title: Zn(2+) -induced Ca(2+) release via ryanodine receptors triggers calcineurin-dependent redistribution of cortical neuronal Kv2.1 K(+) channels.
abstract: Increases in intracellular Zn(2+) concentrations are an early, necessary signal for the modulation of Kv2.1 K(+) channel localization and physiological function. Intracellular Zn(2+) -mediated Kv2.1 channel modulation is dependent on calcineurin, a Ca(2+) -activated phosphatase. We show that intracellular Zn(2+) induces a significant increase in ryanodine receptor-dependent cytosolic Ca(2+) transients, which leads to a calcineurin-dependent redistribution of Kv2.1 channels from pre-existing membrane clusters to diffuse localization. As such, the link between Zn(2+) and Ca(2+) signalling in this Kv2.1 modulatory pathway is established. We observe that a sublethal ischaemic preconditioning insult also leads to Kv2.1 redistribution in a ryanodine receptor-dependent fashion. We suggest that Zn(2+) may be an early and ubiquitous signalling molecule mediating Ca(2+) release from the cortical endoplasmic reticulum via ryanodine receptor activation.
  
  
  
  Sublethal injurious stimuli in neurons induce transient increases in free intracellular Zn(2+) that are associated with regulating adaptive responses to subsequent lethal injury, including alterations in the function and localization of the delayed-rectifier potassium channel, Kv2.1. However, the link between intracellular Zn(2+) signalling and the observed changes in Kv2.1 remain undefined. In the present study, utilizing exogenous Zn(2+) treatment, along with a selective Zn(2+) ionophore, we show that transient elevations in intracellular Zn(2+) concentrations are sufficient to induce calcineurin-dependent Kv2.1 channel dispersal in rat cortical neurons in vitro, which is accompanied by a relatively small but significant hyperpolarizing shift in the voltage-gated activation kinetics of the channel. Critically, using a molecularly encoded calcium sensor, we found that the calcineurin-dependent changes in Kv2.1 probably occur as a result of Zn(2+) -induced cytosolic Ca(2+) release via activation of neuronal ryanodine receptors. Finally, we couple this mechanism with an established model for in vitro ischaemic preconditioning and show that Kv2.1 channel modulation in this process is also ryanodine receptor-sensitive. Our results strongly suggest that intracellular Zn(2+) -initiated signalling may represent an early and possibly widespread component of Ca(2+) -dependent processes in neurons.
authors.affiliation: Department of Neurobiology.|Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh School of Medicine, PA, USA.
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Schulien
authors.affiliation: Department of Neurobiology.|Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh School of Medicine, PA, USA.
authors.forename: Jason A
authors.initials: JA
authors.lastname: Justice
authors.affiliation: Department of Neurology.|Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh School of Medicine, PA, USA.
authors.forename: Roberto
authors.initials: R
authors.lastname: Di Maio
authors.affiliation: Department of Neurobiology.
authors.forename: Zachary P
authors.initials: ZP
authors.lastname: Wills
authors.affiliation: Department of Neurobiology.
authors.forename: Niyathi H
authors.initials: NH
authors.lastname: Shah
authors.affiliation: Department of Neurobiology.|Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh School of Medicine, PA, USA.
authors.forename: Elias
authors.initials: E
authors.lastname: Aizenman
mesh_terms: D000818:Animals; D019703:Calcineurin / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000187:drug effects / Q000378:metabolism*; D002712:Chlorides / Q000494:pharmacology*; D005260:Female; D008297:Male; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D019837:Ryanodine Receptor Calcium Release Channel / Q000378:metabolism*; D051669:Shab Potassium Channels / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D017967:Zinc Compounds / Q000494:pharmacology*
chemical_list: D002712:Chlorides; C496991:Kcnb1 protein, rat; D019837:Ryanodine Receptor Calcium Release Channel; D051669:Shab Potassium Channels; D017967:Zinc Compounds; C016837:zinc chloride; D019703:Calcineurin; D002118:Calcium