Andersen’s syndrome mutants produce a knockdown of inwardly rectifying K+ channel in mouse skeletal muscle in vivo
- 204 Downloads
Andersen’s syndrome (AS) is a rare autosomal disorder that has been defined by the triad of periodic paralysis, cardiac arrhythmia, and developmental anomalies. AS has been directly linked to over 40 different autosomal dominant negative loss-of-function mutations in the KCNJ2 gene, encoding for the tetrameric strong inward rectifying K+ channel KIR2.1. While KIR2.1 channels have been suggested to contribute to setting the resting membrane potential (RMP) and to control the duration of the action potential (AP) in skeletal and cardiac muscle, the mechanism by which AS mutations produce such complex pathophysiological symptoms is poorly understood. Thus, we use an adenoviral transduction strategy to study in vivo subcellular distribution of wild-type (WT) and AS-associated mutant KIR2.1 channels in mouse skeletal muscle. We determined that WT and D71V AS mutant KIR2.1 channels are localized to the sarcolemma and the transverse tubules (T-tubules) of skeletal muscle fibers, while the ∆314-315 AS KIR2.1 mutation prevents proper trafficking of the homo- or hetero-meric channel complexes. Whole-cell voltage-clamp recordings in individual skeletal muscle fibers confirmed the reduction of inwardly rectifying K+ current (IK1) after transduction with ∆314-315 KIR2.1 as compared to WT channels. Analysis of skeletal muscle function revealed reduced force generation during isometric contraction as well as reduced resistance to muscle fatigue in extensor digitorum longus muscles transduced with AS mutant KIR2.1. Together, these results suggest that KIR2.1 channels may be involved in the excitation–contraction coupling process required for proper skeletal muscle function. Our findings provide clues to mechanisms associated with periodic paralysis in AS.
KeywordsAndersen’s syndrome KIR2.1 Skeletal muscle Adenovirus Channelopathies
This work was supported by the Centre National de la Recherche Scientifique (CNRS) and Association Française contre les Myopathies (AFM) grant (S.B.) and Chateaubriand fellowships (D.S.) and City of Nice fellowship (S.G. and A.V.). We thank the Vector Core of the University Hospital of Nantes for providing the adenovirus vectors.
Conflict of interest
The authors declare having no conflict of interest.
- Allen DG, Lamb GD, Westerblad H (2008a) Impaired calcium release during fatigue. J Appl Physiol 104:296-305Google Scholar
- Allen DG, Lamb GD, Westerblad H (2008b) Skeletal muscle fatigue: cellular mechanisms. Physiol Rev 88:287-332Google Scholar
- Allen DG, Westerblad H, Lee JA, Lannergren J (1992) Role of excitation-contraction coupling in muscle fatigue. Sports Med 13:116-126Google Scholar
- Ashen MD, O'Rourke B, Kluge KA, Johns DC, Tomaselli GF (1995) Inward rectifier K+ channel from human heart and brain: cloning and stable expression in a human cell line. Am J Phys 268:H506–H511Google Scholar
- Ballester LY, Vanoye CG, George AL, Jr. (2007) Exaggerated Mg2+ inhibition of Kir2.1 As a consequence of reduced PIP2 sensitivity in Andersen syndrome. Channels (Austin) 1:209-217Google Scholar
- Donaldson MR, Jensen JL, Tristani-Firouzi M, Tawil R, Bendahhou S, Suarez WA, Cobo AM, Poza JJ, Behr E, Wagstaff J, Szepetowski P, Pereira S, Mozaffar T, Escolar DM, Fu YH, Ptacek LJ (2003) PIP2 Binding residues of Kir2.1 Are common targets of mutations causing Andersen syndrome. Neurology 60:1811–1816CrossRefPubMedGoogle Scholar
