiacum neurosecretory cells of Schistocerca gregaria. Electron microscopy of resting and secreting cells. Acta zool. 58, 69–78 (1977)
Lazarewicz, J.W., Haljamäe, H., Hamberger, A.: Calcium metabolism in isolated brain cells and subcellular fractions. J. Neurochem. 22, 33–46 (1974)
Lust, W.D., Robinson, J.D.: Calcium accumulation by isolated nerve ending particles from brain. I. The site of energy-dependent accumulation. J. Neurobiol. 1, 303–316 (1970)
Normann, T.C.: Neurosecretion by exocytosis. Int. Rev. Cytol. 46, 1–77 (1976)
Normann, T.C., Samaranayaka-Ramasamy, M.: Secretory hyperactivity and mitochondrial changes in neurosecretory cells of an insect. Cellular effects of the insecticide lindane. Cell Tiss. Res., 183, 61–69 (1977)
Oschman, J.L., Hall, T.A. Peters, P.D., Wall B.J.: Association of calcium with membranes of squid giant axon. J. Cell Biol. 61, 156–165 (1974)
Prentø, P.: Histochemistry of neurosecretion in the pars intercerebralis-corpus cardiacum system of the desert locust Schistocerca gregaria. Gen. comp. Endocr. 18, 482–500 (1972)
Russell, J.T., Thorn, N.A.: Adenosine triphosphate dependent calcium uptake by subcellular fractions from bovine neurohypophyses. Acta physiol. scand 93, 364–377 (1975)
Russell, J.T., Thorn, N.A.: Isolation and purification of calcium binding proteins from bovine neurohypophyses. Biochim. biophys. Acta (Amst.) (in press)
Serck-Hanssen, G., Christiansen, E.: Uptake of calcium in chromaffin granules of bovine adrenal medulla stimulated in vitro. Biochim. biophys. Acta (Amst.) 307, 404–415 (1973)
Skaer, R.J., Peters, P.D., Emmines, J.P.: The localization of calcium and phosphorus in human platelets. J. Cell Sci. 15, 679–692 (1974)
Steel, C.G.H., Morris, G.P.: Evidence from X-ray microanalysis of sulphur-rich protein in insect neurosecretion: a new approach to the estimation of neurosecretory activity. Gen. comp. Endocr. 26, 517–524 (1975)
Stone, J.V., Mordue, W., Batley, K.E., Morris, H.R.: Structure of locust adipokinetic hormone, a neurohormone that regulates lipid utilisation during flight. Nature (Lond.) 263, 207–211 (1976)
Takaya, K.: Energy dispersive X-ray microanalysis of neurosecretory granules of mouse pituitary on fresh air-dried tissue spreads. Cell Tiss. Res. 159, 227–231 (1975a)
Takaya, K.: Electron probe microanalysis of the dense bodies of human blood platelets. Arch. histol. jap. 37, 335–341 (1975b)
Takaya, K.: Electron microscopy of unstained, fresh air-dried spreads of mouse pancreas acinar cells and energy dispersive X-ray microanalysis of zymogen granules. Cell Tiss. Res. 166, 117–123 (1976)
Thorn, N.A.: Role of calcium in secretory processes. In: Secretory mechanism of exocrine glands (N.A. Thorn and D.H. Petersen, eds.), pp. 305–330. Copenhagen: Munksgaard 1974
Thorn, N.A., Russell, J.T., Vilhardt, H.: Hexosamine, calcium, and neurophysin in secretory granules and the role of calcium in hormone release. Ann. N.Y. Acad. Sci. 248, 202–217 (1975)
Wallach, D., Schramm, M.: Calcium and the exportable protein in rat parotid gland. Parallel subcellular distribution and concomitant secretion. Europ. J. Biochem. 21, 433–437 (1971)
Yarom, R., Hall, T.A., Peters, P.D.: Calcium in myonuclei: Electron probe X-ray analysis. Experientia (Basel) 31, 154–157 (1975)
Author information
Authors and Affiliations
Additional information
We wish to thank Dennis Greer for the operation of the analytical microscope. This work was supported by a “Wellcome-Carlsberg Travelling Research Fellowship” awarded to T.N. The EMMA-4 facility is supported by a grant from the British Science Research Council
Rights and permissions
About this article
Cite this article
Christian Normann, T., Hall, T.A. Calcium and sulphur in neurosecretory granules and calcium in mitochondria as determined by electron microscope X-ray microanalysis. Cell Tissue Res. 186, 453–463 (1978). https://doi.org/10.1007/BF00224934
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00224934