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Impaired permeability to Ins(1,4,5)P3 in a mutant connexin underlies recessive hereditary deafness

Nature Cell Biology volume 7, pages 63–69 (2005)Cite this article

Abstract

Connexins are membrane proteins that assemble into gap-junction channels and are responsible for direct, electrical and metabolic coupling between connected cells. Here we describe an investigation of the properties of a recombinantly expressed recessive mutant of connexin 26 (Cx26), the V84L mutant, associated with deafness. Unlike other Cx26 mutations, V84L affects neither intracellular sorting nor electrical coupling, but specifically reduces permeability to the Ca2+-mobilizing messenger inositol 1,4,5-trisphosphate (Ins(1,4,5)P3). Both the permeability to Lucifer Yellow and the unitary channel conductance of V84L-mutant channels are indistinguishable from those of the wild-type Cx26. Injection of Ins(1,4,5)P3 into supporting cells of the rat organ of Corti, which abundantly express Cx26, ensues in a regenerative wave of Ca2+ throughout the tissue. Blocking the gap junction communication abolishes wave propagation. We propose that the V84L mutation reduces metabolic coupling mediated by Ins(1,4,5)P3 to an extent sufficient to impair the propagation of Ca2+ waves and the formation of a functional syncytium. Our data provide the first demonstration of a specific defect of metabolic coupling and offer a mechanistic explanation for the pathogenesis of an inherited human disease.

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Figure 1: Electrical conductance and permeability to Lucifer Yellow are normal in homotypic gap-junction channels formed by the V84L mutant of Cx26.
Figure 2: Intercellular transfer of IP3 is impaired in V84L transfectants.
Figure 3: Measurement of Ins(1,4,5)P3 permeability in transfected pairs of HeLa cells.
Figure 4: Intercellular transfer of Ins(1,4,5)P3 in supporting cells of the organ of Corti.

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Acknowledgements

This work was supported by grants from the Telethon Foundation, (Project n. GGP02043), the Ministero dell' Università e Ricerca Scientifica (MIUR, FIRB n. RBAU01Z2Z8, PRIN-COFIN 2002067312_002) to F.M., the Centro di Eccellenza (co-ordinator, T.P.) and the Italian Health Ministry. We thank R. Bruzzone (Institute Pasteur, Paris, France) for helpful comments, S. Bastianello, M. Bortolozzi and C. D. Ciubotaru (Venetian Institute of Molecular Medicine, Padua, Italy) for help with computer programming and image processing.

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Author notes

  1. Martina Beltramello and Valeria Piazza: These authors contributed equally to this work.

Authors and Affiliations

  1. Venetian Institute of Molecular Medicine, via G. Orus 2, Padua, 35129, Italy

    Martina Beltramello, Valeria Piazza, Tullio Pozzan & Fabio Mammano

  2. Department of Neuroscience, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, 10461, New York, USA

    Feliksas F. Bukauskas

  3. Department of Biomedical Sciences and Consiglio Nazionale delle Ricerche Institute of Neuroscience, University of Padua, viale G. Colombo 3, Padua, 35121, Italy

    Tullio Pozzan

  4. Department of Physics, University, of Padua, via Marzolo 8, Padua, 35132, Italy

    Fabio Mammano

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  1. Martina Beltramello
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  3. Feliksas F. Bukauskas
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Correspondence to Fabio Mammano.

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Beltramello, M., Piazza, V., Bukauskas, F. et al. Impaired permeability to Ins(1,4,5)P3 in a mutant connexin underlies recessive hereditary deafness. Nat Cell Biol 7, 63–69 (2005). https://doi.org/10.1038/ncb1205

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