Sea urchin egg cortices were used as an in vitro natural membrane model system to determine the effects of inhalation anesthetics on the Ca2+-regulated exocytotic fusion of cortical vesicles with the egg plasma membrane. When Ca2+ was either absent or present in amounts below the threshold for exocytosis, methoxyflurane, halothane, enflurane, isoflurane, chloroform and fluoroxene, at concentrations up to S mM, had no effect on the fusion of cortical vesicles with the plasma membrane. However, when Ca2+ was present at or above threshold levels for exocytosis, each of the tested anesthetics caused an inhibition of cortical vesicle fusion. Exocytosis was inhibited most effectively by methoxyflurane (55%), followed by halothane (30%), while fuoroxene consistently had the least effect (< 5%). These observations support the view that volatile anesthetics can impair the Ca2+-regulated fusogenic activities of natural membranes and are consistent with other data showing that inhalational agents inhibit secretory processes in intact cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- PIPES:
-
piperazine-N-N′-bis (2-ethane sulfonic acid)
- PMSF:
-
phenylmethylsulfonylfluoride
- SW:
-
sea water
- TAPS:
-
trishydroxymethyl-methylaminopropane sulfonic acid
References
BEELER, T. and GABLE, K. (1985). Effect of halothane on Ca2+-induced Ca2+ release from sarcoplasmic reticulum vesicles isolated from rat skeletal muscle. Biochim. Biophys. Acta 821:142–152.
BOSNJAK, Z.J., SEAGARD, J.L., WU, A. and KAMPINE, J.P. (1982). The effects of halothane on sympathetic ganglionic transmission. Anesthesiology 57:473–479.
CHEEK, D.B.C., KETTELKAMP, N.S., AUSTIN, D.R., DOWNES, H. and HIRSHMAN, C.A. (1986). Inhibition of d-tubocurarine induced histamine release by halothane. Anesthesiology 65:A334.
DECKER, S.J. and KKINSEY, W.H. (1983). Characterization of cortical secretory vesicles from the sea urchin egg. Dev. Biol. 96:37–45.
DeLISLE, R.C. and WILLIAMS, J.A. (1986). Regulation of membrane Fusion in secretory exocytosis. Ann. Rev. Physiol. 48:225–238.
DLUZEWSKI, A.R., HALSEY, M.J. and SIMMONDS, A.C. (1983). Membrane interactions with general and local anesthetics: A review of molecular hypotheses of anesthesia. Mol. Aspects Med. 6:459–573.
EPEL, D. and VACQUIER, V.D. (1978). Membrane fusion events during invertebrate fertilization. In: Cell Surface Reviews: Membrane Fusion (G. Poste and G. Nicholson, eds.), Vol 5, pp163. Elsevier/North Holland Biomedical Press, Amsterdam.
FIRESTONE, L.L. and KITZ, R.J. (1986). Anesthetics and lipids: some molecular perspectives. Seminars in Anesthesia 4:286–300.
GOTHERT, M., DORN, W. and LOEWENSTEIN, I. (1976). Inhibition of catecholamine release from the adrenal medulla by halothane. Site and mechanism of action. NaunynSchmiedebergs Arch. Pharmacol. 294:239–249.
HAGGERTY, J.C. and JACKSON, R.C. (1983). Release of granule contents from seaurchin egg cortices. New assay procedures and inhibition by sulfhydryl-modifying agents. J. Biol. Chem. 258:1819–1825.
HALSEY, M.J. (1980). Physicochemical properties of inhalational anesthetics. In: General Anesthesia (Gray, T.C., Nunn, J.F. and Utting, J.E., eds.), Vol. 1, pp. 45–65. Butterworth Inc., Boston.
HINKLEY, R.E. and CHAMBERS, E.L. (1982). Structural changes in dividing sea urchin eggs induced by the volatile anesthetic halothane. J. Cell Sci. 55:327–39.
HINKLEY, R.E. and WRIGHT, B.D. (1985). Comparative effects of halothane, enflurane, and methoxyflurane on the incidence of abnormal development using sea urchin gametes as an in vitro model system. Anesth. Analg. 64:1005–1009.
