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Strychnine-Blocked Glycine Receptor Is Removed from Synapses by a Shift in Insertion/Degradation Equilibrium

https://doi.org/10.1006/mcne.2001.1074Get rights and content

Abstract

The long-term inhibition by strychnine of glycine receptor activity in neurons provokes the receptor's selective intracellular accumulation and disappearance from synapses. This could result either from a disruption of the postsynaptic anchoring of the receptor or from an arrest of its exocytic transport. In this study we combined biochemical and fluorescence microscopy analyses to determine on a short time scale the fate of the strychnine-inactivated glycine receptor. Quantification of the cellular content of receptor showed that the rapid accumulation depends on protein synthesis. Cell surface biotinylation of neurons demonstrated that strychnine did not accelerate the turnover rate of the receptor. Labeling of endosomes indicated that, in strychnine-treated cells, the accumulated receptor is not blocked in the endosomal transport pathway. Taken together, these results indicate that strychnine does not destabilize the postsynaptic receptor but triggers its disappearance from synapses by a nondegradative sequestration of newly synthesized molecules in a nonendocytic compartment.

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      The direct interaction of the GlyR β-IL subunit with gephyrin seems to be involved in the postsynaptic accumulation of the GlyR (Kirsch et al., 1993; Meyer et al., 1995). The GlyR–gephyrin interaction is important for the rate of surface accumulation, postsynaptic accumulation, postsynaptic stabilization and internalization of GlyRs from synapses to intracellular organelles (Kirsch et al., 1993; Feng et al., 1998; Rasmussen et al., 2002; Hanus et al., 2004; Levi et al., 2004). Furthermore, GlyR and Ca+ channel activity has also been shown to be important for GlyR surface stability and the postsynaptic accumulation of the glycine receptor (Kirsch and Betz, 1998; Levi et al., 1998; Kneussel and Betz, 2000).

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