Mammalian prestin is a weak Cl⁻/HCO₃⁻ electrogenic antiporter.
The lateral membrane of mammalian cochlear outer hair cells contains prestin, a protein which can act as a fast voltage-driven actuator responsible for electromotility and enhanced sensitivity to sound. The protein belongs to the SLC26 family of transporters whose members are characterised as able to exchange halides for SO(4)(2-) or HCO(3)(-) yet previous analyses of mammalian prestin have suggested that such exchange functions were minimal. Here anion transport is investigated both in guinea-pig outer hair cells (OHCs) and in an expression system where we employ a sensitive intracellular pH (pH(i)) probe, pHluorin, to report HCO(3)(-) transport and to monitor the small pH(i) changes observable in the cells. In the presence of extracellular HCO(3)(-), pH(i) recovered from an acid load 4 times faster in prestin-transfected cells. The acceleration required a chloride gradient established by reducing extracellular chloride to 2 mm. Similar results were also shown using BCECF as an alternative pH(i) sensor, but with recovery only found in those cells expressing prestin. Simultaneous electrophysiological recording of the transfected cells during bicarbonate exposure produced a shift in the reversal potential to more negative potentials, consistent with electrogenic transport. These data therefore suggest that prestin can act as a weak Cl(-)/HCO(3)(-) antiporter and it is proposed that, in addition to participating in wide band cochlear sound amplification, prestin may also be involved in the slow time scale (>10 s) phenomena where changes in cell stiffness and internal pressure have been implicated. The results show the importance of considering the effects of the endogenous bicarbonate buffering system in evaluating the function of prestin in cochlear outer hair cells.
UCL Ear Institute, 332 Gray’s Inn Road, London WC1E 6BT, UK.
P
P
Mistrík
N
N
Daudet
K
K
Morandell
J F
JF
Ashmore
D000200:Action Potentials; D000818:Animals; D027321:Anion Transport Proteins / Q000235:genetics / Q000502:physiology*; D001639:Bicarbonates / Q000378:metabolism; D016466:CHO Cells; D027963:Chloride-Bicarbonate Antiporters / Q000502:physiology*; D002713:Chlorine / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006168:Guinea Pigs; D018072:Hair Cells, Auditory, Outer / Q000502:physiology; D006863:Hydrogen-Ion Concentration; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D000076988:Sulfate Transporters
D027321:Anion Transport Proteins; D001639:Bicarbonates; D027963:Chloride-Bicarbonate Antiporters; C485187:PHluorin; D011993:Recombinant Fusion Proteins; C409207:Slc26a5 protein, rat; D000076988:Sulfate Transporters; D049452:Green Fluorescent Proteins; D002713:Chlorine