Gephyrin regulates GABAergic and glutamatergic synaptic transmission in hippocampal cell cultures.
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Gephyrin regulates GABAergic and glutamatergic synaptic transmission in hippocampal cell cultures. / Varley, Zeynep Kasap; Pizzarelli, Rocco; Antonelli, Roberta; Stancheva, Stefka H; Kneussel, Matthias; Cherubini, Enrico; Zacchi, Paola.
In: J BIOL CHEM, Vol. 286, No. 23, 23, 2011, p. 20942-20951.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Gephyrin regulates GABAergic and glutamatergic synaptic transmission in hippocampal cell cultures.
AU - Varley, Zeynep Kasap
AU - Pizzarelli, Rocco
AU - Antonelli, Roberta
AU - Stancheva, Stefka H
AU - Kneussel, Matthias
AU - Cherubini, Enrico
AU - Zacchi, Paola
PY - 2011
Y1 - 2011
N2 - Gephyrin is a scaffold protein essential for stabilizing glycine and GABA(A) receptors at inhibitory synapses. Here, recombinant intrabodies against gephyrin (scFv-gephyrin) were used to assess whether this protein exerts a transynaptic action on GABA and glutamate release. Pair recordings from interconnected hippocampal cells in culture revealed a reduced probability of GABA release in scFv-gephyrin-transfected neurons compared with controls. This effect was associated with a significant decrease in VGAT, the vesicular GABA transporter, and in neuroligin 2 (NLG2), a protein that, interacting with neurexins, ensures the cross-talk between the post- and presynaptic sites. Interestingly, hampering gephyrin function also produced a significant reduction in VGLUT, the vesicular glutamate transporter, an effect accompanied by a significant decrease in frequency of miniature excitatory postsynaptic currents. Overexpressing NLG2 in gephyrin-deprived neurons rescued GABAergic but not glutamatergic innervation, suggesting that the observed changes in the latter were not due to a homeostatic compensatory mechanism. Pulldown experiments demonstrated that gephyrin interacts not only with NLG2 but also with NLG1, the isoform enriched at excitatory synapses. These results suggest a key role of gephyrin in regulating transynaptic signaling at both inhibitory and excitatory synapses.
AB - Gephyrin is a scaffold protein essential for stabilizing glycine and GABA(A) receptors at inhibitory synapses. Here, recombinant intrabodies against gephyrin (scFv-gephyrin) were used to assess whether this protein exerts a transynaptic action on GABA and glutamate release. Pair recordings from interconnected hippocampal cells in culture revealed a reduced probability of GABA release in scFv-gephyrin-transfected neurons compared with controls. This effect was associated with a significant decrease in VGAT, the vesicular GABA transporter, and in neuroligin 2 (NLG2), a protein that, interacting with neurexins, ensures the cross-talk between the post- and presynaptic sites. Interestingly, hampering gephyrin function also produced a significant reduction in VGLUT, the vesicular glutamate transporter, an effect accompanied by a significant decrease in frequency of miniature excitatory postsynaptic currents. Overexpressing NLG2 in gephyrin-deprived neurons rescued GABAergic but not glutamatergic innervation, suggesting that the observed changes in the latter were not due to a homeostatic compensatory mechanism. Pulldown experiments demonstrated that gephyrin interacts not only with NLG2 but also with NLG1, the isoform enriched at excitatory synapses. These results suggest a key role of gephyrin in regulating transynaptic signaling at both inhibitory and excitatory synapses.
KW - Animals
KW - Humans
KW - Rats
KW - Rats, Wistar
KW - HEK293 Cells
KW - Synaptic Transmission/physiology
KW - Carrier Proteins/biosynthesis/genetics
KW - Cell Adhesion Molecules, Neuronal/genetics/metabolism
KW - Hippocampus/cytology/metabolism
KW - Membrane Proteins/biosynthesis/genetics
KW - Nerve Tissue Proteins/genetics/metabolism
KW - Presynaptic Terminals/metabolism
KW - Receptors, GABA-A/genetics/metabolism
KW - Receptors, Glycine/genetics/metabolism
KW - Vesicular Inhibitory Amino Acid Transport Proteins/genetics/metabolism
KW - gamma-Aminobutyric Acid/genetics/metabolism
KW - Animals
KW - Humans
KW - Rats
KW - Rats, Wistar
KW - HEK293 Cells
KW - Synaptic Transmission/physiology
KW - Carrier Proteins/biosynthesis/genetics
KW - Cell Adhesion Molecules, Neuronal/genetics/metabolism
KW - Hippocampus/cytology/metabolism
KW - Membrane Proteins/biosynthesis/genetics
KW - Nerve Tissue Proteins/genetics/metabolism
KW - Presynaptic Terminals/metabolism
KW - Receptors, GABA-A/genetics/metabolism
KW - Receptors, Glycine/genetics/metabolism
KW - Vesicular Inhibitory Amino Acid Transport Proteins/genetics/metabolism
KW - gamma-Aminobutyric Acid/genetics/metabolism
M3 - SCORING: Journal article
VL - 286
SP - 20942
EP - 20951
JO - J BIOL CHEM
JF - J BIOL CHEM
SN - 0021-9258
IS - 23
M1 - 23
ER -
