Advertisement

Functional Interaction between the Scaffold Protein Kidins220/ARMS and Neuronal Voltage-Gated Na+ Channels*

  • Fabrizia Cesca
    Correspondence
    To whom correspondence should be addressed: Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genova, Italy. Tel.: 39-010-71781-788; Fax: 39-010-71781-230;
    Affiliations
    Center for Synaptic Neuroscience, Istituto Italiano di Tecnologia, 16163 Genova, Italy
    Search for articles by this author
  • Annyesha Satapathy
    Affiliations
    Center for Synaptic Neuroscience, Istituto Italiano di Tecnologia, 16163 Genova, Italy

    Department of Experimental Medicine, University of Genova, 16132 Genova, Italy
    Search for articles by this author
  • Enrico Ferrea
    Affiliations
    Center for Synaptic Neuroscience, Istituto Italiano di Tecnologia, 16163 Genova, Italy

    Sensorimotor Group, German Primate Center, 37077 Göttingen, Germany
    Search for articles by this author
  • Thierry Nieus
    Affiliations
    Neuro Technology Department, Istituto Italiano di Tecnologia, 16163 Genova, Italy
    Search for articles by this author
  • Fabio Benfenati
    Affiliations
    Center for Synaptic Neuroscience, Istituto Italiano di Tecnologia, 16163 Genova, Italy

    Department of Experimental Medicine, University of Genova, 16132 Genova, Italy
    Search for articles by this author
  • Joachim Scholz-Starke
    Correspondence
    To whom correspondence should be addressed: Institute of Biophysics, Consiglio Nazionale delle Ricerche, Via de Marini 6, 16149 Genova, Italy. Tel.: 39-010-6475593; Fax: 39-010-6475500;
    Affiliations
    Center for Synaptic Neuroscience, Istituto Italiano di Tecnologia, 16163 Genova, Italy

    Institute of Biophysics, Consiglio Nazionale delle Ricerche, 16149 Genova, Italy
    Search for articles by this author
  • Author Footnotes
    * This work was supported by grants from the Compagnia di San Paolo (to F. C.), Istituto Italiano di Tecnologia (to F. B.), and European Union FP7 Integrating Project “Desire” Grant 602531 (to F. B. and F. C.). The authors declare that they have no conflict of interest with the contents of this article.
Open AccessPublished:June 02, 2015DOI:https://doi.org/10.1074/jbc.M115.654699
      Kidins220 (kinase D-interacting substrate of 220 kDa)/ankyrin repeat-rich membrane spanning (ARMS) acts as a signaling platform at the plasma membrane and is implicated in a multitude of neuronal functions, including the control of neuronal activity. Here, we used the Kidins220−/− mouse model to study the effects of Kidins220 ablation on neuronal excitability. Multielectrode array recordings showed reduced evoked spiking activity in Kidins220−/− hippocampal networks, which was compatible with the increased excitability of GABAergic neurons determined by current-clamp recordings. Spike waveform analysis further indicated an increased sodium conductance in this neuronal subpopulation. Kidins220 association with brain voltage-gated sodium channels was shown by co-immunoprecipitation experiments and Na+ current recordings in transfected HEK293 cells, which revealed dramatic alterations of kinetics and voltage dependence. Finally, an in silico interneuronal model incorporating the Kidins220-induced Na+ current alterations reproduced the firing phenotype observed in Kidins220−/− neurons. These results identify Kidins220 as a novel modulator of Nav channel activity, broadening our understanding of the molecular mechanisms regulating network excitability.

