Actin-independent trafficking of cochlear connexin 26 to non-lipid raft gap junction plaques.

[en] Hereditary hearing loss affects about 1 per 1000 children. Mutations in GJB2, which encodes the connexin 26 protein (Cx26) involved in cochlear homeostasis, are found in about 50% of patients with autosomal recessive non-syndromic hearing loss. Deciphering the trafficking pathway of cochlear Cx26 in situ should represent an advance in understanding the pathogenic significance of many of these mutations. Connexins trafficking and delivery to lipid raft-associated gap junction plaques usually requires successively microtubule and actin networks. Here we show that cochlear Cx26 exhibits an unusual trafficking pathway. We observed that Cx26 assembly occurs in non-lipid raft membrane domains and that junctional plaques are devoid of actin and associated zonula occludens proteins. Using cytoskeleton-disrupting drugs in organotypic culture, we found that cochlear Cx26 gap junction assembly requires microtubules but not actin filaments. Altogether, our data provide an unexpected insight into Cx26 trafficking pathway and gap junction assembly in the cochlea.

Disciplines :

Biochemistry, biophysics & molecular biology

Author, co-author :

Defourny, Jean ; Université de Liège - ULiège > Neurosciences-Cellular and tissular biology

Thelen, Nicolas ; Université de Liège - ULiège > Département des sciences de la vie > Biologie cellulaire

Thiry, Marc ; Université de Liège - ULiège > Département des sciences de la vie > Biologie cellulaire

Language :

English

Title :

Actin-independent trafficking of cochlear connexin 26 to non-lipid raft gap junction plaques.

Publication date :

2019

Journal title :

Hearing Research

ISSN :

0378-5955

eISSN :

1878-5891

Publisher :

Elsevier, Netherlands

Peer reviewed :

Peer Reviewed verified by ORBi

Commentary :

Available on ORBi :

since 09 February 2019

Statistics

Number of views

141 (17 by ULiège)

Number of downloads

223 (10 by ULiège)

