Myosin VIIA is required for aminoglycoside accumulation in cochlear hair cells.
Myosin VIIA is expressed by sensory hair cells and has a primary structure predicting a role in membrane trafficking and turnover, processes that may underlie the susceptibility of hair cells to aminoglycoside antibiotics. [3H]Gentamicin accumulation and the effects of aminoglycosides were therefore examined in cochlear cultures of mice with different missense mutations in the myosin VIIA gene, Myo7a, to see whether myosin VIIA plays a role in aminoglycoside ototoxicity. Hair cells from homozygous mutant Myo7ash1 mice, with a mutation in a nonconserved region of the myosin VIIA head, respond rapidly to aminoglycoside treatment and accumulate high levels of gentamicin. Hair cells from homozygous mutant Myo7a6J mice, with a mutation at a highly conserved residue close to the ATP binding site of the myosin VIIA head, do not accumulate [3H]gentamicin and are protected from aminoglycoside ototoxicity. Hair cells from heterozygotes of both alleles accumulate [3H]gentamicin and respond to aminoglycosides. Although aminoglycoside uptake is thought to be via apical surface-associated endocytosis, coated pit numbers on the apical membrane of heterozygous and homozygous Myo7a6J hair cells are similar. Pulse-chase experiments with cationic ferritin confirm that the apical endocytotic pathway is functional in homozygous Myo7a6J hair cells. Transduction currents can be recorded from both heterozygous and homozygous Myo7a6J hair cells, suggesting it is unlikely that the drug enters via diffusion through the mechanotransducer channel. The results show that myosin VIIA is required for aminoglycoside accumulation in hair cells. Myosin VIIA may transport a putative aminoglycoside receptor to the hair cell surface, indirectly translocate it to sites of membrane retrieval, or retain it in the endocytotic pathway.
Richardson
G P
GP
School of Biological Sciences, University of Sussex, Brighton BN1 9QG, United Kingdom.
Forge
A
A
Kros
C J
CJ
Fleming
J
J
Brown
S D
SD
Steel
K P
KP
D000255:Adenosine Triphosphate / Q000378:metabolism; D019943:Amino Acid Substitution / Q000502:physiology; D000818:Animals; D000831:Animals, Newborn; D000900:Anti-Bacterial Agents / Q000009:adverse effects / Q000493:pharmacokinetics*; D001665:Binding Sites / Q000502:physiology; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D002478:Cells, Cultured; D004398:Dyneins; D004705:Endocytosis / Q000187:drug effects; D005614:Freeze Fracturing; D005839:Gentamicins / Q000009:adverse effects / Q000493:pharmacokinetics*; D018072:Hair Cells, Auditory, Outer / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D000081412:Myosin VIIa; D009218:Myosins / Q000235:genetics* / Q000378:metabolism; D009355:Neomycin / Q000009:adverse effects / Q000493:pharmacokinetics*; D017354:Point Mutation; D014316:Tritium
D000900:Anti-Bacterial Agents; D005839:Gentamicins; C000631456:Myo7a protein, mouse; D000081412:Myosin VIIa; D014316:Tritium; D000255:Adenosine Triphosphate; D009218:Myosins; D004398:Dyneins; D009355:Neomycin