      Introduction

      The scaffold protein Kidins220
      The abbreviations used are: Kidins220
      kinase D-interacting substrate of 220 kDa
      ARMS
      ankyrin repeat-rich membrane spanning
      Nav channel
      voltage-gated Na+ channel
      AP
      action potential
      Trk
      tropomyosin-related kinase
      MEA
      multielectrode array
      si-Kids
      Nav1.2/Kidins220-expressing cells showing slow inactivation kinetics
      fi-Kids
      Nav1.2/Kidins220-expressing cells showing fast inactivation kinetics.
      /ARMS (
      • Iglesias T.
      • Cabrera-Poch N.
      • Mitchell M.P.
      • Naven T.J.
      • Rozengurt E.
      • Schiavo G.
      Identification and cloning of Kidins220, a novel neuronal substrate of protein kinase D.
      ,
      • Kong H.
      • Boulter J.
      • Weber J.L.
      • Lai C.
      • Chao M.V.
      An evolutionarily conserved transmembrane protein that is a novel downstream target of neurotrophin and ephrin receptors.
      ), hereafter referred to as Kidins220, acts as a signaling platform implicated in many cellular functions through a plethora of interactions with membrane receptors, cytosolic signaling components, and cytoskeletal proteins (
      • Neubrand V.E.
      • Cesca F.
      • Benfenati F.
      • Schiavo G.
      Kidins220/ARMS as a functional mediator of multiple receptor signalling pathways.
      ). In the nervous system, where the protein is preferentially expressed, Kidins220 covers such diverse functions as the control of neuronal survival pathways, neurite outgrowth and maturation, and finally neuronal activity. Cumulative evidence from several studies supported the idea that neuronal activity in the hippocampus is reciprocally connected to Kidins220 protein levels, which appear to affect excitatory and inhibitory circuits in opposite ways (
      • Cortés R.Y.
      • Arévalo J.C.
      • Magby J.P.
      • Chao M.V.
      • Plummer M.R.
      Developmental and activity-dependent regulation of ARMS/Kidins220 in cultured rat hippocampal neurons.
      ,
      • Arévalo J.C.
      • Wu S.H.
      • Takahashi T.
      • Zhang H.
      • Yu T.
      • Yano H.
      • Milner T.A.
      • Tessarollo L.
      • Ninan I.
      • Arancio O.
      • Chao M.V.
      The ARMS/Kidins220 scaffold protein modulates synaptic transmission.
      ,
      • Wu S.H.
      • Arévalo J.C.
      • Neubrand V.E.
      • Zhang H.
      • Arancio O.
      • Chao M.V.
      The ankyrin repeat-rich membrane spanning (ARMS)/Kidins220 scaffold protein is regulated by activity-dependent calpain proteolysis and modulates synaptic plasticity.
      ,
      • Sutachan J.J.
      • Chao M.V.
      • Ninan I.
      Regulation of inhibitory neurotransmission by the scaffolding protein ankyrin repeat-rich membrane spanning/kinase D-interacting substrate of 220 kDa.
      ). This connection holds true also in the reverse direction, as sustained neuronal activity reduces the amount of Kidins220 protein via transcriptional down-regulation and calpain-dependent protein cleavage (
      • Wu S.H.
      • Arévalo J.C.
      • Neubrand V.E.
      • Zhang H.
      • Arancio O.
      • Chao M.V.
      The ankyrin repeat-rich membrane spanning (ARMS)/Kidins220 scaffold protein is regulated by activity-dependent calpain proteolysis and modulates synaptic plasticity.
      ,
      • López-Menéndez C.
      • Gascón S.
      • Sobrado M.
      • Vidaurre O.G.
      • Higuero A.M.
      • Rodríguez-Peña A.
      • Iglesias T.
      • Díaz-Guerra M.
      Kidins220/ARMS downregulation by excitotoxic activation of NMDARs reveals its involvement in neuronal survival and death pathways.
      ). The molecular mechanisms by which Kidins220 affects neuronal activity are only partially understood. In some instances, similarly to other multidomain scaffold proteins, such as PSD-95 (
      • Shin H.
      • Hsueh Y.P.
      • Yang F.C.
      • Kim E.
      • Sheng M.
      An intramolecular interaction between Src homology 3 domain and guanylate kinase-like domain required for channel clustering by postsynaptic density-95/SAP90.
      ) or zonula occludens (
      • Hervé J.C.
      • Derangeon M.
      • Sarrouilhe D.
      • Bourmeyster N.
      Influence of the scaffolding protein zonula occludens (ZOs) on membrane channels.
      ), an association of Kidins220 with membrane channels appears to be involved, as demonstrated by immunoprecipitation assays for AMPA and NMDA receptors (
      • Arévalo J.C.
      • Wu S.H.
      • Takahashi T.
      • Zhang H.
      • Yu T.
      • Yano H.
      • Milner T.A.
      • Tessarollo L.
      • Ninan I.
      • Arancio O.
      • Chao M.V.
      The ARMS/Kidins220 scaffold protein modulates synaptic transmission.
      ,
      • López-Menéndez C.
      • Gascón S.
      • Sobrado M.
      • Vidaurre O.G.
      • Higuero A.M.
      • Rodríguez-Peña A.
      • Iglesias T.
      • Díaz-Guerra M.
      Kidins220/ARMS downregulation by excitotoxic activation of NMDARs reveals its involvement in neuronal survival and death pathways.
      ). In the case of AMPA receptors, Kidins220 expression negatively regulated the basal synaptic strength of glutamatergic hippocampal synapses by affecting the phosphorylation state and cell surface expression of the GluA1 subunit (
      • Arévalo J.C.
      • Wu S.H.
      • Takahashi T.
      • Zhang H.
      • Yu T.
      • Yano H.
      • Milner T.A.
      • Tessarollo L.
      • Ninan I.
      • Arancio O.
      • Chao M.V.
