Summary
An attempt was made to study the subcellular localization of calcium in carotid body glomus cells of adult rats using fixation with glutaraldehyde followed by treatment with a mixture of pyroantimonate and osmium tetroxide. Precipitates were seen as electron-dense particles (EDP) in the glomus cells, mostly within membrane-bound organelles, such as dense-cored vesicles, mitochondria, small clear vesicles, multivesicular bodies, and especially in lysosomes. However, EDP were also seen in the nuclei and in the free cytoplasm of the glomus cells and even outside them.
Preincubation of carotid bodies in media containing calcium and either high potassium or calcium-ionophore A 23187 resulted in a marked increase in the general precipitation pattern, there being an increased amount of EDP both in the glomus cell nuclei and in the cytoplasm. Dense-cored vesicles more often showed precipitates than those in the controls. Some dense-cored vesicles contained multiple precipitates, typically located in the electron-lucent area between core and vesicle membrane.
Extensive diffusion of ions probably occurred during fixation before precipitation, making the localization of calcium and other precipitating cations unreliable. However, it is possible that precipitates, which were regularly seen in the dense-cored vesicles, may reflect the content of bound calcium. The possible significance of calcium in glomus cell function is discussed, and the need for more adequate methods is emphasized.
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References
Chan SY, Ochs SR, Jersild R Jr (1979) Calcium localization in mammalian nerve fibers in relation to its regulation and axoplasmic transport. Soc Neurosci Abs 5:59
Cramer EB, Cardasis C, Pereira G, Milks L, Ford D (1978) Ultrastructural localization of cations in the rat pars distalis under various experimental conditions. Neuroendocrinology 26:72–84
Davis WL, Matthews JL, Martin JH (1974) An electron microscopic study of myofilament calcium binding sites in native, EGTA-chelated and calcium reloaded glycerolated mammalian skeletal muscle. Calcif Tissue Res 14:139–152
Davis WL, Jones RG, Hagler HK (1979) Calcium containing lysosomes in the normal chick duodenum: A histochemical and analytical electron microscopic study. Tissue Cell 11:127–138
Duce IR, Keen P (1978) Can neuronal smooth endoplasmic reticulum function as a calcium reservoir? Neuroscience 3:837–848
Ellisman MH, Friedman PL, Hamilton WJ (1980) The localization of sodium and calcium to Schwann cell paranodal loops at nodes of Ranvier and of calcium to compact myelin. J Neurocytol 9:185–205
Fujita T (1977) Concept of paraneurons. In: Kobayashi S, Chiba T (eds) Paraneurons. New concepts on neuroendocrine relatives. Arch Histol Jpn 40:Suppl, pp 1–12
Grönblad M, Åkerman KE, Eränkö O (1980a) Ultrastructural evidence of exocytosis from glomus cells after incubation of adult rat carotid bodies in potassium-rich calcium-containing media. Brain Res 189:576–581
Grönblad M, Åkerman KE, Eränkö O (1980b) Exocytosis of amine-storing granules from glomus cells of the rat carotid body induced by incubation in K-rich media or in media containing Ca and ionophore A 23187. In: Eränkö O, Soinila S, Päivärinta H (eds) Histochemistry and Cell Biology of Autonomie Neurons, SIF Cells and Paraneurons. Raven Press, New York, pp 227–235
Hansen JT, Smith NKR (1979) Calcium binding sites in the vesicles of the carotid body and aortic body chief cells. Cell Tissue Res 199:145–151
Hayat MA (1975) Principles and techniques of scanning electron microscopy, Vol 4. Van Nostrand Reinhold Company, New York
Hellman B, Andersson T, Berggren P-O, Flatt P, Gylfe E, Kohnert K-D (1979) The role of calcium in insulin secretion. In: Dumont J, Nunez J (eds) Hormones and Cell Regulation, Vol 3. Elsevier/North Holland Biomedical Press, pp 69–96
Hess A (1977) The calcium binding sites of dense-core vesicles in the catecholaminergic glomus cells of the rat carotid body. Brain Res 138:555–560
Kanno T (1977) Physiology of paraneurons. In: Kobayashi S, Chiba T (eds) Paraneurons. New concepts on neuroendocrine relatives. Arch Histol Jpn 40:Suppl, pp 13–29
Klein RL, Yen S-S, Thureson-Klein Å (1972) Critique on the K-pyroantimonate method for semiquantitative estimation of cations in conjunction with electron microscopy. J Histochem Cytochem 20:65–78
Kohnert K-D, Hahn H-J, Gylfe E, Borg H, Hellman B (1979) Calcium and pancreatic B-cell function. 6. Glucose and intracellular 45Ca distribution. Mol Cell Endocrinol 16:205–220
Kostron H, Winkler H, Geissler D, König P (1977) Uptake of calcium by chromaffin granules in vitro. J Neurochem 28:487–493
Krammer EB (1978) Carotid body chemoreceptor function: Hypothesis based on a new circuit model. Proc Natl Acad Sci USA 75:2507–2511
McDonald DM, Mitchell RA (1975) The innervation of glomus cells, ganglion cells and blood vessels in the rat carotid body: a quantitative ultrastructural analysis. J Neurocytol 4:177–230
Nagasawa J (1977) Exocytosis: The common release mechanism of secretory granules in glandular cells, neurosecretory cells, neurons and paraneurons. In: Kobayashi S, Chiba T (eds) Paraneurons. New concepts on neuroendocrine relatives. Arch Histol Jpn 40:Suppl, pp 31–47
Oschman JL, Wall BJ (1972) Calcium-binding to intestine membranes. J Cell Biol 55:58–73
Pfeiffer DR, Taylor RW, Lardy HA (1978) Ionophore A 23187: Cation binding and transport properties. Ann NY Acad Sci 307:402–423
Serck-Hansen G, Christiansen EN (1973) Uptake of calcium in chromaffin granules of bovine adrenal medulla stimulated in vitro. Biochem Biophys Acta 307:404–414
Simson JAV, Spicer SS (1975) Selective subcellular localization of cations with variants of the potassium (pyro)antimonate technique. J Histochem Cytochem 23:575–598
Smith AD, Winkler H (1972) Fundamental mechanisms in the release of catecholamines. In: Blaschko H, Muscholl E (eds) Catecholamines. Springer-Verlag, Berlin Heidelberg New York, pp 538–617
Tóth I (1978) Comparative evaluation of different techniques used to detect the elemental composition of nervous structures by X-ray microanalysis. Proc Roy Microsc Soc, Micro 78:Suppl, 33 (Abstr)
Van Iren F, Van Essen-Joolen L, Van der Duyn Schouten P, Boers-Van der Sluijs P, de Bruijn WC (1979) Sodium and calcium localization in cells and tissues by precipitation with antimonate: A quantitative study. Histochemistry 63:273–294
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The present study has been supported by grants from the Finska Läkaresällskapet and the Sigrid Jusélius Foundation, Helsinki, Finland
We wish to express our gratitude to Dr. Robert Hamill of Eli Lilly Co. for kindly providing us with the ionophore A 23187. Technical assistance by Mrs. S. Huhtaniitty and Mrs. T. Stjernvall is gratefully acknowledged
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Grönblad, M., Åkerman, K.E.O. & Eränkö, O. Electron-dense precipitates in glomus cells of rat carotid body after fixation in glutaraldehyde and pyroantimonate-osmium tetroxide mixture as possible indicators of calcium localization. Cell Tissue Res. 217, 93–104 (1981). https://doi.org/10.1007/BF00233829
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DOI: https://doi.org/10.1007/BF00233829