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K H Iwasa

Publications and source records attributed to K H Iwasa.

At least 19 recordsLinked to original sources
PubMed2006

Effects of tarantula toxin GsMTx4 on the membrane motor of outer hair cells.

GsMTx4, a cationic hydrophobic peptide isolated from tarantula venom, is a specific inhibitor of stretch-activated channels (SACs). Here, we show that the toxin also affects the membrane motor of outer hair cells at low doses. The membrane motor of outer hair cells is based on prestin, a member of the SLC26 family of membrane proteins, and directly uses electrical energy available at the plasma membrane. It is considered to be an essential part of the "cochlear amplifier," which increases the sensitivity, tuning, and dynamic range of the mammalian ear. The toxin shifts the operating point of the motor. The saturating value of the voltage shift is (26 +/- 1) mV, capable of significantly reducing the performance of the cochlear amplifier. The dissociation constant is (3.1 +/- 0.6) microM, about five-fold higher than that for SACs.

Animals↗
PubMed2004

Tension sensitivity of prestin: comparison with the membrane motor in outer hair cells.

The membrane motor in outer hair cells undergoes conformational transitions involving charge displacement of approximately 0.8 e across the membrane and changes of approximately 4 nm(2) in its membrane area. Previous reports have established that the charge transfer in the membrane motor and that in prestin, a membrane protein in the plasma membrane of outer hair cells, are approximately equal. Here, we determine the membrane area changes based on its sensitivity to membrane tension. We found that prestin does undergo area changes and that the magnitude is approximately 1 nm(2), smaller than the value 4 nm(2) for outer hair cell motor. This result confirms that prestin is a protein that functions as a membrane motor based on piezoelectricity. The discrepancy in the magnitude could suggest a prestin-containing complex in outer hair cells.

Anion Transport Proteins↗
PubMed2002

Cooperative interaction as the physical basis of the negative stiffness in hair cell stereocilia.

A recent report confirmed that stiffness of the stereocilia can be negative, as predicted by the Howard-Hudspeth model. According to this model, the mechanotransducer channel's gating not only reduces the stereociliary stiffness, but can alter its sign as well. The basic assumptions of this model do not include cooperativity in channel gating. Here we consider two possible explanations for the observed negative stiffness. If the stereocilia have a special structure so that microscopic displacement can be imposed on each channel by controlling the bending of the bundle, negative stiffness can occur without channel cooperativity. If such a microscopic condition cannot be imposed by a macroscopic manipulation, an additional physical process, such as cooperativity in channel gating, is required to explain negative stiffness.

Hair Cells, Auditory↗
PubMed2001

A two-state piezoelectric model for outer hair cell motility.

Recent studies have revealed that voltage-dependent length changes of the outer hair cell are based on charge transfer across the membrane. Such a motility can be explained by an "area motor" model, which assumes two states in the motor and that conformational transitions involve transfer of motor charge across the membrane and mechanical displacements of the membrane. Here it is shown that the area motor is piezoelectric and that the hair cell that incorporates such a motor in its lateral membrane is also piezoelectric. Distinctive features of the outer hair cell are its exceptionally large piezoelectric coefficient, which exceeds the best known piezoelectric material by four orders of magnitude, and its prominent nonlinearity due to the discreteness of motor states.

Animals↗
PubMed2000

Fluctuation of motor charge in the lateral membrane of the cochlear outer hair cell.

Functioning of the membrane motor of the outer hair cell is tightly associated with transfer of charge across the membrane. To obtain further insights into the motor mechanism, we examined kinetics of charge transfer across the membrane in two different modes. One is to monitor charge transfer induced by changes in the membrane potential as an excess membrane capacitance. The other is to measure spontaneous flip-flops of charges across the membrane under voltage-clamp conditions as current noise. The noise spectrum of current was inverse Lorentzian, and the capacitance was Lorentzian, as theoretically expected. The characteristic frequency of the capacitance was approximately 10 kHz, and that for current noise was approximately 30 kHz. The difference in the characteristic frequencies seems to reflect the difference in the modes of mechanical movement associated with the two physical quantities.

Animals↗
PubMed2000

Effect of turgor pressure on outer hair cell motility.

The motor in the outer hair cell converts electrical energy directly into mechanical energy. There are two possible mechanisms for such a motor: one depends on changes in the membrane area ("area motor") and the other on changes in the stiffness ("stiffness motor"). These two mechanisms, which are not mutually exclusive, give different predictions on turgor pressure dependence of the amplitude. It was found that an increased pressure shifts the voltage dependence but does not change the amplitude of length changes. This observation is incompatible with a pure stiffness motor model. It was also confirmed that length changes are closely related to charge movements by monitoring the membrane capacitance. It can be concluded that hair cell motility is primarily based on area changes, and not changes in the elastic moduli.

Animals↗
PubMed1999

Electrically driven motor in the outer hair cell: effect of a mechanical constraint.

