Summary
The axon terminals of the neuroendocrine caudo-dorsal cells (CDC) of the freshwater snail Lymnaea stagnalis release an ovulationstimulating hormone by exocytosis in a calcium-dependent way. Ultrastructural studies of the terminals, involving the K-pyroantimonate method for the demonstration of calcium and control tests with EGTA, show that calcium occurs in mitochondria and in various types of vesicular structure. Quantitative investigations indicate that mitochondria accumulate calcium during a short period of high neurohormone-release activity (“active state”; ca. 45 min) and release it again into the axoplasm some hours later, during a period of low secretory activity (“resting state”). Probably, in this way mitochondria play an important role in the buffering of the axoplasmic calcium concentration during high hormone-release activity. HRP-incorporation studies strongly suggest that the calcium-containing vesicular structures constitute a mechanism of membrane sequestration by which the CDC axon terminals resorb, transform, and release parts of the axolemma after exocytotic hormone release. The results furthermore indicate that this mechanism also may be involved in the control of the calcium concentration of the axoplasm, by taking up calcium from the axoplasm and releasing it into the extracellular space.
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References
Aguas AP, Nickerson PA (1981) Distribution of calcium differs in relaxed and contracted myocardial cells of the rat. Cell Tissue Res 221:295–302
Baker PF, McNaughton PA (1976) Kinetics and energetics of calcium efflux from intact squid giant axons. J Physiol 259:103–144
Bliss CI (1967) Statistics in biology, vol I. McGraw-Hill, New York
Borle AB (1973) Calcium metabolism at the cellular level. Fed Proc 32:1944–1950
Douglas WW (1974) Mechanisms of release of neurohypophyseal hormones: stimulus-secretion coupling. In: Greep RO, Astwood EB (eds) Handbook of physiology, sect 8, endocrinology, vol 4, part I. Williams and Wilkins, Baltimore, pp 191–224
Douglas WW, Nagasawa J, Schulz R (1971) Electron microscopic studies on the mechanism of secretion of posterior pituitary hormones and significance of microvesicles (“synaptic vesicles”): evidence of secretion by exocytosis and formation of microvesicles as a byproduct of this process. Mem Soc Endocrinol 19:353–378
Duce IR, Keen P (1978) Can neuronal smooth endoplasmic reticulum function as a calcium reservoir? Neuroscience 3:837–848
Geraerts WPM, Bohlken S (1976) The control of ovulation in the hermaphroditic freshwater snail Lymnaea stagnalis by the neurohormone of the Caudo-Dorsal Cells. Gen Comp Endocrinol 28:350–357
Geraerts WPM, Buma P, Hogenes TM (in press) Isolation of neurosecretory granules from the neurohaemal areas of peptidergic systems of Lymnaea stagnalis, with special reference to the ovulation hormone producing Caudo-Dorsal Cells. Gen Comp Endocrinol
Geuze JJ Kramer MF (1974) Function of coated membranes and multivesicular bodies during membrane regulation in stimulated exocrine pancreas. Cell Tissue Res 156:1–20
Graham RJ, Karnovsky MJ (1966) The early stage of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem 14:291–302
Greenawalt JW, Rossi CS, Lehninger AL (1964) Effect of active accumulation of calcium and phosphate ions on the structure of rat liver mitochondria. J Cell Biol 23:21–38
Heuser JE, Reese TS (1973) Evidence for recycling of synaptic vesicle membrane during transmitter release at the frog neuromuscular junction. J Cell Biol 57:315–344
Holtzman E, Peterson ER (1969) Uptake of protein by mammalian neurons. J Cell Biol 40:863–869
Holtzman E, Teichberg S, Abrahams SJ, Citkowitz E, Crain SM, Kawai N, Peterson ER (1973) Notes on synaptic vesicles and related structures, endoplasmic reticulum, lysosomes and peroxisomes in nervous tissue and the adrenal medulla. J Histochem Cytochem 21:349–385
Kits KS (1981) Electrical activity and hormone output of ovulation hormone producing neuroendochine cells in Lymnaea stagnalis (Gastropoda). In: Salanki J (ed) Neurobiology of invertebiates — mechanisms of integration. Adv Physiol Sci Vol 23. Akadémiai Kiadó, Budapest, pp 35–54
Kits KS, Maat A ter (1983) Neurophysiology of the peptidergic Caudo-Dorsal Cells in Lymnaea stagnalis. In: Lever J, Boer HH (eds) Molluscan neuro-endocrinology. Mon Royal Neth Academy of Arts and Sciences. North-Holland Publishing Company, Amsterdam Oxford New York, pp 60–68
Klein RL, Yen S-S, Thureson-Klein Å (1972) Critique on the K-pyroantimonate method for semiquantitive estimation of cations in conjunction with electron microscopy. J Histochem Cytochem 20:65–78
Komnick H (1962) Elektronenmikroskopische Lokalisation von Na+ und Cl- in Zellen und Geweben. Protoplasma 55:414–418
