EC Neurology

Research Article Volume 12 Issue 7 - 2020

Mobilization of Intracellular Calcium by Ryanodine in Cultured, Identified Molluscan Neurons is Cell Specific

IA Ahmed1, PM Hopkins2,3 and W Winlow4,5 *

1 Faculty of Medicine, University of Garden City, Khartoum, Sudan
2 Leeds Institute of Medical Research at St James’s, University of Leeds, Leeds, United Kingdom
3 Malignant Hyperthermia Investigation Unit, St. James’s University Hospital, Leeds, United Kingdom
4 Department of Biology, University of Naples Federico II, Via Cintia, Naples, Italy
5 Institute of Ageing and Chronic Diseases, University of Liverpool, Liverpool, United Kingdom

*Corresponding Author: William Winlow, Department of Biology, University of Naples Federico II, Via Cintia, Naples, Italy and Institute of Ageing and Chronic Diseases, University of Liverpool, Liverpool, United Kingdom.
Received: June 11, 2020; Published: June 30, 2020



  • Previously we showed that the calcium mobilizing agent, ryanodine, altered action potential shape in some identified neurons of the mollusc Lymnaea stagnalis.
  • Here we investigate the actions of ryanodine (1 μM and 5 μM) on intracellular calcium concentration [Ca2+] i in the same identified neurons in single cell culture.
  • The free [Ca2+] i levels in these neurons were determined using the cell permeable Ca2+ indicator Fura-2AM, both in the pres- ence and absence of extracellular calcium (0-Ca2+/EGTA).
  • In the presence of extracellular calcium, both concentrations of ryanodine produced a substantial increase in [Ca2+] i in three neuron types (VV1or 2, RPD1 and RPD2), whereas, in the fourth neuron type (RPeD1) there was a decrease below resting level.
  • The free [Ca2+] i remained elevated even after washing with normal saline, except in one case (VV1or 2) where it was found to be reversible. However, there was a general decline in the level of [Ca2+] i after addition of ryanodine in the absence of Ca2+ in the external medium.
  • Therefore, extracellular Ca2+ is a prerequisite for any appreciable rise in [Ca2+] i due to ryanodine.

keywords:Ryanodine; Intracellular Calcium Concentration; Lymnaea Neurons; Neuron Specificity

