Heat Shock Cognate Protein 70 Regulates Gephyrin Clustering

被引:28
|
作者
Machado, Patricia [2 ]
Rostaing, Philippe [2 ]
Guigonis, Jean-Marie [3 ]
Renner, Marianne [2 ]
Dumoulin, Andrea [2 ]
Samson, Michel [3 ]
Vannier, Christian [2 ]
Triller, Antoine [1 ,2 ]
机构
[1] Ecole Normale Super, INSERM, U1024, Lab Biol Cellulaire Synapse,Inst Biol, F-75005 Paris, France
[2] CNRS, UMR 8197, F-75005 Paris, France
[3] Univ Nice Sophia Antipolis, Inst Federat Rech 50, Fac Med Pasteur, F-06107 Nice, France
来源
JOURNAL OF NEUROSCIENCE | 2011年 / 31卷 / 01期
关键词
GLYCINE RECEPTOR; POSTSYNAPTIC DENSITY; UNCOATING ATPASE; PLASMA-MEMBRANE; INHIBITORY SYNAPSES; FLUORESCENT PROTEIN; HYPERTHERMIC STRESS; CRYSTAL-STRUCTURE; ESCHERICHIA-COLI; DENDRITIC SPINES;
D O I
10.1523/JNEUROSCI.2533-10.2011
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
Formation and stabilization of postsynaptic glycine receptor (GlyR) clusters result from their association with the polymerized scaffold protein gephyrin. At the cell surface, lateral diffusion and local trapping of GlyR by synaptic gephyrin clusters is one of the main factors controlling their number. However, the mechanisms regulating gephyrin/GlyR cluster sizes are not fully understood. To identify molecular binding partners able to control gephyrin cluster stability, we performed pull-down assays with full-length or truncated gephyrin forms incubated in a rat spinal cord extract, combined with mass spectrometric analysis. We found that heat shock cognate protein 70 (Hsc70), a constitutive member of the heat shock protein 70 (Hsp70) family, selectively binds to the gephyrin G-domain. Immunoelectron microscopy of mouse spinal cord sections showed that Hsc70 could be colocalized with gephyrin at inhibitory synapses. Furthermore, ternary Hsc70-gephyrin-GlyR coclusters were formed following transfection of COS-7 cells. Upon overexpression of Hsc70 in mouse spinal cord neurons, synaptic accumulation of gephyrin was significantly decreased, but GlyR amounts were unaffected. In the same way, Hsc70 inhibition increased gephyrin accumulation at inhibitory synapses without modifying GlyR clustering. Single particle tracking experiments revealed that the increase of gephyrin molecules reduced GlyR diffusion rates without altering GlyR residency at synapses. Our findings demonstrate that Hsc70 regulates gephyrin polymerization independently of its interaction with GlyR. Therefore, gephyrin polymerization and synaptic clustering of GlyR are uncoupled events.
引用
收藏
页码:3 / 14
页数:12
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