Skip to main content
SpringerLink
Log in

Amiodarone and desethylamiodarone increase intrasynaptosomal free calcium through receptor mediated channel

  • Published:
Naunyn-Schmiedeberg's Archives of Pharmacology Aims and scope Submit manuscript

Summary

Long term amiodarone (AM) therapy has been associated with several side effects including neurotoxicity. Since AM alters Ca2+ regulated events, we have studied its effects on the compartmentation of free Ca2+ in the synaptosomes as an attempt to understand the mechanism of AM and its metabolite, desethylamiodarone (DEA)-induced neurotoxicity. Intact brain synaptosomes were prepared from male Sprague-Dawley rats. Both AM and DEA produced a concentration dependent increase in intrasynaptosomal free Ca2+ concentration ([Ca2+]i) to micromolar levels. The increase in [Ca2+]i was not transient and a steady rise was observed with time. Omission of Ca2+ from the external medium prevented the AM- and DEA-induced rise in [Ca2+]i suggesting that AM and DEA increased the intracellular [Ca2+]i due to increased influx of Ca2+ from external medium. AM- and DEA-induced increase in intrasynaptosomal [Ca2+]i was neither inhibited by a calcium channel blocker, verapamil, nor with a Na+ channel blocker, tetrodotoxin. However, the blockade of [Ca2+]i rise by AM and DEA was observed with MK-801, a receptor antagonist indicating that AM and DEA induced rise in [Ca2+]i is through receptor mediated channel. Both AM and DEA also inhibited N-methyl-D-aspartic acid (NMDA)-receptor binding in synaptic membranes in a concentration dependent manner, DEA being more effective, indicating that AM and DEA compete for the same site as that of NMDA and confirm the observation that these drugs increase intrasynaptosomal [Ca2+]i through receptor mediated channel. 45Ca accumulation into brain microsomes and mitochondria was significantly inhibited by AM and DEA, but without any effect on the Ca2+ release from these intracellular organelles. Also, both these drugs did not interfere with inositol 1,4,5-trisphosphate induced Ca2+ release from microsomes even at 10 μM concentration. These results clearly indicate that both AM and DEA increase intrasynaptosomal [Ca2+]i by an action on receptor mediated channel in plasma membrane, but not due to the release of Ca2+ from intracellular storage sites. This initial rise in [Ca2+]i, together with other changes in Ca2+ homeostasis, might be responsible for AM and DEA-induced neurotoxicity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Bagchi N, Brown TR, Schneider DS, Benerjee SK (1987) Effect of amiodarone on rat heart myosin isoenzymes. Circ Res 60: 621–625

    Google Scholar 

  • Berridge MJ (1987) Inositol lipids and cell proliferation. Biochim Biophys Acta 907: 33–45

    Google Scholar 

  • Berthon B, Binet A, Mauger JP, Claret M (1984) Cytosolic free calcium in isolated rat hepatocytes as measured by quin-2: effects of noradrenaline and vasopressin. FEBS Lett 167: 19–24

    Google Scholar 

  • Bondy SC (1989) Intracellular calcium and neurotoxic events. Neurotoxicol Teratol 11: 527–531

    Google Scholar 

  • Bondy SC, Komulainen H (1988) Intracellular calcium as an index of neurotoxic damage. Toxicology 49: 35–41

    Google Scholar 

  • Carafoli E (1987) Intracellular calcium homeostasis. Ann Rev Biochem 56: 395–433

    Google Scholar 

  • Charest R, Blackmore PF, Berthon B, Exton JH (1983) Changes in free cytosolic Ca2+ in hepatocytes following αl-adrenergic stimulation: studies on quin-2-loaded hepatocytes. J Biol Chem 258: 8769–8773

    Google Scholar 

  • Cheung WY (1980) Calmodulin plays a pivotal role in cellular regulation. Science 207: 19–27

    Google Scholar 

  • Chuang D (1989) Neurotransmitter receptors and phosphoinositide turnover. Ann Rev Pharmacol Toxicol 129: 71–110

    Google Scholar 

  • DeMartino GN (1981) Calcium-dependent proteolytic activity in rat liver: identification of two proteases with different calcium requirements. Arch Biochem Biophys 211: 253–257

    Google Scholar 

  • DeMartino GN, Croall DE (1982) Calcium-dependent proteases in neuroblastoma cells. J Neurochem 38: 1642–1648

    Google Scholar 

  • Dodd PR, Hardy JA, Oakley AE, Edwardson JA, Perry EK, Delaunoy JP (1981) A rapid method for preparing synaptosomes: comparison with alternative procedures. Brain Res 226: 107–118

    Google Scholar 

  • Exton JH (1988) Mechanisms of action of calcium-mobilizing agonists: some variations on a young theme. FASEB J 2: 2670–2676

