An AnkyrinG-Binding Motif Is Necessary and Sufficient for Targeting Nav1.6 Sodium Channels to Axon Initial Segments and Nodes of Ranvier

被引:111
作者
Gasser, Andreas [1 ,2 ,3 ]
Ho, Tammy Szu-Yu [4 ]
Cheng, Xiaoyang [1 ,2 ,3 ]
Chang, Kae-Jiun [4 ]
Waxman, Stephen G. [1 ,2 ,3 ]
Rasband, Matthew N. [4 ,5 ]
Dib-Hajj, Sulayman D. [1 ,2 ,3 ]
机构
[1] Yale Univ, Sch Med, Dept Neurol, New Haven, CT 06510 USA
[2] Yale Univ, Sch Med, Ctr Neurosci & Regenerat Res, New Haven, CT 06510 USA
[3] Vet Adm Connecticut Healthcare Syst, Rehabil Res Ctr, West Haven, CT 06516 USA
[4] Baylor Coll Med, Program Dev Biol, Houston, TX 77030 USA
[5] Baylor Coll Med, Dept Neurosci, Houston, TX 77030 USA
关键词
BETA-1; SUBUNITS; PROTEIN-KINASE; NEURONS; LOCALIZATION; ORGANIZATION; MYELINATION; NEUROFASCIN; CONTRIBUTES; ADHESION; MUTATION;
D O I
10.1523/JNEUROSCI.5434-11.2012
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
Neurons are highly polarized cells with functionally distinct axonal and somatodendritic compartments. Voltage-gated sodium channels Na(v)1.2 and Na(v)1.6 are highly enriched at axon initial segments (AISs) and nodes of Ranvier, where they are necessary for generation and propagation of action potentials. Previous studies using reporter proteins in unmyelinated cultured neurons suggest that an ankyrinG-binding motif within intracellular loop 2 (L2) of sodium channels is sufficient for targeting these channels to the AIS, but mechanisms of channel targeting to nodes remain poorly understood. Using a CD4-Na(v)1.2/L2 reporter protein in rat dorsal root ganglion neuron-Schwann cell myelinating cocultures, we show that the ankyrinG-binding motif is sufficient for protein targeting to nodes of Ranvier. However, reporter proteins cannot capture the complexity of full-length channels. To determine how native, full-length sodium channels are clustered in axons, and to show the feasibility of studying these channels in vivo, we constructed fluorescently tagged and functional mouse Na(v)1.6 channels for in vivo analysis using in utero brain electroporation. We show here that wild-type tagged-Na(v)1.6 channels are efficiently clustered at nodes and AISs in vivo. Furthermore, we show that mutation of a single invariant glutamic acid residue (E1100) within the ankyrinG-binding motif blocked Na(v)1.6 targeting in neurons both in vitro and in vivo. Additionally, we show that caseine kinase phosphorylation sites within this motif, while not essential for targeting, can modulate clustering at the AIS. Thus, the ankyrinG-binding motif is both necessary and sufficient for the clustering of sodium channels at nodes of Ranvier and the AIS.
引用
收藏
页码:7232 / 7243
页数:12
相关论文
共 36 条
[1]   Physiological roles of axonal ankyrins in survival of premyelinated axons and localization of voltage-gated sodium channels [J].
Bennett, V ;
Lambert, S .
JOURNAL OF NEUROCYTOLOGY, 1999, 28 (4-5) :303-318
[2]   Sodium channel Nav 1.6 is expressed along nonmyelinated axons and it contributes to conduction [J].
Black, JA ;
Renganathan, M ;
Waxman, SG .
MOLECULAR BRAIN RESEARCH, 2002, 105 (1-2) :19-28
[3]   Compact myelin dictates the differential targeting of two sodium channel isoforms in the same axon [J].
Boiko, T ;
Rasband, MN ;
Levinson, SR ;
Caldwell, JH ;
Mandel, G ;
Trimmer, JS ;
Matthews, G .
NEURON, 2001, 30 (01) :91-104
[4]   Protein kinase CK2 contributes to the organization of sodium channels in axonal membranes by regulating their interactions with ankyrin G [J].
Brechet, Aline ;
Fache, Marie-Pierre ;
Brachet, Anna ;
Ferracci, Geraldine ;
Baude, Agnes ;
Irondelle, Marie ;
Pereira, Sandrine ;
Leterrier, Christophe ;
Dargent, Benedicte .
JOURNAL OF CELL BIOLOGY, 2008, 183 (06) :1101-1114
[5]   MUTATION OF A NEW SODIUM-CHANNEL GENE, SCN8A, IN THE MOUSE MUTANT MOTOR END-PLATE DISEASE [J].
BURGESS, DL ;
KOHRMAN, DC ;
GALT, J ;
PLUMMER, NW ;
JONES, JM ;
SPEAR, B ;
MEISLER, MH .
NATURE GENETICS, 1995, 10 (04) :461-465
[6]   Nodes of Ranvier and axon initial segments are ankyrin G-dependent domains that assemble by distinct mechanisms [J].
Dzhashiashvili, Yulia ;
Zhang, Yanqing ;
Galinska, Jolanta ;
Lam, Isabel ;
Grumet, Martin ;
Salzer, James L. .
JOURNAL OF CELL BIOLOGY, 2007, 177 (05) :857-870
[7]   A sodium channel mutation linked to epilepsy increases ramp and persistent current of Nav1.3 and induces hyperexcitability in hippocampal neurons [J].
Estacion, Mark ;
Gasser, Andreas ;
Dib-Hajj, Sulayman D. ;
Waxman, Stephen G. .
EXPERIMENTAL NEUROLOGY, 2010, 224 (02) :362-368
[8]   Endocytotic elimination and domain-selective tethering constitute a potential mechanism of protein segregation at the axonal initial segment [J].
Fache, MP ;
Moussif, A ;
Fernandes, F ;
Giraud, P ;
Garrido, JJ ;
Dargent, B .
JOURNAL OF CELL BIOLOGY, 2004, 166 (04) :571-578
[9]   Axonal Sodium-Channel Bands Shape the Response to Electric Stimulation in Retinal Ganglion Cells [J].
Fried, Shelley I. ;
Lasker, Aaron C. W. ;
Desai, Neal J. ;
Eddington, Donald K. ;
Rizzo, Joseph F., III .
JOURNAL OF NEUROPHYSIOLOGY, 2009, 101 (04) :1972-1987
[10]   A targeting motif involved in sodium channel clustering at the axonal initial segment [J].
Garrido, JJ ;
Giraud, P ;
Carlier, E ;
Fernandes, F ;
Moussif, A ;
Fache, MP ;
Debanne, D ;
Dargent, B .
SCIENCE, 2003, 300 (5628) :2091-2094