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Regulation of voltage-gated sodium current by endogenous Src family kinases in cochlear spiral ganglion neurons in culture

  • Ion Channels, Receptors and Transporters
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Abstract

Voltage-gated sodium (Na+) and potassium (K+) channels have been found to be regulated by Src family kinases (SFKs). However, how these channels are regulated by SFKs in cochlear spiral ganglion neurons (SGNs) remains unknown. Here, we report that altering the activity of endogenous SFKs modulated voltage-gated Na+, but not K+, currents recorded in embryonic SGNs in culture. Voltage-gated Na+ current was suppressed by inhibition of endogenous SFKs or just Src and potentiated by the activation of these enzymes. Detailed investigations showed that under basal conditions, SFK inhibitor application did not significantly affect the voltage-dependent activation, but shifted the steady-state inactivation curves of Na+ currents and delayed the recovery of Na+ currents from inactivation. Application of Src specific inhibitor, Src40–58, not only shifted the inactivation curve but also delayed the recovery of Na+ currents and moved the voltage-dependent activation curve towards the left. The pre-inhibition of SFKs occluded all the effects induced by Src40–58 application, except the left shift of the activation curve. The activation of SFKs did not change either steady-state inactivation or recovery of Na+ currents, but caused the left shift of the activation curve. SFK inhibitor application effectively prevented all the effects induced by SFK activation, suggesting that both the voltage-dependent activation and steady-state inactivation of Na+ current are subjects of SFK regulation. The different effects induced by activation versus inhibition of SFKs implied that under basal conditions, endogenously active and inactive SFKs might be differentially involved in the regulation of voltage-gated Na+ channels in SGNs.

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Acknowledgements

SF is supported by the State Scholarship Fund of China (2009845013). This work was supported by a grant from NIH (5R01 NS053567-04) to XMY. We would like to thank Dr. M. W. Salter for kindly supplying us Src inhibitory peptide (Src40–58), SFK activator peptide (EPQ(pY)EEIPIA), and control peptide (EPQYEEIPIA). We would also like to thank Drs. M. W. Salter, M. Hildebrand, and G. Pitcher for the very constructive discussions.

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Authors and Affiliations

  1. Department of Otolaryngology—Head and Neck Surgery, First Affiliated Hospital, Guangxi Medical University, Nanning, Guangxi, 530021, People’s Republic of China

    Shuang Feng, Jiping Su & Xian-Min Yu

  2. Department of Biomedical Sciences, Florida State University, Tallahassee, FL, 32306, USA

    Shuang Feng, Melissa Pflueger, Shuang-Xiu Lin, Bradley R. Groveman & Xian-Min Yu

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  1. Shuang Feng
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Correspondence to Jiping Su or Xian-Min Yu.

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Supplementary Fig. 1

Expression of SFKs in SGNs. N-200: an antibody staining for neurofilament 200; SFKs: an antibody staining for SFKs. Merge: merged image showing co-labeling of neurofilament 200 and SFKs (TIFF 716 KB)

Supplementary Fig. 2

Effects of intracellular application of the SFK activator peptide EPQ(pY)EEIPIA (1 mM) on TTX-resistant inward current. A: example of current traces recorded immediately (0’) and 5 min after breakthrough with electrodes filled with intracellular solution containing the SFK activator peptide, EPQ(pY)EEIPIA (1 mM) from a SGN bathed with extracellular solution containing TTX (1 μM). B: Summary data showing relative changes in TTX-resistant currents recorded from SGNs applied intracellularly with the SFK activator peptide. Values in brackets indicate number of SGNs tested (TIFF 9.10 MB)

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Feng, S., Pflueger, M., Lin, SX. et al. Regulation of voltage-gated sodium current by endogenous Src family kinases in cochlear spiral ganglion neurons in culture. Pflugers Arch - Eur J Physiol 463, 571–584 (2012). https://doi.org/10.1007/s00424-012-1072-4

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