  1. Yamaguchi N. “Molecular insights into calcium dependent regulation of ryanodine receptor calcium release channels. (In: Islam M. (editions) Calcium Signaling, 2nd Edition)”. Advances in Experimental Medicine and Biology 1131 (2019): 321-336.
  2. Miller RM. “The control of neuronal Ca2+ homeostasis”. Progress in Neurobiology 37 (1991): 255-285.
  3. Sandler VM and Barbara JG. “Calcium-induced calcium release contributes to action potential-evoked calcium transients in hippocampal CA1 pyramidal neurons”. The Journal of Neuroscience 19 (1999): 4325-4336.
  4. Petrou T., et al. “Intracellular calcium mobilization in response to ion channel regulators via a calcium-induced calcium release mechanism”. Journal of Pharmacology and Experimental Therapeutics 360 (2017): 378-387.
  5. Lanner JT., et al. “Ryanodine receptors: structure, expression, molecular details, and function in calcium release”. Cold Spring Harbor Perspectives in Biology 2 (2010): a003996.
  6. Dunn TW and Syed NI. “Ryanodine receptor-transmitter release site coupling increases quantal size in a synapse-specific manner”. European Journal of Neuroscience 24 (2006): 1591-1605.
  7. Dunn TW., et al. “An action potential- induced and ryanodine sensitive calcium transient dynamically regulates transmitter release between Lymnaea neurons”. Synapse 63 (2009): 61-68.
  8. Ahmed IA., et al. “Low concentrations of caffeine raise intracellular calcium concentration only in the presence of extracellular calcium in cultured molluscan neurons”. General Pharmacology 28 (1997): 245-250.
  9. Winlow W and Polese G. “A Neuroplastic Network Underlying Behaviour and Seasonal Change in Lymnaea stagnalis: A Neuroecological Standpoint”. In Neuroecology and Neuroethology in Molluscs: The Interface Between Behaviour and Environment (2014): 145-176.
  10. Winlow W., et al. “Sense and Insensibility - Appraisal of the effects of clinical anesthetics on gastropod and cephalopod molluscs as a step to improved welfare in Cephalopods”. Frontiers in Physiology 1147 (2018): 24.
  11. Moghadam HF and Winlow W. “Pharmacological Dissection of Potassium Currents in Cultured, Identified, Molluscan Neurons”. EC Neurology 11 (2019): 346-356.
  12. Benjamin PR and Winlow W. “The distribution of three wide-acting synaptic inputs to identified neurons in the isolated brain of Lymnaea stagnalis (L)”. Comparative Biochemistry and Physiology B 70A (1981): 293-307.
  13. Qazzaz MM and Winlow W. “Differential Actions of Volatile Anaesthetics and a Systemic Barbiturate on Strongly Electrically Coupled Neurons”. EC Neurology4 (2015):188-204.
  14. Qazzaz MM and Winlow W. “Modulation of the passive membrane properties of a pair of strongly electrically coupled neurons by anaesthetics”. EC Neurology4 (2017): 187-200.
  15. Walcourt-Ambakederemo A and Winlow W. “5-HT receptors on identified Lymnaea neurones in culture. Pharmacological characterization of 5-HT1A receptors”. Comparative Biochemistry and Physiology B 107C (1994): 129-141.
  16. Grynkiewicz G., et al. “A new generation of Ca2+ indicators with greatly improved fluorescent properties”. Journal of Biological Chemistry 260 (1985): 3440-3450.
  17. Ahern GP., et al. “Single channel activity of the ryanodine receptor calcium release channel is modulated by FK-506”. FEBS Letters 352 (1994): 369-374.
  18. Ahmed IA., et al. “Caffeine and ryanodine modify the half-width of somatic action potentials in identified molluscan neurons”. The Journal of Physiology 459 (1993): 13P.
  19. Ahmed IA., et al. “Caffeine and ryanodine differentially modify a calcium-dependent component of soma action potentials in identified molluscan neurones in situ”. Comparative Biochemistry and Physiology B 105C (1993): 363-372.
  20. Haydon PG., et al. “The effects of menthol on central neurones of the pond-snail, Lymnaea stagnalis (L)”. Comparative Biochemistry and Physiology B 73C (1982): 95-100.
  21. Girdlestone D., et al. “The actions of halothane on spontaneous activity, action potential shape and synaptic connections of the giant serotonin-containing neurones of Lymnaea stagnalis (L). Comp”. Biochemistry and Physiology 93C (1989): 333-339.
  22. Blatz AL and Magleby KL. “Calcium-activated potassium channels”. Trends in Neuroscience 10 (1987): 463-467.
  23. Marty A. “The physiological role of calcium-dependent channels”. Trends in Neuroscience 12 (1989): 420-424.
  24. Meech RW. “Membranes, gates and channels”. In: The Mollusca, 9 neurobiology and Behavior, Part 2 (edition: Willows, A.O.D.), (1986):189-277.
  25. Zhang Y., et al. “Prolonged activation of Ca2+-activated K+ current contributes to the long-lasting refractory period of Aplysia bag cell neurons”. Trends in Neuroscience 22 (2002): 10134-10141.
  26. Moghadam H Fathi., et al. “A comparative study of the specific effects of systemic and volatile anesthetics on identified neurons of Lymnaea stagnalis (L.), both in the isolated brain and in single cell culture”. Frontiers in Physiology 10 (2019): 583.
  27. Faber ESL and Sah P. “Ca2+-activated K+ (BK) channel inactivation contributes to spike broadening during repetitive firing in the rat lateral amygdala”. The Journal of Physiology 2 (2003): 483-497.
  28. Sah P and Faber ESL. “Channels underlying neuronal calcium-activated potassium current”. Progress in Neurobiology 66 (2002): 345- 353.
  29. Bean BP. “The action potential in mammalian central neurons”. Nature Reviews - Neuroscience 8 (2007): 451-465.
  30. Wang B., et al. “Knockout of the BK β4-subunit promotes a functional coupling of BK channels and ryanodine receptors that mediate a f-AHP-induced increase in excitability”. Journal of Neurophysiology 116 (2016): 456-465.
  31. Irie T and Trussell LO. “Double nanodomain coupling of calcium channels, ryanodine receptors and BK channels controls generation of burst firing”. Neuron4 (2017): 856-870.
  32. Kim D., et al. “Coupled positive and negative feedback circuits form an essential building block of cellular signaling pathways”. Bio Essays 29 (2006): 85-90.
  33. Beard NA., et al. “Calsequestrin is an inhibitor of skeletal muscle ryanodine receptor calcium release channels”. Biophysical Journal 82 (2002): 310-320.
  34. Tripathy A., et al. “Calmodulin activation and inhibition of skeletal muscle Ca2+ release channel (ryanodine receptor)”. Biophysical Journal 69 (1995): 106-119.

W Winlow., et al. “Mobilization of Intracellular Calcium by Ryanodine in Cultured, Identified Molluscan Neurons is Cell Specific”. EC Neurology  12.7 (2020): 67-75.

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