Lundquist I, Josefsson J-O (1971) Sensitive method for determination of peroxidase activity in tissue by means of coupled oxidation reaction. Anal Biochem 41:567–577
Morris JF, Nordmann JJ (1980) Membrane recapture after hormone release from nerve endings in the neural lobe of the rat pituitary gland. Neuroscience 5:639–649
Nagasawa J (1977) Exocytosis: the common release mechanism of secretory granules in glandular cells, neurosecretory cells, neurones and paraneurones. Arch Histol Jpn 40 (suppl.): 31–47
Nagasawa J, Douglas WW, Schulz RA (1971) Micropinocytotic origin of coated and smooth microvesicles (“synaptic vesicles”) in neurosecretory terminals of posterior pituitary glands demonstrated by incorporation of horseradish peroxidase. Nature 232:341–342
Nordmann JJ, Chevalier J (1980) The role of microvesicles in buffering (Ca2+)i in the neurohypophysis. Nature 287:54–56
Nordmann JJ, Morris JF (1976) Membrane retrieval at neurosecretory axon endings. Nature 261:723–725
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
Normann TC (1976) Neurosecretion by exocytosis. Int Rev Cytol 46:1–77
Normann TC, Hall TA (1978) Calcium and sulphur in neurosecretory granules and calcium in mitochondria as determined by electron microscope X-ray microanalysis. Cell Tissue Res 186:453–463
Normann TC, Samaranayaka-Ramasamy M (1977) Secretory hyperactivity and mitochondrial changes in neurosecretory cells of an insect. Cell Tissue Res 183:61–69
Pelletier G (1973) Secretion and uptake of peroxidase by rat adenohypophysial cells. J Ultrastruct Res 43:445–459
Roubos EW (1975) Regulation of neurosecretory activity in the freshwater pulmonate Lymnaea stagnalis (L.) with particular reference to the role of the eyes. Cell Tissue Res 160:291–314
Roubos EW (1976) Neuronal and non-neuronal control of the neurosecretory Caudo-Dorsal Cells of the freshwater snail Lymnaea stagnalis (L.). Cell Tissue Res 168:11–31
Roubos EW, Buma P (1982) Cellular mechanisms of neurohormone release in the snail Lymnaea stagnalis. In: Buijs RM, Pévet P, Swaab DF (eds) Progr Brain Res vol 55, Chemical transmission in the brain. Elsevier Biomedical Press, Amsterdam New York, pp 185–194
Roubos EW, Wal-Divendal RM van der (1980) Ultrastructural analysis of peptide-hormone release by exocytosis. Cell Tissue Res 207:267–275
Roubos EW, Boer HH, Schot LPC (1981 a) Peptidergic neurons and the control of neuroendocrine activity in the freshwater snail Lymnaea stagnalis (L.). In: Farner DS, Lederis K (eds) Neurosecretion ↣olecules, cells, systems. Plenum Press, New York London, pp 119–127
Roubos EW, Schmidt ED, Moorer-van Delft CM (1981 b) Ultrastructural dynamis of exocytosis in the ovulation-neurohormone producing Caudo-Dorsal Cells of the freshwater snail Lymnaea stagnalis (L.). Cell Tissue Res 215:63–73
Shapiro SS, Wilk MB (1965) An analysis of variance test for normality. Biometrika 52:591–611
Shaw FD, Morris JF (1980) Calcium localization in the rat neurohypophysis. Nature 287:56–58
Steel RGD, Torrie JH (1960) Principles and procedures of statistics. McGraw-Hill, New York Toronto London
Stoeckel ME, Hindelang-Gertner C, Dellmann H-D, Porte A, Stutinsky F (1975) Subcellular localization of calcium in the mouse hypophysis. I. Calcium distribution in the adenoand neurohypophysis under normal conditions. Cell Tissue Res 157:307–322
Vasington FD, Murphy JV (1962) Ca++ uptake by rat kidney mitochondria and its dependence on respiration and phosphorylation. J Biol Chem 237:2670–2677
Weakley BS (1979) A variant of the pyroantimonate technique suitable for localization of calcium in ovarian tissue. J Histochem Cytochem 27:1017–1028
Wendelaar Bonga SE (1970) Ultrastructure and histochemistry of neurosecretory cells and neurohaemal areas in the pond snail Lymnaea stagnalis (L.). Z Zellforsch 108:190–224
Wendelaar Bonga SE (1971) Formation, storage and release of neurosecretory material studied by quantitative electron microscopy in the freshwater snail Lymnaea stagnalis (L.). Z Zellforsch 113:490–517
With ND de (1977) Evidence for the independent regulation of specific ions in the haemolymph of Lymnaea stagnalis (L.). Proc Kon Ned Akad Wet C80:144–157
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This research was made possible by a grant from the Netherlands Organization for the Advancement of Pure Research (Z.W.O.)
The authors wish to thank Prof. H.H. Boer for stimulating interest during the study and helpful comments during the preparation of the manuscript, and Prof. J. Lever for critically reading the manuscript
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Buma, P., Roubos, E.W. Calcium dynamics, exocytosis, and membrane turnover in the ovulation hormone-releasing caudo-dorsal cells of Lymnaea stagnalis . Cell Tissue Res. 233, 143–159 (1983). https://doi.org/10.1007/BF00222239
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DOI: https://doi.org/10.1007/BF00222239