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Three ENU-induced neurological mutations in the pore loopof sodium channel Scn8a (Nav1.6) and a genetically linkedretinal mutation, rd13

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Abstract

The goal of The Jackson Laboratory Neuroscience Mutagenesis Facility is to generate mouse models of human neurological disease. We describe three new models obtained from a three-generation screen for recessive mutations. Homozygous mutant mice from lines nmf2 and nmf5 exhibit hind limb paralysis and juvenile lethality. Homozygous nmf58 mice exhibit a less severe movement disorder that includes sustained dystonic postures. The mutations were mapped to the distal region of mouse Chromosome (Chr) 15. Failure to complement a mutant allele of a positional candidate gene, Scn8a, demonstrated that the mutations are new alleles of Scn8a. Missense mutations of evolutionarily conserved residues of the sodium channel were identified in the three lines, with the predicted amino acid substitutions N1370T, I1392F, and L1404H. These residues are located within the pore loop of domain 3 of sodium channel Nav1.6. The lethal phenotypes suggest that the new alleles encode proteins with partial or complete loss of function. Several human disorders are caused by mutation in the pore loop of domain 3 of paralogous sodium channel genes. Line nmf5 contains a second, independent mutation in the rd13 locus that causes a reduction in cell number in the outer nuclear layer of the retina. rd13 was mapped to the distal 4 Mb of Chr 15. No coding or splice site mutations were detected in Pde1b, a candidate gene for rd13. The generation of three independent Scn8a mutations among 1100 tested G3 families demonstrates that the Scn8a locus is highly susceptible to ENU mutagenesis. The new alleles of Scn8a will be valuable for analysis of sodium channel physiology and disease.

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Acknowledgments

We acknowledge the contributions of Louise Dionne for recognition of the mutant phenotypes in lines nmf2, nmf5, and nmf58; Gregory Cox for advice on complementation testing; Joseph Brzezinski for assistance with retinal histology; and Maja Adamska for genomic sequencing of Pde1b. This work was supported by the Neuroscience Mutagenesis Facility at The Jackson Laboratory (U01 NS41215), the Cancer Center Support Grant to The Jackson Laboratory (CA34196), and research grant R01 NS34509 (M.H. Meisler). D.A. Buchner acknowledges support from the Hearing and Chemical Senses Training Program (T32 DC00011). Genotyping was provided by The Jackson Laboratory Allele Typing Service.

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  1. Department of Human Genetics, University of Michigan, 4708 Medical Science II, Ann Arbor, Michigan, 48109-0618, USA

    David A. Buchner & Miriam H. Meisler

  2. The Jackson Laboratory, Bar Harbor, Maine, 04609, USA

    Kevin L. Seburn & Wayne N. Frankel

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  1. David A. Buchner
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  2. Kevin L. Seburn
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Correspondence to Miriam H. Meisler.

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(David A. Buchner and Kevin L. Seburn) These authors contributed equally.

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Buchner, D.A., Seburn, K.L., Frankel, W.N. et al. Three ENU-induced neurological mutations in the pore loopof sodium channel Scn8a (Nav1.6) and a genetically linkedretinal mutation, rd13 . Mamm Genome 15, 344–351 (2004). https://doi.org/10.1007/s00335-004-2332-1

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