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Targeted knockout and lacZ reporter expression of the mouse Tmhs deafness gene and characterization of the hscy-2J mutation
Journal article   Peer reviewed

Targeted knockout and lacZ reporter expression of the mouse Tmhs deafness gene and characterization of the hscy-2J mutation

Chantal M Longo-Guess, Leona H Gagnon, Bernd Fritzsch and Kenneth R Johnson
Mammalian genome, Vol.18(9), pp.646-656
09/2007
DOI: 10.1007/s00335-007-9049-x
PMCID: PMC2613174
PMID: 17876667
url
https://www.ncbi.nlm.nih.gov/pmc/articles/2613174View
Open Access

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 beta-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.
 Amino Acid Sequence Gene Targeting Sequence Deletion Deafness - genetics RNA Splice Sites - genetics Membrane Proteins - genetics Alternative Splicing - genetics Humans Mice, Inbred C57BL Deafness - physiopathology Exons - genetics Male Mice, Knockout Membrane Proteins - deficiency Animals Membrane Proteins - chemistry Mutagenesis Mice, Mutant Strains Female Mice Mice, Inbred BALB C Genes, Reporter Lac Operon

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