More statistics

Scopus citations^®

Scopus citations^®
without self-citations

OpenCitations

Bibliography

Ahmad, S., Chen, S., Sun, J., Lin, X., Connexins 26 and 30 are co-assembled to form gap junctions in the cochlea of mice. Biochem. Biophys. Res. Commun. 307 (2003), 362–368 https://doi.org/10.1016/S0006-291X(03)01166-5.
Ahmad, S., Evans, W.H., Post-translational integration and oligomerization of connexin 26 in plasma membranes and evidence of formation of membrane pores: implications for the assembly of gap junctions. Biochem. J. 365 (2002), 603–699 https://doi.org/10.1042/bj20011572.
Ambrosi, C., Walker, A.E., Depriest, A.D., Cone, A.C., Lu, C., Badger, J., Skerrett, I.M., Sosinsky, G.E., Analysis of trafficking, stability and function of human connexin 26 gap junction channels with deafness-causing mutations in the fourth transmembrane helix. PLoS One, 8, 2013, e70916 https://doi.org/10.1371/journal.pone.0070916.
Avshalumova, L., Fabrikant, J., Koriakos, A., Overview of skin diseases linked to connexin gene mutations. Int. J. Dermatol. 53 (2014), 192–205 https://doi.org/10.1111/ijd.12062.
Cascio, M., Connexins and their environment: effects of lipids composition on ion channels. Biochim. Biophys. Acta 1711 (2005), 142–153 https://doi.org/10.1016/j.bbamem.2004.12.001.
Chang, Q., Tang, W., Ahmad, S., Zhou, B., Lin, X., Gap junction mediated intercellular metabolite transfer in the cochlea is compromised in connexin30 null mice. PLoS One, 3, 2008, e4088 https://doi.org/10.1371/journal.pone.0004088.
Chichili, G.R., Rodgers, W., Cytoskeleton-membrane interactions in membrane raft structure. Cell. Mol. Life Sci. 66 (2009), 2319–2328 https://doi.org/10.1007/s00018-009-0022-6.
Davy, A., Bush, J.O., Soriano, P., Inhibition of gap junction communication at ectopic Eph/ephrin boundaries underlies craniofrontonasal syndrome. PLoS Biol., 4, 2006, e315 https://doi.org/10.1371/journal.pbio.0040315.
Defamie, N., Mesnil, M., The modulation of gap-junctional intercellular communication by lipid rafts. Biochim. Biophys. Acta 1818 (2012), 1866–1869 https://doi.org/10.1016/j.bbamem.2011.09.023.
Defourny, J., Mateo Sánchez, S., Schoonaert, L., Robberecht, W., Davy, A., Nguyen, L., Malgrange, B., Cochlear supporting cell transdifferentiation and integration into hair cell layers by inhibition of ephrin-B2 signalling. Nat. Commun., 6, 2015, 7017 https://doi.org/10.1038/ncomms8017.
Defourny, J., Thelen, N., Thiry, M., Cochlear connexin 30 homomeric and heteromeric channels exhibit distinct assembly mechanisms. Mech. Dev. 155 (2019), 8–14 https://doi.org/10.1016/j.mod.2018.10.001.
Diez, J.A., Ahmad, S., Evans, W.H., Assembly of heteromeric connexons in guinea-pig liver en route to the Golgi apparatus, plasma membrane and gap junctions. Eur. J. Biochem. 262 (1999), 142–148 https://doi.org/10.1046/j.1432-1327.1999.00343.x.
Ezratty, E.J., Partridge, M.A., Gundersen, G.G., Microtubule-induced focal adhesion disassembly is mediated by dynamin and focal adhesion kinase. Nat. Cell Biol. 7 (2005), 581–590 https://doi.org/10.1038/ncb1262.
Gaietta, G., Deerinck, T.J., Adams, S.R., Bouwer, J., Tour, O., Laird, D.W., Sosinsky, G.E., Tsien, R.Y., Ellisman, M.H., Multicolor and electron microscopic imaging of connexin trafficking. Science 296 (2002), 503–507 https://doi.org/10.1126/science.1068793.
George, C.H., Kendall, J.M., Evans, W.H., Intracellular trafficking pathways in the assembly of connexins into gap junctions. J. Biol. Chem. 274 (1999), 8678–8685 https://doi.org/10.1074/jbc.274.13.8678.
