Abstract
The Tmhs gene codes for a tetraspan transmembrane protein that is expressed in hair cell stereocilia. We previously showed that a spontaneous missense mutation of Tmhs underlies deafness and vestibular dysfunction in the hurry-scurry (hscy) mouse. Subsequently, mutations in the human TMHS gene were shown to be responsible for DFNB67, an autosomal recessive nonsyndromic deafness locus. Here we describe a genetically engineered null mutation of the mouse Tmhs gene (Tmhs tm1Kjn) and show that its phenotype is identical to that of the hscy missense mutation, confirming the deleterious nature of the hscy cysteine-to-phenylalanine substitution. In the targeted null allele, the Tmhs promoter drives expression of a lacZ reporter gene. Visualization of β-galactosidase activity in Tmhs tm1Kjn heterozygous mice indicates that Tmhs is highly expressed in the cochlear and vestibular hair cells of the inner ear. Expression is first detectable at E15.5, peaks around P0, decreases slightly at P6, and is absent by P15, a duration that supports the involvement of Tmhs in stereocilia development. Tmhs reporter gene expression also was detected in several cranial and cervical sensory ganglia, but not in the vestibular or spiral ganglia. We also describe a new nontargeted mutation of the Tmhs gene, hscy-2J, that causes abnormal splicing from a cryptic splice site within exon 2 and is predicted to produce a functionally null protein lacking 51 amino acids of the wild-type sequence.
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Acknowledgments
The authors thank the following personnel of the Jackson Laboratory for their contributions to this article: Patsy Nishina and David Bergstrom for helpful comments on the manuscript, Sandra Gray for mouse colony management, Heping Yu for ABR technical assistance, Peter Finger for SEM assistance, Heather Carlisle for design and assembly of the gene-targeting construct, and personnel of the Cell Biology and Microinjection Services for generating the B6.129-Tmhs tm1Kjnmice. They also thank the personnel of the Neuroscience Mutagenesis Facility and the Fine Mapping Service for the initial phenotyping and mapping of the nmf430 (Tmhs hscy-2J) mutation. This work was supported by National Institutes of Health (NIH) Grants DC004301, RR01183, and DC005590 (BF). The Jackson Laboratory institutional shared services are supported by NIH Grant CA34196.
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Longo-Guess, C.M., Gagnon, L.H., Fritzsch, B. et al. Targeted knockout and lacZ reporter expression of the mouse Tmhs deafness gene and characterization of the hscy-2J mutation. Mamm Genome 18, 646–656 (2007). https://doi.org/10.1007/s00335-007-9049-x
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DOI: https://doi.org/10.1007/s00335-007-9049-x