title: Caffeine mediates cation influx and intracellular Ca2+ release in leech P neurones.
abstract: We investigated the effect of caffeine on the intracellular free Ca2+ concentration ([Ca2+]i) of leech P neurones by using the fluorescent indicator Fura-2. Caffeine induced a [Ca2+]i increase that was strongly reduced, but not abolished, in Ca(2+)-free solution. The effect of caffeine on [Ca2+]i was dose-dependent: while 5 mM caffeine evoked a persistent [Ca2+]i increase that could be elicited repetitively, 10 mM caffeine or more induced a transient [Ca2+]i increase that was strongly reduced upon subsequent applications at the same concentration. Surprisingly, the cells remained fully responsive to a moderately increased caffeine concentration. The caffeine-induced [Ca2+]i increase was not blocked by millimolar concentrations of La3+, Mg2+, Cd2+, Zn2+, Co2+, Ni2+, or Mn2+. While La3+ and Mg2+ had no effect on the caffeine response, the other cations caused irreversible changes in the Fura-2 fluorescence. The inhibitors of intracellular Ca2+ pumps-thapsigargin, cyclopiazonic acid (CPA), and 2,5-di-(t-butyl)-1,4-hydroquinone (BHQ)--had no effect on the caffeine-induced [Ca2+]i increase at normal extracellular Ca2+ concentration, but they reduced it in Ca(2+)-free solution. Ryanodine had no effect on the caffeine-induced [Ca2+]i increase at normal extracellular Ca2+ concentration, and also in Ca(2+)-free solution it seemed to be largely ineffective. Caffeine evoked complete fluctuations of the membrane potential. The effect in Ca2+ free and in Na(+)-free solution suggests that the depolarizing response components were mainly due to Na+ influx, while Ca2+ reduced the Na+ influx and/or activated mechanisms which re- or hyperpolarize the cells. It is concluded that leech P neurones possess caffeine-sensitive intracellular Ca2+ stores, as well as caffeine-sensitive ion channels, in the plasma membrane that are activated by a voltage-independent mechanism. The plasma membrane channels are permeable to various divalent cations including Ca2+, and possibly also to Na+.
authors.affiliation: Institut für Neurobiologie, Heinrich-Heine-Universität Düsseldorf, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Schoppe
authors.forename: P
authors.initials: P
authors.lastname: Hochstrate
authors.forename: W R
authors.initials: WR
authors.lastname: Schlue
mesh_terms: D000818:Animals; D002110:Caffeine / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism*; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors; D002412:Cations; D015226:Dinucleoside Phosphates / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006873:Hydroquinones / Q000494:pharmacology; D007424:Intracellular Fluid; D007865:Leeches; D008564:Membrane Potentials / Q000187:drug effects; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D012433:Ryanodine / Q000494:pharmacology; D012964:Sodium / Q000494:pharmacology; D012996:Solutions; D019284:Thapsigargin / Q000494:pharmacology
chemical_list: D002412:Cations; D015226:Dinucleoside Phosphates; D004791:Enzyme Inhibitors; D006873:Hydroquinones; D012996:Solutions; D012433:Ryanodine; C013907:cytidylyl adenosine; D002110:Caffeine; D019284:Thapsigargin; D012964:Sodium; C018855:2-tert-butylhydroquinone; D000252:Calcium-Transporting ATPases; D002118:Calcium