High-affinity GABA uptake by neuronal GAT1 transporters provokes release of [(3)H]GABA by homoexchange and through GAT1-independent Ca(2+)-mediated mechanisms

Neuropharmacology. 2015 Jan:88:164-70. doi: 10.1016/j.neuropharm.2014.08.007. Epub 2014 Aug 20.

Abstract

High-affinity uptake of GABA into nerve terminals may have functions other than recapture of the neurotransmitter. Synaptosomes purified from mouse cerebellum were prelabelled with [(3)H]GABA and then superfused with GABA and drugs selective for some presynaptic targets. Influx of GABA through GAT1 transporters stimulated efflux of [(3)H]GABA in a concentration-dependent manner (EC50 ∼ 3 μM). The efflux of the transmitter occurred in part by GAT1 reversal through the so called homoexchange. The ion fluxes (particularly Na(+) influx) accompanying GABA uptake triggered intraterminal Ca(2+) signals through both plasmalemmal Na(+)/Ca(2+) exchangers, sensitive to KB-R7943 or to ifenprodil and mitochondrial Na(+)/Ca(2+) exchangers, sensitive to CGP37157. These Ca(2+) signals likely facilitated GABA release from nerve terminals via niflumic acid- and NPPB-sensitive anion channels. The results show that GABA, at concentrations corresponding to the high-affinity uptake, can evoke GABA release which occurs in part by the expected GAT1-mediated homoexchange, while the transporter-independent component of the GABA uptake-evoked GABA release takes place by hitherto unsuspected mechanisms which include Na(+)/Ca(2+) exchangers and anion channels. The significance of the novel function of the GABA high-affinity uptake here identified deserves further multidisciplinary investigation.

Keywords: Anion channels; GABA GAT1 transporter; GABA homoexchange; GABA release; Mitochondrial Na(+)/Ca(2+) exchanger; Plasmalemmal Na(+)/Ca(2+) exchanger.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / metabolism*
  • Cations / metabolism
  • Cerebellum / drug effects
  • Cerebellum / metabolism
  • GABA Plasma Membrane Transport Proteins / metabolism*
  • Ion Channels / metabolism
  • Male
  • Mice
  • Mitochondria / drug effects
  • Mitochondria / metabolism
  • Neurons / drug effects
  • Neurons / metabolism*
  • Sodium / metabolism
  • Sodium-Calcium Exchanger / metabolism
  • Synaptosomes / drug effects
  • Synaptosomes / metabolism
  • Tritium
  • gamma-Aminobutyric Acid / metabolism*

Substances

  • Cations
  • GABA Plasma Membrane Transport Proteins
  • Ion Channels
  • Slc6a1 protein, mouse
  • Sodium-Calcium Exchanger
  • Tritium
  • gamma-Aminobutyric Acid
  • Sodium
  • Calcium