      The ARMS/Kidins220 scaffold protein modulates synaptic transmission.
      ). In other instances, the effects on neuronal activity are most likely related to the close association of Kidins220 with neurotrophin receptors. In fact, Kidins220 has been shown to interact with the three members of the Trk family and with the p75 neurotrophin receptor (
      • Arévalo J.C.
      • Yano H.
      • Teng K.K.
      • Chao M.V.
      A unique pathway for sustained neurotrophin signaling through an ankyrin-rich membrane-spanning protein.
      ,
      • Chang M.S.
      • Arevalo J.C.
      • Chao M.V.
      Ternary complex with Trk, p75, and an ankyrin-rich membrane spanning protein.
      ). Moreover, Kidins220 is tyrosine phosphorylated by Trk receptors upon activation by extracellular neurotrophins (
      • Kong H.
      • Boulter J.
      • Weber J.L.
      • Lai C.
      • Chao M.V.
      An evolutionarily conserved transmembrane protein that is a novel downstream target of neurotrophin and ephrin receptors.
      ). Neurotrophins, and in particular brain-derived neurotrophic factor (BDNF), are known to participate in synaptic plasticity (
      • Poo M.M.
      Neurotrophins as synaptic modulators.
      ), and in some cases an involvement of Kidins220 in the synaptic effects of neurotrophins has been demonstrated. Kidins220 knock-down in hippocampal neurons interfered with the BDNF-evoked enhancement of inhibitory neurotransmission (
      • Sutachan J.J.
      • Chao M.V.
      • Ninan I.
      Regulation of inhibitory neurotransmission by the scaffolding protein ankyrin repeat-rich membrane spanning/kinase D-interacting substrate of 220 kDa.
      ). Long-term potentiation in mouse hippocampal slices, for which a direct implication of the BDNF-TrkB system has been shown (
      • Minichiello L.
      TrkB signalling pathways in LTP and learning.
      ), was enhanced in Kidins220+/− mice having reduced protein levels (
      • Wu S.H.
      • Arévalo J.C.
      • Neubrand V.E.
      • Zhang H.
      • Arancio O.
      • Chao M.V.
      The ankyrin repeat-rich membrane spanning (ARMS)/Kidins220 scaffold protein is regulated by activity-dependent calpain proteolysis and modulates synaptic plasticity.
      ). Finally, the potentiation of excitatory post-synaptic currents in hippocampal neurons induced by acute BDNF application (
      • Levine E.S.
      • Dreyfus C.F.
      • Black I.B.
      • Plummer M.R.
      Brain-derived neurotrophic factor rapidly enhances synaptic transmission in hippocampal neurons via postsynaptic tyrosine kinase receptors.
      ) was impaired in Kidins220−/− mice (
      • Cesca F.
      • Yabe A.
      • Spencer-Dene B.
      • Scholz-Starke J.
      • Medrihan L.
      • Maden C.H.
      • Gerhardt H.
      • Orriss I.R.
      • Baldelli P.
      • Al-Qatari M.
      • Koltzenburg M.
      • Adams R.H.
      • Benfenati F.
      • Schiavo G.
      Kidins220/ARMS mediates the integration of the neurotrophin and VEGF pathways in the vascular and nervous systems.
      ).
      The proper functioning of brain circuits relies on the ability of the neural networks to maintain the correct balance between excitatory and inhibitory activity. Neuronal excitability is determined by a complex network of biological processes, which result from the interplay of extracellular signals, membrane receptors and channels, and intracellular signaling cascades. Being responsible for AP onset at the axonal initial segment, Nav channels are fundamental players in all kinds of neuronal communication. They are multimeric complexes of α and β subunits that exist in several isoforms, creating a large repertoire of channels with different biophysical properties (
      • Lai H.C.
      • Jan L.Y.
      The distribution and targeting of neuronal voltage-gated ion channels.
      ). The specific channel localization is determined by the interaction of α and β subunits with a set of adhesion molecules, as well as cytoskeletal and extracellular matrix proteins (
      • Lai H.C.
      • Jan L.Y.
      The distribution and targeting of neuronal voltage-gated ion channels.
      ,
      • Bouzidi M.
      • Tricaud N.
      • Giraud P.
      • Kordeli E.
      • Caillol G.
      • Deleuze C.
      • Couraud F.
      • Alcaraz G.
      Interaction of the Nav1.2 α subunit of the voltage-dependent sodium channel with nodal ankyrinG: in vitro mapping of the interacting domains and association in synaptosomes.
      ). Nav channel activity is modulated by the coordinated activation of several signaling pathways, being targets of phosphorylation by protein kinase A (
      • Li M.
      • West J.W.
      • Lai Y.
      • Scheuer T.
      • Catterall W.A.
      Functional modulation of brain sodium channels by cAMP-dependent phosphorylation.
      ) and by Fyn kinase acting downstream of the BDNF-TrkB complex (
      • Ahn M.
      • Beacham D.
      • Westenbroek R.E.
      • Scheuer T.
      • Catterall W.A.
      Regulation of Nav1.2 channels by brain-derived neurotrophic factor, TrkB, and associated Fyn kinase.
      ). Despite the vast amount of data available in the literature, however, the network of signaling events controlling Nav channel localization and activity is still far from being completely understood. Here, we examined the effects of Kidins220 ablation on the excitability of cultured hippocampal neurons. Our results revealed increased excitability of GABAergic neurons in Kidins220 null mice and provided evidence for a functional association of Kidins220 with Nav channels in the brain.