Grieve, A.G., Rabouille, C., Golgi bypass: skirting around the heart of classical secretion. Cold Spring Harb. Perspect. Biol., 3, 2011, a005298 https://doi.org/10.1101/cshperspect.a005298.
Grifa, A., Wagner, C.A., D'Ambrosio, L., Melchionda, S., Bernardi, F., Lopez-Bigas, N., Rabionet, R., Arbones, M., Monica, M.D., Estivill, X., et al. Mutations in GJB6 cause nonsyndromic autosomal dominant deafness at DFNA3 locus. Nat. Genet. 23 (1999), 16–18 https://doi.org/10.1038/12612.
Hervé, J.C., Derangeon, M., Sarrouilhe, D., Bourmeyster, N., Influence of the scaffolding protein Zonula Occludens (ZOs) on membrane channels. Biochim. Biophys. Acta 1838 (2014), 595–604 https://doi.org/10.1016/j.bbamem.2013.07.006.
Hoang Dinh, E., Ahmad, S., Chang, Q., Tang, W., Stong, B., Lin, X., Diverse deafness mechanisms of connexin mutations revealed by studies using in vitro approaches and mouse models. Brain Res. 1277 (2009), 52–69 https://doi.org/10.1016/j.brainres.2009.02.008.
Hung, A., Yarovsky, I., Gap junction hemichannel interactions with zwitterionic lipid, anionic lipid, and cholesterol: molecular simulation studies. Biochemistry 50 (2011), 1492–1504 https://doi.org/10.1021/bi1004156.
Hunter, A.W., Barker, R.J., Zhu, C., Gourdie, R.G., Zonula occludens-1 alters connexin43 gap junction size and organization by influencing channel accretion. Mol. Biol. Cell 16 (2005), 5686–5698 https://doi.org/10.1091/mbc.E05-08-0737.
Jacob, R., Heine, M., Alfalah, M., Naim, H.Y., Distinct cytoskeletal tracks direct individual vesicle populations to the apical membrane of epithelial cells. Curr. Biol. 13 (2003), 607–612 https://doi.org/10.1016/S0960-9822(03)00188-X.
Jagger, D.J., Forge, A., Connexins and gap junctions in the inner ear-it's not just about K+ recycling. Cell Tissue Res. 360 (2015), 633–644 https://doi.org/10.1007/s00441-014-2029-z.
Janes, P.W., Ley, S.C., Magee, A.I., Aggregation of lipid rafts accompagnies signaling via the T cell antigen receptor. J. Cell Biol. 147 (1999), 447–461 https://doi.org/10.1083/jcb.147.2.447.
Kamiya, K., Yum, S.W., Kurebayashi, N., Muraki, M., Ogawa, K., Karasawa, K., Miwa, A., Guo, X., Gotoh, S., Sugitani, Y., et al. Assembly of the cochlear gap junction macromolecular complex requires connexin 26. J. Clin. Invest. 124 (2014), 1598–1607 https://doi.org/10.1172/JCI67621.
Kelsell, D.P., Dunlop, J., Stevens, H.P., Lench, N.J., Liang, J.N., Parry, G., Mueller, R.F., Leigh, I.M., Connexin 26 mutations in hereditary non-syndromic sensorineural deafness. Nature 387 (1997), 80–83 https://doi.org/10.1038/387080a0.
Kikuchi, T., Kimura, R.S., Paul, D.L., Takasaka, T., Adams, J.C., Gap junction systems in the mammalian cochlea. Brain Res. Brain Res. Rev. 32 (2000), 163–166 https://doi.org/10.1016/S0165-0173(99)00076-4.
Kotini, M., Mayor, R., Connexins in migration during development and cancer. Dev. Biol. 401 (2015), 143–151 https://doi.org/10.1016/j.ydbio.2014.12.023.
Korver, A.M., Smith, R.J., Van Camp, G., Schleiss, M.R., Bitner-Glindzicz, M.A., Lustig, L.R., Usami, S.I., Boudewyns, A.N., Congenital hearing loss. Nat. Rev. Dis. Primers, 3, 2017, 16094 https://doi.org/10.1038/nrdp.2016.94.
Koval, M., Harley, J.E., Hick, E., Steinberg, H.E., Connexin46 is retained as monomers in a trans-Golgi compartment of osteoblastic cells. J. Cell Biol. 137 (1997), 847–857 https://doi.org/10.1083/jcb.137.4.847.
Laird, D.W., Life cycle of connexins in health and disease. Biochem. J. 394 (2006), 527–543 https://doi.org/10.1042/BJ20051922.
Lauf, U., Giepmans, B.N., Lopez, P., Braconnot, S., Chen, S.C., Falk, M.M., Dynamic trafficking and delivery of connexons to the plasma membrane and accretion to gap junctions in living cells. Proc. Natl. Acad. Sci. U.S.A. 99 (2002), 10446–10451 https://doi.org/10.1073/pnas.162055899.
