Acute knockdown of GABARAP by siRNA in cultured neurons has revealed a role of GABARAP in rapid NMDA-induced functional plasticity of inhibitory synapses (Marsden et al., 2007). NMDA receptor-mediated Ca2+ influx following moderate stimulation of neurons with NMDA leads to a rapid increase in the number of postsynaptic GABAAR clusters and mIPSC amplitudes (see also further below and Figure 5C). In addition to GABARAP this mechanism involves Ca2+ calmodulin-dependent kinase II (CaMKII), the vesicular trafficking factor N-ethylmaleimide-sensitive learn more factor (NSF), and glutamate receptor interacting protein (GRIP).

The rate of GABAAR endocytosis following treatment with NMDA was unaltered, suggesting that GABARAP-dependent potentiation of inhibitory synapses involves increased exocytosis rather than reduced endocytosis of GABAARs (Marsden et al., 2007). These findings represent thus far the only loss-of-function experiments showing an essential role for endogenous GABARAP in GABAAR trafficking. The relevant protein-protein interactions and CaMKII phosphorylation targets have so far not been identified. However, experiments in

heterologous cells allow speculation that this mechanism might involve CaMKII-induced phosphorylation of the β3 subunit at S383 (Houston et al., 2007). The data LY294002 price summarized thus far suggest that GABARAP promotes the regulated, activity-dependent, and CaMKII-mediated translocation of GABAARs from intracellular compartments to

the somatodendritic plasma membrane. However, a more general role of GABARAP in exocytosis of GABAARs is difficult almost to reconcile with other findings. First, GABARAP has been proposed to contribute to rebound potentiation, a neural activity-induced postsynaptic form of long-term potentiation (LTP) of inhibitory synapses on Purkinje cell neurons (Kawaguchi and Hirano, 2007). Using electrical stimulation of cultured Purkinje cells to mimic rebound potentiation, the authors found evidence that this form of plasticity is critically dependent on a CaMKII-dependent conformational alteration of GABARAP. However, LTP of GABAergic synapses occurred without measurable changes in the cellular distribution and cell surface expression of GABAARs. Given that GABARAP is absent at synapses (Kneussel et al., 2000 and Kittler et al., 2001), the exact role of GABARAP in this form of plasticity requires further clarification. Second, the aforementioned PE conjugation of GABARAP is critically involved in autophagy, an evolutionarily conserved form of bulk transport of membranes and cytoplasm to lysosomes for protein degradation (Tanida et al., 2004). Consistent with a role of GABARAP in autophagy, there is evidence that GABAARs are subject to autophagy in worms. Body wall muscle cells of C.