      Discussion

      In the present study, we investigated the role of the scaffold protein Kidins220 in the regulation of neuronal excitability using embryonal hippocampal neurons isolated from the previously described Kidins220−/− full knock-out mice (
      • Cesca F.
      • Yabe A.
      • Spencer-Dene B.
      • Scholz-Starke J.
      • Medrihan L.
      • Maden C.H.
      • Gerhardt H.
      • Orriss I.R.
      • Baldelli P.
      • Al-Qatari M.
      • Koltzenburg M.
      • Adams R.H.
      • Benfenati F.
      • Schiavo G.
      Kidins220/ARMS mediates the integration of the neurotrophin and VEGF pathways in the vascular and nervous systems.
      ). Current-clamp recordings revealed increased excitability in GABAergic and, to a lower extent, in glutamatergic Kidins220−/− neurons. We observed higher action potential rising slopes and peak amplitudes in Kidins220−/− GABAergic neurons, pointing to an increased Na+ conductance, which lead us to deduce a misregulation of Nav channels in these cells. Accordingly, we conducted co-immunoprecipitation experiments on mouse cortical brain lysates and HEK293 cells transiently expressing Nav1.2, which supported the association between Kidins220 and α subunits of neuronal Nav channels. Importantly, whole cell patch clamp experiments showed that Kidins220 co-transfection in HEK293 cells caused complex changes in Nav1.2 channel properties, which were partly consistent with decreased excitability (positive shift of the activation curve, slower activation kinetics), partly with increased excitability (positive shift of the steady-state inactivation curve, slower inactivation kinetics, and faster recovery from inactivation). The modifications exerted by Kidins220 on Nav1.2 channels are new and substantial. They indicated that two separate processes of Nav channel function were affected: (i) gate opening as a result of the movement of the voltage-sensing transmembrane domains (channel activation) (
      • Catterall W.A.
      Ion channel voltage sensors: structure, function, and pathophysiology.
      ), and (ii) subsequent pore occlusion by an inactivating blocking particle through a “hinged-lid” mechanism (fast channel inactivation) (
      • Goldin A.L.
      Mechanisms of sodium channel inactivation.
      ).
      Kidins220 association with brain Nav channels gains further importance in view of the well established connection between BDNF signaling and neuronal excitability. In fact, being a direct target of Trk neurotrophin receptors, the Kidins220 protein could be in a strategic intermediary position between TrkB receptor activation and downstream ion channel modulation. Notably, among the different examples present in the literature (
      • Li H.S.
      • Xu X.Z.
      • Montell C.
      Activation of a TRPC3-dependent cation current through the neurotrophin BDNF.
      ,
      • Rogalski S.L.
      • Appleyard S.M.
      • Pattillo A.
      • Terman G.W.
      • Chavkin C.
      TrkB activation by brain-derived neurotrophic factor inhibits the G protein-gated inward rectifier Kir3 by tyrosine phosphorylation of the channel.
      ,
      • Tucker K.
      • Fadool D.A.
      Neurotrophin modulation of voltage-gated potassium channels in rat through TrkB receptors is time and sensory experience dependent.
      ,
      • Nieto-Gonzalez J.L.
      • Jensen K.
      BDNF depresses excitability of parvalbumin-positive interneurons through an M-like current in rat dentate gyrus.
      ), some specifically concern Nav channels: Nav1.2 channels are subject to dynamic regulation by tyrosine phosphorylation/dephosphorylation events, with Fyn kinase acting to increase fast inactivation in response to TrkB activation (
      • Ahn M.
      • Beacham D.
      • Westenbroek R.E.
      • Scheuer T.
      • Catterall W.A.
      Regulation of Nav1.2 channels by brain-derived neurotrophic factor, TrkB, and associated Fyn kinase.
      ,
      • Beacham D.
      • Ahn M.
      • Catterall W.A.
      • Scheuer T.
      Sites and molecular mechanisms of modulation of Nav1.2 channels by Fyn tyrosine kinase.
      ) and the tyrosine phosphatase RPTPβ having the opposite effect (
      • Ratcliffe C.F.
      • Qu Y.
      • McCormick K.A.
      • Tibbs V.C.
      • Dixon J.E.
      • Scheuer T.
      • Catterall W.A.
      A sodium channel signaling complex: modulation by associated receptor protein tyrosine phosphatase β.
      ). Furthermore, BDNF (in complex with TrkB receptors) has been proposed to elicit the rapid activation of Nav1.9 channels, which appears to occur in a ligand-gated rather than in a voltage-gated manner (
      • Blum R.
      • Kafitz K.W.
      • Konnerth A.
      Neurotrophin-evoked depolarization requires the sodium channel NaV1.9.
      ). BDNF-dependent Nav1.9 channel activation lead to fast membrane depolarization and increased excitability in different neuronal cell types (
      • Kafitz K.W.
      • Rose C.R.
      • Thoenen H.
      • Konnerth A.
      Neurotrophin-evoked rapid excitation through TrkB receptors.
      ). The modulation of Nav1.2 channel properties in HEK293 cells reported here depended solely on Kidins220 co-expression. Kidins220 may convey extracellular signals to the channels, via phosphorylation of specific sites or by assembling a platform of adaptor/signaling proteins around the channels to initiate the appropriate intracellular response. Given the prominent role of Kidins220 as a mediator of neurotrophin signaling, it will be important to study whether the Kidins220-Nav channel interaction is modulated by the TrkB/BDNF signaling pathway.