The outer hair cell has a unique voltage-dependent motility associated with charge transfer across the plasma membrane. To examine mechanical changes in the membrane that are coupled with such charge movements, we digested the undercoating of the membrane with trypsin. We inflated the cell into a sphere and constrained the surface area by not allowing volume changes. We found that this constraint on the membrane area sharply reduced motor-associated charge movement across the membrane, demonstrating that charge transfer is directly coupled with membrane area change. This electromechanical coupling in the plasma membrane must be the key element for the motile mechanism of the outer hair cell.

Animals↗
PubMed1998

Prenatal expression of inwardly rectifying potassium channel mRNA (Kir4.1) in rat brain.

The levels and cellular localization of the mRNA encoding the inwardly rectifying potassium ion channel Kir4.1 were investigated in the embryonic rat brain by Northern blots and in situ hybridization. This transcript was absent at embryonic day 13 (E13), whereas it was clearly present in E14-15 preparations, principally in the neuroepithelium of the cerebral cortex, thalamus, and hypothalamus. At later embryonic stages (E17-20), Kir4.1 mRNA levels increased and expanded to the mantle zone, such as the cortical plate, hippocampus, thalamus, and hypothalamus. The early appearance of Kir4.1 mRNA in various brain regions suggests an involvement of the channel in cell proliferation, migration and differentiation in the rat CNS.

Animals↗
PubMed1997

Force generation in the outer hair cell of the cochlea.

The outer hair cell of the mammalian cochlea has a unique motility directly dependent on the membrane potential. Examination of the force generated by the cell is an important step in clarifying the detailed mechanism as well as the biological importance of this motility. We performed a series of experiments to measure force in which an elastic probe was attached to the cell near the cuticular plate and the cell was driven with voltage pulses delivered from a patch pipette under whole-cell voltage clamp. The axial stiffness was also determined with the same cell by stretching it with the patch pipette. The isometric force generated by the cell is around 0.1 nN/mV, somewhat smaller than 0.15 nN/mV, predicted by an area motor model based on mechanical isotropy, but larger than in earlier reports in which the membrane potential was not controlled. The axial stiffness obtained, however, was, on average, 510 nN per unit strain, about half of the value expected from the mechanical isotropy of the membrane. We extended the area motor theory incorporating mechanical orthotropy to accommodate the axial stiffness determined. The force expected from the orthotropic model was within experimental uncertainties.

Animals↗
PubMed1997

Effect of diamide on force generation and axial stiffness of the cochlear outer hair cell.

We found that diamide, which affects spectrin, reduces the axial stiffness of the cochlear outer hair cell, the cylindrically shaped mechanoreceptor cell with a unique voltage-sensitive motility. This effect thus provides a means of examining the relationship between the stiffness and the motility of the cell. For measuring axial stiffness and force production, we used an experimental configuration in which an elastic probe was attached to the cell near the cuticular plate and the other end of the cell was held with a patch pipette in the whole-cell recording mode. Diamide at concentrations of up to 5 mM reduced the axial stiffness in a dose-dependent manner to 165 nN per unit strain from 502 nN for untreated cells. The isometric force elicited by voltage pulses under whole-cell voltage clamp was also reduced to 35 pN/mV from 105 pN/mV for untreated cells. Thus the isometric force was approximately proportional to the axial stiffness. Our observations suggest a series connection between the motor and cytoskeletal elements and can be explained by the area motor model previously proposed for the outer hair cell.

Animals↗
PubMed1997

Current noise spectrum and capacitance due to the membrane motor of the outer hair cell: theory.

The voltage-dependent motility of the outer hair cell is based on a membrane motor densely distributed in the lateral membrane. The gating charge of the membrane motor is manifested as a bell-shaped membrane potential dependence of the membrane capacitance. In this paper it is shown that movements of the gating charge should produce a high-pass current noise described by an inverse Lorentzian similar to the one shown by Kolb and Läuger for ion carriers. The frequency dependence of the voltage-dependent capacitance is also derived. These derivations are based on membrane motor models with two or three states. These two models lead to similar predictions on the capacitance and current noise. It is expected that the examination of the spectral properties of these quantities would be a useful means of determining the relaxation time for conformational transitions of the membrane motor.

Animals↗
PubMed1997

Ultrastructural localization of the Na-K-Cl cotransporter in the lateral wall of the rabbit cochlear duct.

Localization of the immunoreactivity in the lateral wall of the rabbit cochlear duct was examined using a post-embedding immunogold method with a polyclonal antiserum raised against the rabbit parotid Na-K-Cl cotransporter. In the stria vascularis, the labeling was significant on the basolateral membrane infolding of marginal cells, whereas no labeling was seen on the luminal membrane of these cells. Immunoreactivity was also detected on the cell membranes of various other cells. These include fibrocytes of the spiral ligament and the spiral prominence, and vascular endothelial cells in the stria vascularis and the spiral ligament. In contrast, virtually no gold particles were seen on the membrane of intermediate cells, basal cells of the stria vascularis, the epithelial cells of the spiral prominence, or Reissner's membrane. Our result on the localization of the Na-K-Cl cotransporter in marginal cells is consistent with electrophysiological studies (Wangemann et al. (1995) Hear. Res. 84, 19-29). Our result on fibrocytes is discussed in relation to K+ circulation into endolymph from perilymph (Schulte and Steel (1994) Hear. Res. 78, 65-76).

