In Parkinson's disease, the protein α-synuclein (αS) is produced within neurons and also appears in the extracellular fluid. In this study in hippocampal neurons, αS formed non-selective cation channels with multiple levels of conductance and rectification depending on their insertion site. αS channels induced local spontaneous increases in intracellular Na(+) and Ca(2+), depolarized neurons, augmented bursting activity and stimulated the opening of ATP-sensitive K(+) channels. Non-selective channels were also observed in neurons transfected with either wild-type or mutant A53T αS, and after extracellular application of these proteins. The properties of αS channels in neuronal membranes suggest that extracellular αS is more toxic than αS produced within neurons. In Parkinson's disease and several other neurodegenerative diseases, the protein α-synuclein (αS) is produced within neurons and accumulates in the extracellular fluid. Several mechanisms of αS action are proposed, one of which is the formation of cation-permeable pores that may mediate toxicity. αS induces non-selective cation channels in lipid bilayers, but whether this occurs in living neurons and which properties the channels possess have not yet been examined. In this study the properties of αS channels in dissociated hippocampal neurons are documented. In cell-attached recordings the incorporation of αS into membranes was driven by applied negative potentials. These channels exhibited multiple levels of conductance (30, 70 and 120 pS at -100 mV) and inward rectification. The persistent activity of αS channels induced local changes in intracellular Na(+) and Ca(2+), depolarized neurons and augmented bursting activity. αS channels formed by adding αS to the intracellular membrane in inside-out patches exhibited outward rectification. αS channels were equally permeable to Na(+), K(+) and Ca(2+). These channels were also observed in neurons transfected with wild-type or mutant A53T αS, and after extracellular application of wild-type or mutant A53T αS proteins. Opening of αS channels stimulated opening of ATP-sensitive K(+) (KATP ) channels and did not interfere with the activity of delayed rectifier K(+) channels. The properties of αS channels in neuronal membranes suggest stronger toxicity of extracellularly applied αS than intracellular αS. Enhancement of neuronal excitability and distortions in ion homeostasis may underlie the toxic effects of αS that can be dampened by KATP channels.
© 2014 The Author. The Journal of Physiology © 2014 The Physiological Society.