    Google Scholar 

  • Fraser AG (1986) Neurological and pulmonary adverse effects of amiodarone. Br J Clin Pract 40: 74–80

    Google Scholar 

  • Gray EG, Whittaker VP (1962) The isolation of nerve endings from brain: an electron-microscopic study of cell fragments derived by homogenization and centrifugation. J Anat (London) 96: 79–88

    Google Scholar 

  • Grynkiewicz G, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescent properties. J Biol Chem 260: 3440–3450

    Google Scholar 

  • Harris L, McKenna WJ, Rowland E, Krikler DM (1983) Side effects and possible contraindications of amiodarone use. Am Heart J 106: 916–923

    Google Scholar 

  • Hesketh TR, Smith GA, Moore JP, Taylor MV, Metcalfe JC (1983) Free cytoplasmic calcium concentration and the mitogenic stimulation of lymphocytes. J Biol Chem 258: 4876–4882

    Google Scholar 

  • Ingram DV (1983) Ocular effects in long term amiodarone therapy. Am Heart J 106: 902–905

    Google Scholar 

  • Joseph SK, Coll KE, Cooper RH, Marks JS, Williamson JR (1983) Mechanisms underlying calcium homeostasis in isolated hepatocytes. J Biol Chem 258: 731–741

    Google Scholar 

  • Kachel DL, Mayer TP, Martin WJ II (1990) Amiodarone-induced injury of human pulmonary artery endothelial cells: protection by α-tocopherol. J Pharmacol Exp Ther 254: 1107–1112

    Google Scholar 

  • Kass GEN, Wright JM, Nicotera P, Orrenius S (1988) The mechanism of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine toxicity: role of intracellular calcium. Arch Biochem Biophys 260: 789–797

    Google Scholar 

  • Kodavanti UP, Mehendale HM (1991) Amiodarone- and desethylamiodarone-induced pulmonary phospholipidosis, inhibition of phospholipases in vivo and alteration of 14C-amiodarone uptake by perfused lung. Am J Respir Cell Molec Biol 4: 369–378

    Google Scholar 

  • Komulainen H, Bondy SC (1987a) Increased free intrasynaptosomal Ca2+ by neurotoxic organometals: distinctive mechanisms. Toxicol Appl Pharmacol 88: 77–86

    Google Scholar 

  • Komulainen H, Bondy SC (1987b) Modulation of levels of free calcium within synaptosomes by organochlorine insecticides. J Pharmacol Exper Therap 241: 575–581

    Google Scholar 

  • Komulainen H, Bondy SC (1987c) The estimation of free calcium within synaptosomes and mitochondria with fura-2: comparison to quin-2. Neurochem Int 10: 55–64

    Google Scholar 

  • Li PP, White TD (1977) Rapid effects of veratradine, tetradotoxin, gramacidin D, valinomycin and NaCN on the Na+, K+ and ATP contents of synaptosomes. J Neurochem 28: 967–975

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin-phenol reagent. J Biol Chem 193: 265–275

    Google Scholar 

  • MacDermott AB, Mayer ML, Westbrook GL, Smith SJ, Barker JL (1986) NMDA-receptor activation increases cytoplasmic calcium concentration in cultured spinal cord neurons. Nature 321: 519–522

    Google Scholar 

  • Martin WJ II, Rosenow EC III (1988) Amiodarone pulmonary toxicity. Recognition and pathogenesis (Part 1). Chest 93: 1067–1075

    Google Scholar 

  • Monahan JB, Michel J (1987) Identification and characterization of an N-methyl-D-aspartate specific L-[3H]-glutamate recognition site in synaptic plasma membranes. J Neurochem 48: 1699–1708

    Google Scholar 

  • Moore L, Chen T, Knapp HR Jr, London EL (1975) Energy-dependent calcium sequestration activity in rat liver microsomes. J Biol Chem 250: 4562–4568

    Google Scholar 

  • Nicotera P, McConkey D, Svensson SA, Bellomo G, Orrenius S (1988) Correlation between cytosolic Ca 2+ concentration and cytotoxicity in hepatocytes exposed to oxidative stress. Toxicology 52: 55–63

    Google Scholar 

  • Olsen R, Santone K, Melder D, Oakes SG, Abraham R, Powis G (1988) An increase in intracellular free Ca2+ associated with serum-free growth stimulation of Swiss 3T3 fibroblasts by epidermal growth factor in the presence of bradykinin. J Biol Chem 263: 18 030–18 035

    Google Scholar 

  • Palakurthy PR, Iyer V, Meckler RJ (1987) Unusual neurotoxicity associated with amiodarone therapy. Arch Intern Med 147: 881–884

