Intracellular association of glycine receptor with gephyrin increases its plasma membrane accumulation rate

被引:65
作者
Hanus, C [1 ]
Vannier, C [1 ]
Triller, A [1 ]
机构
[1] Ecole Normale Super, Lab Biol Cellulaire Synapse Normale & Pathol, INSERM, U497, F-75005 Paris, France
关键词
gephyrin; glycine receptor; surface insertion; clustering; videomicroscopy; neurons;
D O I
10.1523/JNEUROSCI.4380-03.2004
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
Gephyrin, a tubulin-binding protein, is the core of inhibitory postsynaptic scaffolds stabilizing glycine receptors (GlyRs) and/or GABA(A) receptors. Previous ultrastructural studies in vivo and in vitro have reported a localization of gephyrin to intracellular cisternas during development or after glycinergic denervation (Seitanidou et al., 1992; Colin et al., 1996, 1998). These data were compatible with a traffic of this cytoplasmic, but membrane-associated, protein together with membrane proteins such as GlyR after exocytosis and/or endocytosis pathways. We have now investigated the consequences of a GlyR-gephyrin interaction on the localization and the dynamics of these two molecules in African green monkey kidney cells (COS-7) cells and in neurons transfected with green fluorescent protein-tagged-gephyrin and myc-tagged GlyR alpha(1) subunits. In these experiments, myc-tagged GlyR alpha(1) contained, or did not contain, the gephyrin-binding sequence (betagb) of the GlyR beta subunit. We report here that GlyR-gephyrin interaction localizes gephyrin to GlyR-containing organelles. Videomicroscopy and nocodazole treatment indicate that the movements of these vesicles are microtubule dependent. Expressing GlyR alpha(1) with a thrombin cleavage site between the myc-tag and the N-terminal of the GlyR alpha(1) subunit (Rosenberg et al., 2001) allowed monitoring of newly inserted receptors in the cell surface. Using temperature changes to block GlyR in, and then release it from, the trans-Golgi network, we show that gephyrin accelerates the accumulation of GlyR at the cell surface. Therefore, our data strongly suggest that some GlyR clusters are associated with gephyrin on their way to the cell surface and that this association increases the accumulation of GlyR at the plasma membrane.
引用
收藏
页码:1119 / 1128
页数:10
相关论文
共 58 条
[51]   LATERAL DIFFUSION IN AN ARCHIPELAGO - SINGLE-PARTICLE DIFFUSION [J].
SAXTON, MJ .
BIOPHYSICAL JOURNAL, 1993, 64 (06) :1766-1780
[52]   Single-particle tracking: The distribution of diffusion coefficients [J].
Saxton, MJ .
BIOPHYSICAL JOURNAL, 1997, 72 (04) :1744-1753
[53]   Postsynaptic organization and regulation of excitatory synapses [J].
Scannevin, RH ;
Huganir, RL .
NATURE REVIEWS NEUROSCIENCE, 2000, 1 (02) :133-141
[54]  
SEITANIDOU T, 1992, J NEUROSCI, V12, P116
[55]   Unitary assembly of presynaptic active zones from Piccolo-Bassoon transport vesicles [J].
Shapira, M ;
Zhai, RG ;
Dresbach, T ;
Bresler, T ;
Torres, VI ;
Gundelfinger, ED ;
Ziv, NE ;
Garner, CC .
NEURON, 2003, 38 (02) :237-252
[56]  
Toomre D, 1999, J CELL SCI, V112, P21
[57]   GABAA-receptor-associated protein links GABAA receptors and the cytoskeleton [J].
Wang, HB ;
Bedford, FK ;
Brandon, NJ ;
Moss, SJ ;
Olsen, RW .
NATURE, 1999, 397 (6714) :69-72
[58]   Assembling the presynaptic active zone: A characterization of an active zone precursor vesicle [J].
Zhai, RG ;
Vardinon-Friedman, H ;
Cases-Langhoff, C ;
Becker, B ;
Gundelfinger, ED ;
Ziv, NE ;
Garner, CC .
NEURON, 2001, 29 (01) :131-143