Animals↗
PubMed1996

Active control of waves in a cochlear model with subpartitions.

Multiscale asymptotic methods developed previously to study macromechanical wave propagation in cochlear models are generalized here to include active control of a cochlear partition having three subpartitions, the basilar membrane, the reticular lamina, and the tectorial membrane. Activation of outer hair cells by stereocilia displacement and/or by lateral wall stretching result in a frequency-dependent force acting between the reticular lamina and basilar membrane. Wavelength-dependent fluid loads are estimated by using the unsteady Stokes' equations, except in the narrow gap between the tectorial membrane and reticular lamina, where lubrication theory is appropriate. The local wavenumber and subpartition amplitude ratios are determined from the zeroth order equations of motion. A solvability relation for the first order equations of motion determines the subpartition amplitudes. The main findings are as follows: The reticular lamina and tectorial membrane move in unison with essentially no squeezing of the gap; an active force level consistent with measurements on isolated outer hair cells can provide a 35-dB amplification and sharpening of subpartition waveforms by delaying dissipation and allowing a greater structural resonance to occur before the wave is cut off; however, previously postulated activity mechanisms for single partition models cannot achieve sharp enough tuning in subpartitioned models.

Animals↗
PubMed1996

Spontaneous firing of NG108-15 cells induced by transient exposure to ammonium chloride.

1. We report that NG108-15 (neuroblastoma x glioma) cells differentiated in defined serum-free media are capable of exhibiting stable automaticity (the spontaneous occurrence of regenerative action potentials) following exposure to extracellular perfusates containing NH4Cl. 2. Membrane depolarization (4-5 mV) concomitant with an increased pHi during NH4Cl exposure are followed by hyperpolarization (5-7 mV), sub-threshold oscillations, and spontaneous firing after the removal of NH4Cl. 3. Cells cultured in 10% serum did not exhibit automaticity. Cells cultured in serum-free media are twice as likely to show automaticity as those cultured in reduced (1.5%) serum media. 4. We have examined factors that contribute to the events following NH4Cl exposure, namely, membrane depolarization and hyperpolarization, subthreshold oscillations, and automaticity. The inward currents activated at more negative potentials and the ionic currents associated with pronounced afterhyperpolarization in NG108-15 cells cultured in serum-free media provide a basis for the repetitive activity in general and automaticity in particular.

Amiloride↗
PubMed1996

Ultrastructural localization of G-protein GS in the lateral wall of the guinea pig cochlear duct.

Immunocytochemical localization of a GTP-binding protein, Gs, in the various cells of the lateral wall of guinea pig cochlear duct was investigated using a post-embedding immunogold method with antibody raised against a synthetic decapeptide (RMHLRQYELL) encoding the C-terminus of the alpha-subunit of Gs. In the stria vascularis, labeling was observed on the basolateral membrane infoldings of marginal cells, on the juxtaposed membrane of intermediate cells, and on the cell membrane of basal cell. In contrast, no significant labeling was observed on the luminal membrane of marginal cells. Immunoreactivity also was detected on the cell membranes of various other cells. These include spiral prominence epithelial cells, fibrocytes of spiral ligament, external sulcus cells, and epithelial and mesothelial cells of Reissner's membrane. Adenylylcyclase has been functionally implicated in some of the cell types with membranes labeled in this study. The significance of these findings is briefly discussed.

Adenylyl Cyclases↗
PubMed1996

Viscoelastic relaxation in the membrane of the auditory outer hair cell.

The outer hair cell (OHC) in the mammalian ear has a unique membrane potential-dependent motility, which is considered to be important for frequency discrimination (tuning). The OHC motile mechanism is located at the cell membrane and is strongly influenced by its passive mechanical properties. To study the viscoelastic properties of OHCs, we exposed cells to a hypoosmotic solution for varying durations and then punctured them, to immediately release the osmotic stress. Using video records of the cells, we determined both the imposed strain and the strain after puncturing, when stress was reset to zero. The strain data were described by a simple rheological model consisting of two springs and a dashpot, and the fit to this model gave a time constant of 40 +/- 19 s for the relaxation (reduction) of tension during prolonged strain. For time scales much shorter or longer than this, we would expect essentially elastic behavior. This relaxation process affects the membrane tension of the cell, and because it has been shown that membrane tension has a modulatory role in the OHC's motility, this relaxation process could be part of an adaptation mechanism, with which the motility system of the OHC can adjust to changing conditions and maintain optimum membrane tension.

Animals↗