      Clearly, the effects of Kidins220 expression on neuronal excitability are not easily predictable from our co-expression data, principally because of the complexity of Nav1.2 channel modification and also because neurons express diverse isoforms of Nav α subunits (among which also Nav1.2) at different densities in distinct neuronal compartments. Kidins220 ablation apparently increased the Na+ conductance in GABAergic hippocampal neurons, which may be due to higher plasma membrane expression of Nav α subunits or to shifts in the voltage dependence of Nav channel activation or fast inactivation. Our data suggest that Kidins220 modulates neuronal excitability by acting, at least partly, on Nav channel properties, which is supported by simulation data showing that introducing the Kidins220-dependent Na+ current alterations in an in silico model of CA1/CA3 hippocampal interneurons was sufficient to recapitulate the main findings observed in Kidins220−/− GABAergic neurons. However, the involvement of alternative mechanisms cannot be excluded. The findings presented here raise a number of interesting questions concerning the α subunit specificity, the subcellular localization, and cell-type specificity of Kidins220 association, which will be a subject of future investigation.
      Notably, increased excitability of Kidins220−/− neurons manifested itself principally in inhibitory neurons, similarly to what has been observed for synaptic plasticity paradigms (
      • Scholz-Starke J.
      • Cesca F.
      • Schiavo G.
      • Benfenati F.
      • Baldelli P.
      Kidins220/ARMS is a novel modulator of short-term synaptic plasticity in hippocampal GABAergic neurons.
      ). The reason for this is currently unclear, but is likely to be related to cell-specific differences in the expression and/or subcellular localization of Kidins220-interacting proteins, as Kidins220 localizes to soma and processes of both inhibitory and excitatory neurons (
      • Sutachan J.J.
      • Chao M.V.
      • Ninan I.
      Regulation of inhibitory neurotransmission by the scaffolding protein ankyrin repeat-rich membrane spanning/kinase D-interacting substrate of 220 kDa.
      ,
      • Scholz-Starke J.
      • Cesca F.
      • Schiavo G.
      • Benfenati F.
      • Baldelli P.
      Kidins220/ARMS is a novel modulator of short-term synaptic plasticity in hippocampal GABAergic neurons.
      ). In both cases, i.e. synaptic plasticity and intrinsic excitability of inhibitory neurons, Kidins220 ablation appeared to cause gain-of-function phenotypes: inhibitory post-synaptic currents showed faster recovery from synaptic depression in response to paired-pulse and prolonged train stimulation (
      • Scholz-Starke J.
      • Cesca F.
      • Schiavo G.
      • Benfenati F.
      • Baldelli P.
      Kidins220/ARMS is a novel modulator of short-term synaptic plasticity in hippocampal GABAergic neurons.
      ); here we showed that GABAergic neurons required less current injection to elicit AP firing and reached higher spiking frequencies. Interestingly, blockade of endogenous TrkB receptors in pyramidal neurons from rat visual cortex similarly reduced the threshold current and increased the instantaneous firing frequency (
      • Desai N.S.
      • Rutherford L.C.
      • Turrigiano G.G.
      BDNF regulates the intrinsic excitability of cortical neurons.
      ). Both Kidins220−/− phenotypes would be expected to reinforce the weight of synaptic inhibition within the neuronal network, thereby reducing its overall activity. MEA recordings on hippocampal neurons fully confirmed this prediction, as they revealed a striking impairment of delayed spiking activity in Kidins220−/− cultures subjected to pulse stimulation.
      The reduction of the threshold current in Kidins220−/− GABAergic neurons was likely due to specific membrane conductance changes in the subthreshold potential range, because resting membrane potential and input resistance were unaffected. Apart from Nav channels, Kidins220 ablation may also cause alterations of M-type K+ currents mediated by Kv7/KCNQ channels, known to be involved in the regulation of interneuronal excitability (
      • Lawrence J.J.
      • Saraga F.
      • Churchill J.F.
      • Statland J.M.
      • Travis K.E.
      • Skinner F.K.
      • McBain C.J.
      Somatodendritic Kv7/KCNQ/M channels control interspike interval in hippocampal interneurons.
      ) and to be modulated by a variety of neurotransmitters and hormones (
      • Brown D.A.
      • Passmore G.M.
      Neural KCNQ (Kv7) channels.
      ). This hypothesis gains in weight by recent results showing a potentiation of M-type K+ currents in parvalbumin-positive hippocampal interneurons by BDNF (
      • Nieto-Gonzalez J.L.
      • Jensen K.
      BDNF depresses excitability of parvalbumin-positive interneurons through an M-like current in rat dentate gyrus.
      ).
      Altogether, our results identify Kidins220 as a powerful new modulator of Nav channel function, indicating a potential role in the control of neuronal excitability, both at the single-cell level and the network level. Interestingly, Kidins220 action seems to be more evident in GABAergic neurons, suggesting that it may participate in the maintenance of the balance between excitation and inhibition in neural networks. These results deepen our understanding of the molecular mechanisms modulating neural excitability and network activity and may open new perspectives to target the increasing number of pathologies associated with a dysregulation of Nav channel function and altered network excitability.