    Google Scholar 

  • Pellissier JF, Pouget J, Cross D, De Victor B, Serratrice G, Toga M (1984) Peripheral neuropathy induced by amiodarone chlorhydrate: a clinicopathological study. J Neurol Sci 63: 251–266

    Google Scholar 

  • Powis G, Olsen R, Standing JE, Kachel D, Martin WJ II (1990) Amiodarone-mediated increase in intracellular free Ca2+ associated with cellular injury to human pulmonary artery endothelial cells. Toxicol Appl Pharmacol 103: 156–164

    Google Scholar 

  • Prasada Rao KS, Fernando JCR, Ho IK, Mehendale HM (1986a) Neurotoxicity in rats following subchronic amiodarone treatment. Res Commun Chem Pathol Pharmacol 52: 217–224

    Google Scholar 

  • Prasada Rao KS, Rao SB, Camus Ph, Mehendale HM (1986b) Effect of amiodarone on Na+, K+-ATPase and Mg2+-ATPase activities in rat brain synaptosomes. Cell Biochem Funct 4: 143–151

    Google Scholar 

  • Putney JW (1987) Calcium-mobilizing receptors. Trends Pharmacol Sci 8: 481–486

    Google Scholar 

  • Rakita L, Sobol S, Mostow N, Vrobel T (1983) Amiodarone pulmonary toxicity. Am Heart J 106: 906–916

    Google Scholar 

  • Rasmussen H (1986a) The calcium messenger system. Part I. N Engl J Med 314: 1094–1101

    Google Scholar 

  • Rasmussen H (1986b) The calcium messenger system. Part II. N Engl J Med 314: 1164–1176

    Google Scholar 

  • Rasmussen H, Barrett PG (1984) Calcium messenger system: An integrated view. Physiol Rev 64: 938–984

    Google Scholar 

  • Siesjo BK (1989) Calcium and cell death. Magnesium 8: 223–237

    Google Scholar 

  • Sistare FD, Picking RA, Haynes RC Jr (1985) Sensitivity of the response of cytosolic calcium in quin-2-loaded rat hepatocytes to glucagon, adenine nucleosides, and adenine nucleotides. J Biol Chem 260: 12744–12747

    Google Scholar 

  • Sogol PB, Hershman JM, Reed AW, Dillman WH (1983) The effects of amiodarone on serum thyroid hormones and hepatic thyroxine 5′-mono-deiodination in rats. Endocrinology 113: 1464–1469

    Google Scholar 

  • Stauderman KA, Harris GD, Lovenberg W (1988) Characterization of inositol 1,4,5-trisphosphate stimulated calcium release from rat cerebellar microsomal fractions. Biochem J 255: 677–683

    Google Scholar 

  • Tsien RY, Pozzan T, Rink TJ (1982) Calcium homeostasis in intact lymphocytes: cytoplasmic free calcium monitored with a new intracellularly trapped fluorescent indicator. J Cell Biol 94: 325–334

    Google Scholar 

  • Vig PJS, Yallapragada PR, Kodavanti PRS, Desaiah D (1991) Modulation of calmodulin properties by amiodarone and its major metabolite desethylamiodarone. Pharmacol Toxicol 68: 26–33

    Google Scholar 

  • Wong EHF, Kemp JA, Priestly A, Knight AR, Woodruff GN, Iversen LL (1986) The anticonvulsant MK-801 is a potent N-methyl-D-aspartate antagonist. Proc Natl Acad Sci USA 83: 7104–7108

    Google Scholar 

  • Young RA, Mehendale HM (1986) In vitro metabolism of amiodarone by rabbit and rat liver and small intestine. Drug Metab Dispos 14: 423–429

    Google Scholar 

  • Young RA, Mehendale HM (1987) Effect of cytochrome P-450 and flavin-containing monooxygenase modifying factors on the in vitro metabolism of amiodarone by rat and rabbit. Drug Metab Dispos 15: 511–517

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Neurology, University of Mississippi Medical Center, 2500 North State Street, 39216, Jackson, MS, USA

    Prasada Rao S. Kodavanti, Srinivas N. Pentyala, Prabhakara R. Yallapragada & Durisala Desaiah

Authors
  1. Prasada Rao S. Kodavanti
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Srinivas N. Pentyala
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Prabhakara R. Yallapragada
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Durisala Desaiah
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Send offprint requests to D. Desaiah at the above address

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kodavanti, P.R.S., Pentyala, S.N., Yallapragada, P.R. et al. Amiodarone and desethylamiodarone increase intrasynaptosomal free calcium through receptor mediated channel. Naunyn-Schmiedeberg's Arch Pharmacol 345, 213–221 (1992). https://doi.org/10.1007/BF00165739

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00165739

Key words

Search

Navigation