Glial glutamate transporters limit spillover activation of presynaptic NMDA receptors and influence synaptic inhibition of Purkinje neurons.
Glutamate transporters limit cross talk between excitatory synapses by removing synaptically released glutamate. However, the role of glutamate transporters in limiting the action of synaptically released glutamate at inhibitory synapses remains unknown. Single and paired whole-cell patch-clamp recordings were obtained from Purkinje neurons and Bergmann glia in mouse cerebellar slices to determine the function of neuronal and glial glutamate transporters at GABAergic synapses onto Purkinje neurons. NMDA and inhibition of glutamate transporters with DL-threo-beta-benzyloxyaspartic acid (TBOA) increased the frequency of miniature IPSCs (mIPSCs) by enhancing presynaptic NMDA receptor (NMDAR) activation, whereas inhibition of cystine-glutamate antiporters had no effect on mIPSCs. Selective inhibition of glutamate transporters in an individual Bergmann glial cell enhanced mIPSC frequency recorded in an adjacent Purkinje neuron significantly more than did postsynaptic transporter inhibition. TBOA did not change the frequency of spontaneous IPSCs (sIPSCs) but decreased their amplitude, as well as that of evoked IPSCs (eIPSCs), and enhanced the paired-pulse ratio. The action of NMDAR activation on eIPSCs but not that on mIPSCs was prevented by 100 microm ryanodine. Repetitive stimulations of climbing fibers resulted in a NMDAR-dependent reduction of sIPSC amplitude, and this effect was enhanced by TBOA even when postsynaptic glutamate transporters were blocked. These data indicate that glial glutamate transporters limit glutamate spillover from excitatory to inhibitory synapses and strongly influence synaptic inhibition of Purkinje neurons by controlling NMDAR activation on GABAergic terminals.
Huang
Hao
H
Department of Neurosurgery, Yale University, New Haven, Connecticut 06520-8082, USA.
Bordey
Angélique
A
D000200:Action Potentials; D027322:Amino Acid Transport System X-AG / Q000037:antagonists & inhibitors / Q000502:physiology*; D000818:Animals; D017920:Antiporters / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002531:Cerebellum / Q000378:metabolism; D003553:Cystine / Q000378:metabolism; D005071:Evoked Potentials / Q000187:drug effects; D018698:Glutamic Acid / Q000378:metabolism; D066298:In Vitro Techniques; D051379:Mice; D009412:Nerve Fibers / Q000378:metabolism; D009433:Neural Inhibition / Q000502:physiology*; D009457:Neuroglia / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D011689:Purkinje Cells / Q000378:metabolism / Q000502:physiology*; D016194:Receptors, N-Methyl-D-Aspartate / Q000819:agonists*; D017661:Receptors, Presynaptic / Q000819:agonists*; D012433:Ryanodine / Q000494:pharmacology; D009435:Synaptic Transmission / Q000502:physiology*; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
D027322:Amino Acid Transport System X-AG; D017920:Antiporters; D016194:Receptors, N-Methyl-D-Aspartate; D017661:Receptors, Presynaptic; D012433:Ryanodine; D018698:Glutamic Acid; D003553:Cystine; D005680:gamma-Aminobutyric Acid; D002118:Calcium