      Author Contributions

      J. S. S., F. C., and F. B. designed the study. J. S. S. (patch clamp electrophysiology), E. F. (MEA electrophysiology), A. S. and F. C. (biochemistry) performed the experiments and analyzed the data. T. N. performed modeling work. J. S. S. and F. C. wrote the manuscript. All authors reviewed the results and approved the final version of the manuscript.

      Acknowledgments

      We thank Cristiana Picco, Anna Boccaccio (IBF-CNR, Genoa) and Fanny Jaudon (IIT, Genoa) for valuable advice, useful discussions and critical reading of the manuscript, Pietro Baldelli (University of Genoa) for support in early stages of the study, and Marina Nanni (IIT) for excellent technical assistance. We are grateful to G. P. Schiavo (London, UK) and W. A. Catterall (Seattle, Washington) for providing the Kidins220 and Nav1.2 constructs.

      References

        • Iglesias T.
        • Cabrera-Poch N.
        • Mitchell M.P.
        • Naven T.J.
        • Rozengurt E.
        • Schiavo G.
        Identification and cloning of Kidins220, a novel neuronal substrate of protein kinase D.
        J. Biol. Chem. 2000; 275: 40048-40056
        • Kong H.
        • Boulter J.
        • Weber J.L.
        • Lai C.
        • Chao M.V.
        An evolutionarily conserved transmembrane protein that is a novel downstream target of neurotrophin and ephrin receptors.
        J. Neurosci. 2001; 21: 176-185
        • Neubrand V.E.
        • Cesca F.
        • Benfenati F.
        • Schiavo G.
        Kidins220/ARMS as a functional mediator of multiple receptor signalling pathways.
        J. Cell Sci. 2012; 125: 1845-1854
        • Cortés R.Y.
        • Arévalo J.C.
        • Magby J.P.
        • Chao M.V.
        • Plummer M.R.
        Developmental and activity-dependent regulation of ARMS/Kidins220 in cultured rat hippocampal neurons.
        Dev. Neurobiol. 2007; 67: 1687-1698
        • Arévalo J.C.
        • Wu S.H.
        • Takahashi T.
        • Zhang H.
        • Yu T.
        • Yano H.
        • Milner T.A.
        • Tessarollo L.
        • Ninan I.
        • Arancio O.
        • Chao M.V.
        The ARMS/Kidins220 scaffold protein modulates synaptic transmission.
        Mol. Cell. Neurosci. 2010; 45: 92-100
        • Wu S.H.
        • Arévalo J.C.
        • Neubrand V.E.
        • Zhang H.
        • Arancio O.
        • Chao M.V.
        The ankyrin repeat-rich membrane spanning (ARMS)/Kidins220 scaffold protein is regulated by activity-dependent calpain proteolysis and modulates synaptic plasticity.
        J. Biol. Chem. 2010; 285: 40472-40478
        • Sutachan J.J.
        • Chao M.V.
        • Ninan I.
        Regulation of inhibitory neurotransmission by the scaffolding protein ankyrin repeat-rich membrane spanning/kinase D-interacting substrate of 220 kDa.
        J. Neurosci. Res. 2010; 88: 3447-3456
        • López-Menéndez C.
        • Gascón S.
        • Sobrado M.
        • Vidaurre O.G.
        • Higuero A.M.
        • Rodríguez-Peña A.
        • Iglesias T.
        • Díaz-Guerra M.
        Kidins220/ARMS downregulation by excitotoxic activation of NMDARs reveals its involvement in neuronal survival and death pathways.
        J. Cell Sci. 2009; 122: 3554-3565
        • Shin H.
        • Hsueh Y.P.
        • Yang F.C.
        • Kim E.
        • Sheng M.
        An intramolecular interaction between Src homology 3 domain and guanylate kinase-like domain required for channel clustering by postsynaptic density-95/SAP90.
        J. Neurosci. 2000; 20: 3580-3587
        • Hervé J.C.
        • Derangeon M.
        • Sarrouilhe D.
        • Bourmeyster N.