HINKLEY, R.E. and WRIGHT, B.D. (1986). Effects of the volatile anesthetic halothane on fertilization and early development in the sea urchin Lytechinus variegatus: Evidence that abnormal development is due to polyspermy. Teratology 34:291–301.
HUNT, G.R.A. and JONES, I.C. (1983). A H-NMR investigation of the effects of ethanol and general anesthetics on ion channels and membrane fusion using unilamellar phospholipid membranes. Biochim. Biophys. Acta 736:1–10.
LERMAN, J., WILLIS, M.M., GREGORY, G.A. and EGER, E.I. (1983). Osmolarity determines the solubility of anesthetics in aqueous solutions at 37°C. Anesthesiology 59:554–558.
LOWRY, O.H., ROSEBROUGH, N.J., FARR, A.L. and RANDALL, R.J. (1951). Protein measurement with the folin phenol reagent. J. Biol. Chem. 193:265–275.
MAZIA, D., SCHATTEN, G. and SALE, W. (1975). Adhesion of cells to surfaces coated with poly-l-lysine. Applications to electron microscopy. J. Cell Biol. 66:198–200.
MEIJER, L., DUBE, F. and GUERRIER, P. (1983). Trifluoperazine-sensitive step during sea urchin, echiuroid and pelcypod egg activation. Develop. Growth and Differ. 25:469–475.
OGAWA, Y. and KUREBAYASHI, N. (1982). The calcium-releasing action of halothane on fragmented sarcoplasmic reticulum. J. Biochem. 92:899–905.
POCOCK, G. and RICHARDS, C.D. (1986). Halothane inhibits catecholamine secretion and ion movements in bovine adrenal medullary cells. J. Physiol Lond. 384:25.
ROTH, S.H. and MILLER, K.W. (1986). Molecular and cellular mechanisms of anesthetics, pp. 459–470. Plenum, New York.
ROBERTSON, S. and POTTER, J.D. (1984). The regulation of free Ca2+ ion concentration by metal chelators. In: Methods in Pharmacology (A. Schwartz, ed.), Vol 5, pp. 63–75. Plenum, New York.
SASAKI, H. (1984). Modulation of calcium sensitivity by a specific cortical protein during sea urchin egg cortical vesicle exocytosis. Dev. Biol. 101:125–135.
SASAKI, H. and EPEL, D. (1983). Cortical vesicle exocytosis in isolated cortices of sea urchin eggs: description of a turbidometric assay and its utilization in studying effects of different media on discharge. Dev. Biol. 98:327–337.
SCHUELL, H. (1984). The prevention of polyspermic fertilization in sea urchins. Biol. Bull 167:271–309.
SIMMONDS, A.C. and HALSEY, M.J. (1986). General and local anesthetics perturb the fusion of phospholipid vesicles. Biochim. Biophys. Acta 813:331–337.
STEINHARDT, R.A. and ALDERTON, J.M. (1982). Calmodulin confers calcium sensitivity on secretory exocytosis. Nature, Lond. 295:154–155.
SUMIKAWA, K., HIRANO, H., AMAKATA, Y., AMENOMORI, Y. and AMAKATA, Y. (1985). Mechanism of the differential effects of halothane on nicotinic- and muscarinic-receptor-mediated response of the dog adrenal medulla. Anesthesiology 57:444–450.
UEDA, I., HIRAKAWA, M., ARAKAWA, K. and KAMAYA, H. (1986). Do anesthetics fluidize membranes? Anesthesiology 64:67–71.
VACQUIER, V.D. (1975). The isolation of intact cortical granules from sea urchin eggs: calcium ions trigger granule discharge. Dev. Biol. 43:62–74.
WHITAKER, M.J. and BAKER, P.F. (1983). Calcium-dependent exocytosis in an in vitro secretory granule plasma membrane preparation from sea urchin eggs and the effects of some inhibitors of cytoskeletal function. Proc. R. Soc. Lond. B. 218:397–413.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Lederhaas, G., Hinkley, R.E. The effects of inhalation anesthetics on calcium-stimulated exocytosis in a natural membrane model system. Cell Biol Toxicol 4, 149–161 (1988). https://doi.org/10.1007/BF00119242
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00119242