- Inagaki N, Tsuura Y, Namba N, Masuda K, Gonoi T, Horie M, Seino Y, Mizuta M, Seino S (1995) Cloning and functional characterization of a novel ATP-sensitive potassium channel ubiquitously expressed in rat tissues, including pancreatic islets, pituitary, skeletal muscle, and heart. J Biol Chem 270:5691–5694CrossRefPubMedGoogle Scholar
- Kokunai Y, Nakata T, Furuta M, Sakata S, Kimura H, Aiba T, Yoshinaga M, Osaki Y, Nakamori M, Itoh H, Sato T, Kubota T, Kadota K, Shindo K, Mochizuki H, Shimizu W, Horie M, Okamura Y, Ohno K, Takahashi MP (2014) A Kir3.4 Mutation causes Andersen-Tawil syndrome by an inhibitory effect on Kir2.1. Neurology 82:1058–1064CrossRefPubMedGoogle Scholar
- Kristensen M, Hansen T, Juel C (2006) Membrane proteins involved in potassium shifts during muscle activity and fatigue. Am J Physiol 290:R766–R772Google Scholar
- Plaster NM, Tawil R, Tristani-Firouzi M, Canun S, Bendahhou S, Tsunoda A, Donaldson MR, Iannaccone ST, Brunt E, Barohn R, Clark J, Deymeer F, George AL Jr, Fish FA, Hahn A, Nitu A, Ozdemir C, Serdaroglu P, Subramony SH, Wolfe G, Fu YH, Ptacek LJ (2001) Mutations in Kir2.1 Cause the developmental and episodic electrical phenotypes of Andersen's syndrome. Cell 105:511–519CrossRefPubMedGoogle Scholar
- Priori SG, Pandit SV, Rivolta I, Berenfeld O, Ronchetti E, Dhamoon A, Napolitano C, Anumonwo J, di Barletta MR, Gudapakkam S, Bosi G, Stramba-Badiale M, Jalife J (2005) A novel form of short QT syndrome (SQT3) is caused by a mutation in the KCNJ2 gene. Circ Res 96:800–807CrossRefPubMedGoogle Scholar
- Reiken S, Lacampagne A, Zhou H, Kherani A, Lehnart SE, Ward C, Huang F, Gaburjakova M, Gaburjakova J, Rosemblit N, Warren MS, He KL, Yi GH, Wang J, Burkhoff D, Vassort G, Marks AR (2003) PKA phosphorylation activates the calcium release channel (ryanodine receptor) in skeletal muscle: defective regulation in heart failure. J Cell Biol 160:919–928CrossRefPubMedPubMedCentralGoogle Scholar
- Ryan DP, da Silva MR, Soong TW, Fontaine B, Donaldson MR, Kung AW, Jongjaroenprasert W, Liang MC, Khoo DH, Cheah JS, Ho SC, Bernstein HS, Maciel RM, Brown RH Jr, Ptacek LJ (2010) Mutations in potassium channel Kir2.6 Cause susceptibility to thyrotoxic hypokalemic periodic paralysis. Cell 140:88–98CrossRefPubMedPubMedCentralGoogle Scholar
- Sacco S, Giuliano S, Sacconi S, Desnuelle C, Barhanin J, Amri EZ, Bendahhou S (2014) The inward rectifier potassium channel Kir2.1 is required for osteoblastogenesis. Hum Mol Genet 24:471–479Google Scholar
- Sakura H, Ammala C, Smith PA, Gribble FM, Ashcroft FM (1995a) Cloning and functional expression of the cDNA encoding a novel ATP-sensitive potassium channel subunit expressed in pancreatic beta-cells, brain, heart and skeletal muscle. FEBS Lett 377:338-344Google Scholar
- Sakura H, Bond C, Warren-Perry M, Horsley S, Kearney L, Tucker S, Adelman J, Turner R, Ashcroft FM (1995b) Characterization and variation of a human inwardly-rectifying-K-channel gene (KCNJ6): a putative ATP-sensitive K-channel subunit. FEBS Lett 367:193-197Google Scholar
- Tristani-Firouzi M, Jensen JL, Donaldson MR, Sansone V, Meola G, Hahn A, Bendahhou S, Kwiecinski H, Fidzianska A, Plaster N, Fu YH, Ptacek LJ, Tawil R (2002) Functional and clinical characterization of KCNJ2 mutations associated with LQT7 (Andersen syndrome). J Clin Invest 110:381–388CrossRefPubMedPubMedCentralGoogle Scholar