        Influence of the scaffolding protein zonula occludens (ZOs) on membrane channels.
        Biochim. Biophys. Acta. 2014; 1838: 595-604
        • Arévalo J.C.
        • Yano H.
        • Teng K.K.
        • Chao M.V.
        A unique pathway for sustained neurotrophin signaling through an ankyrin-rich membrane-spanning protein.
        EMBO J. 2004; 23: 2358-2368
        • Chang M.S.
        • Arevalo J.C.
        • Chao M.V.
        Ternary complex with Trk, p75, and an ankyrin-rich membrane spanning protein.
        J. Neurosci. Res. 2004; 78: 186-192
        • Poo M.M.
        Neurotrophins as synaptic modulators.
        Nat. Rev. Neurosci. 2001; 2: 24-32
        • Minichiello L.
        TrkB signalling pathways in LTP and learning.
        Nat. Rev. Neurosci. 2009; 10: 850-860
        • Levine E.S.
        • Dreyfus C.F.
        • Black I.B.
        • Plummer M.R.
        Brain-derived neurotrophic factor rapidly enhances synaptic transmission in hippocampal neurons via postsynaptic tyrosine kinase receptors.
        Proc. Natl. Acad. Sci. U.S.A. 1995; 92: 8074-8077
        • Cesca F.
        • Yabe A.
        • Spencer-Dene B.
        • Scholz-Starke J.
        • Medrihan L.
        • Maden C.H.
        • Gerhardt H.
        • Orriss I.R.
        • Baldelli P.
        • Al-Qatari M.
        • Koltzenburg M.
        • Adams R.H.
        • Benfenati F.
        • Schiavo G.
        Kidins220/ARMS mediates the integration of the neurotrophin and VEGF pathways in the vascular and nervous systems.
        Cell Death Differ. 2012; 19: 194-208
        • Lai H.C.
        • Jan L.Y.
        The distribution and targeting of neuronal voltage-gated ion channels.
        Nat. Rev. Neurosci. 2006; 7: 548-562
        • Bouzidi M.
        • Tricaud N.
        • Giraud P.
        • Kordeli E.
        • Caillol G.
        • Deleuze C.
        • Couraud F.
        • Alcaraz G.
        Interaction of the Nav1.2 α subunit of the voltage-dependent sodium channel with nodal ankyrinG: in vitro mapping of the interacting domains and association in synaptosomes.
        J. Biol. Chem. 2002; 277: 28996-29004
        • Li M.
        • West J.W.
        • Lai Y.
        • Scheuer T.
        • Catterall W.A.
        Functional modulation of brain sodium channels by cAMP-dependent phosphorylation.
        Neuron. 1992; 8: 1151-1159
        • Ahn M.
        • Beacham D.
        • Westenbroek R.E.
        • Scheuer T.
        • Catterall W.A.
        Regulation of Nav1.2 channels by brain-derived neurotrophic factor, TrkB, and associated Fyn kinase.
        J. Neurosci. 2007; 27: 11533-11542
        • Bekkers J.M.
        • Stevens C.F.
        Excitatory and inhibitory autaptic currents in isolated hippocampal neurons maintained in cell culture.
        Proc. Natl. Acad. Sci. U.S.A. 1991; 88: 7834-7838
        • Scholz-Starke J.
        • Cesca F.
        • Schiavo G.
        • Benfenati F.
        • Baldelli P.
        Kidins220/ARMS is a novel modulator of short-term synaptic plasticity in hippocampal GABAergic neurons.
        PLoS One. 2012; 7e35785
        • Bologna L.L.
        • Pasquale V.
        • Garofalo M.
        • Gandolfo M.
        • Baljon P.L.
        • Maccione A.
        • Martinoia S.
        • Chiappalone M.
        Investigating neuronal activity by SPYCODE multi-channel data analyzer.
        Neural Netw. 2010; 23: 685-697
        • Minneci F.
        • Janahmadi M.
        • Migliore M.
        • Dragicevic N.
        • Avossa D.
        • Cherubini E.
        Signaling properties of stratum oriens interneurons in the hippocampus of transgenic mice expressing EGFP in a subset of somatostatin-containing cells.