Lilly, E., Sellitto, C., Milstone, L.M., White, T.W., Connexin channels in congenital skin disorders. Semin. Cell Dev. Biol. 50 (2016), 4–12 https://doi.org/10.1016/j.semcdb.2015.11.018.
Locke, D., Harris, A.L., Connexin channels and phospholipids: association and modulation. BMC Biol., 7, 2009, 52 https://doi.org/10.1186/1741-7007-7-52.
Locke, D., Liu, J., Harris, A.L., Lipid rafts prepared by different methods contain different connexin channels, but gap junction are not lipid rafts. Biochemistry 44 (2005), 13027–13042 https://doi.org/10.1021/bi050495a.
Martin, P.E., Blundell, G., Ahmad, S., Errington, R.J., Evans, W.H., Multiple pathways in the trafficking and assembly of connexin 26, 32 and 43 into gap junction intercellular communication channels. J. Cell Sci. 114 (2001), 3845–3855.
Musil, L.S., Goodenough, D.A., Biochemical analysis of connexin43 intracellular transport, phosphorylation, and assembly into gap junctional plaques. J. Cell Biol. 115 (1991), 1357–1374 https://doi.org/10.1083/jcb.115.5.1357.
Musil, L.S., Goodenough, D.A., Multisubunit assembly of an integral plasma membrane channel protein, gap junction connexin43, occurs after exit from the ER. Cell 74 (1993), 1065–1077 https://doi.org/10.1016/0092-8674(93)90728-9.
Pike, L.J., Lipid rafts: bringing order to chaos. J. Lipid Res. 44 (2003), 655–667 https://doi.org/10.1194/jlr.R200021-JLR200.
Schliwa, M., Action of cytochalasin D on cytoskeletal networks. J. Cell Biol. 92 (1982), 79–91 https://doi.org/10.1083/jcb.92.1.79.
Schubert, A.L., Schubert, W., Spray, D.C., Lisanti, M.P., Connexin family members target to lipid raft domains and interact with caveolin-1. Biochemistry 41 (2002), 5754–5764 https://doi.org/10.1021/bi0121656.
Sheng, M., Sala, C., PDZ domains and the organization of supramolecular complexes. Annu. Rev. Neurosci. 24 (2001), 1–29 https://doi.org/10.1146/annurev.neuro.24.1.1.
Sun, J., Ahmad, S., Chen, S., Tang, W., Zhang, Y., Chen, P., Lin, X., Cochlear gap junctions coassembled from Cx26 and 30 show faster intercellular Ca2+ signaling than homomeric counterparts. Am. J. Physiol. Cell Physiol. 288 (2005), C613–C623 https://doi.org/10.1152/ajpcell.00341.2004.
Thévenin, A.F., Kowal, T.J., Fong, J.T., Kells, R.M., Fisher, C.G., Falk, M.M., Proteins and mechanisms regulating gap-junction assembly, internalization, and degradation. Physiology (Bethesda) 28 (2013), 93–116 https://doi.org/10.1152/physiol.00038.2012.
Traub, O., Look, J., Paul, D., Willecke, K., Cyclic adenosine monophosphate stimulates biosynthesis and phosphorylation of the 26 kDa gap junction protein in cultured mouse hepatocytes. Eur. J. Cell Biol. 43 (1987), 48–54.
Walsh, T., Pierce, S.B., Lenz, D.R., Brownstein, Z., Dagan-Rosenfeld, O., Shahin, H., Roeb, W., McCarthy, S., Nord, A.S., Gordon, C.R., et al. Genomic duplication and overexpression of TJP2/ZO-2 leads to altered expression of apoptosis genes in progressive nonsyndromic hearing loss DFNA51. Am. J. Hum. Genet. 87 (2010), 101–109 https://doi.org/10.1016/j.ajhg.2010.05.011.
Xu, J., Nicholson, B.J., The role of connexins in ear and skin physiology – functional insights from disease-associated mutations. Biochim. Biophys. Acta 1828 (2013), 167–178 https://doi.org/10.1016/j.bbamem.2012.06.024.
Zhang, J.T., Chen, M., Foote, C.I., Nicholson, B.J., Membrane integration of in vitro-translated gap junctional proteins: co- and post-translational mechanisms. Mol. Biol. Cell 7 (1996), 471–482.