        Hippocampus. 2007; 17: 538-553
        • Bean B.P.
        The action potential in mammalian central neurons.
        Nat. Rev. Neurosci. 2007; 8: 451-465
        • Debanne D.
        • Campanac E.
        • Bialowas A.
        • Carlier E.
        • Alcaraz G.
        Axon physiology.
        Physiol. Rev. 2011; 91: 555-602
        • Mantegazza M.
        • Catterall W.A.
        Voltage-gated Na+ channels: structure, function, and pathophysiology.
        in: Noebels J.L. Avoli M. Rogawski M.A. Olsen R.W. Delgado-Escueta A.V. Jasper's Basic Mechanisms of the Epilepsies (Internet). 4th Ed. National Center for Biotechnology Information, Bethesda, MD2012
        • Li T.
        • Tian C.
        • Scalmani P.
        • Frassoni C.
        • Mantegazza M.
        • Wang Y.
        • Yang M.
        • Wu S.
        • Shu Y.
        Action potential initiation in neocortical inhibitory interneurons.
        PLoS Biol. 2014; 12e1001944
        • Catterall W.A.
        Ion channel voltage sensors: structure, function, and pathophysiology.
        Neuron. 2010; 67: 915-928
        • Goldin A.L.
        Mechanisms of sodium channel inactivation.
        Curr. Opin. Neurobiol. 2003; 13: 284-290
        • Li H.S.
        • Xu X.Z.
        • Montell C.
        Activation of a TRPC3-dependent cation current through the neurotrophin BDNF.
        Neuron. 1999; 24: 261-273
        • Rogalski S.L.
        • Appleyard S.M.
        • Pattillo A.
        • Terman G.W.
        • Chavkin C.
        TrkB activation by brain-derived neurotrophic factor inhibits the G protein-gated inward rectifier Kir3 by tyrosine phosphorylation of the channel.
        J. Biol. Chem. 2000; 275: 25082-25088
        • Tucker K.
        • Fadool D.A.
        Neurotrophin modulation of voltage-gated potassium channels in rat through TrkB receptors is time and sensory experience dependent.
        J. Physiol. 2002; 542: 413-429
        • Nieto-Gonzalez J.L.
        • Jensen K.
        BDNF depresses excitability of parvalbumin-positive interneurons through an M-like current in rat dentate gyrus.
        PLoS One. 2013; 8e67318
        • Beacham D.
        • Ahn M.
        • Catterall W.A.
        • Scheuer T.
        Sites and molecular mechanisms of modulation of Nav1.2 channels by Fyn tyrosine kinase.
        J. Neurosci. 2007; 27: 11543-11551
        • Ratcliffe C.F.
        • Qu Y.
        • McCormick K.A.
        • Tibbs V.C.
        • Dixon J.E.
        • Scheuer T.
        • Catterall W.A.
        A sodium channel signaling complex: modulation by associated receptor protein tyrosine phosphatase β.
        Nat. Neurosci. 2000; 3: 437-444
        • Blum R.
        • Kafitz K.W.
        • Konnerth A.
        Neurotrophin-evoked depolarization requires the sodium channel NaV1.9.
        Nature. 2002; 419: 687-693
        • Kafitz K.W.
        • Rose C.R.
        • Thoenen H.
        • Konnerth A.
        Neurotrophin-evoked rapid excitation through TrkB receptors.
        Nature. 1999; 401: 918-921
        • Desai N.S.
        • Rutherford L.C.
        • Turrigiano G.G.
        BDNF regulates the intrinsic excitability of cortical neurons.
        Learn Mem. 1999; 6: 284-291
        • Lawrence J.J.
        • Saraga F.
        • Churchill J.F.
        • Statland J.M.
        • Travis K.E.
        • Skinner F.K.
        • McBain C.J.
        Somatodendritic Kv7/KCNQ/M channels control interspike interval in hippocampal interneurons.
        J. Neurosci. 2006; 26: 12325-12338
        • Brown D.A.
        • Passmore G.M.
        Neural KCNQ (Kv7) channels.
        Br. J. Pharmacol. 2009; 156: 1185-1195