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K H Iwasa

Publications and source records attributed to K H Iwasa.

31 records · Page 2Linked to original sources
PubMed1995

Ultrastructural localization of G-protein Gs in the organ of Corti.

Immunocytochemical localization of a stimulatory GTP-binding protein Gs in the organ of Corti in the inner ear was examined with a post-embedding immunogold technique, using antibodies raised against a synthetic decapeptide (RMHLRQYELL) of the C-terminus of the alpha subunit of Gs. Immunoreactivity was strong on the membranes of supporting cells in the reticular lamina, including inner and outer pillar cells and the phalangeal process of Deiters' cells. Immunolabeling also was seen on the membranes of cell bodies of those cells which surround nerve fibers, basilar fibers, outer spiral fibers and afferent nerve endings at outer hair cells. Gold particles also labeled the membrane of inner phalangeal cells and border cells. In contrast, outer and inner hair cells were not labeled. Possible roles of Gs in the organ of Corti are discussed.

Amino Acid Sequence↗
PubMed1995

Amiloride-sensitive Na+ channel-like immunoreactivity in the luminal membrane of some non-sensory epithelia of the inner ear.

Some non-sensory epithelia of the inner ear were examined for the localization of immunoreactivity to polyclonal antibodies raised against amiloride-sensitive Na+ channels from the bovine kidney. The pre-embedding immunogold technique was used for this purpose. Labelings were found on the membrane of the endolymphatic surface of strial marginal cells, epithelial cells of spiral prominence and Reissner's membrane, and ampullar dark cells. In contrast, no labeling was found on the luminal membrane of mesothelial cells of Reissner's membrane, the cells lining the supra-strial perilymphatic space, transitional cells and ampullar ceiling cells. Since the antibodies used may also label non-selective cation channels and non-functional sodium channel precursors as suggested by others, it was not possible to determine the labelings are solely due to amiloride-sensitive Na+ channels. However, the observed result was consistent with the previous studies that amiloride blocks ion transport in strial marginal cells and the semicircular canal. It is therefore likely that the observed labeling includes amiloride-sensitive Na+ channels. These labeled ion channels in a variety of epithelial cells lining the endolymphatic space could be important in the inner ear fluid regulation.

Amiloride↗
PubMed1994

Amiloride-sensitive channels in marginal cells in the stria vascularis of the guinea pig cochlea.

We examined marginal cells in stria vascularis for the presence of amiloride-sensitive Na+ channels, a possible pathway for maintaining a low Na+ concentration in the endolymph. Whole-cell voltage-clamp experiment shows that amiloride at 1 microM concentration reversibly reduces inward current more than outward current. Immunogold-labeling method shows that the luminal and lateral membrane have antigenic sites for these antibodies. These observations indicate the presence of amiloride-sensitive channels in the marginal cell. If amiloride-sensitive channels in the luminal membrane are highly selective to Na+, they could be an efficient pathway for Na+ uptake from the endolymph. In the basolateral membrane, amiloride-sensitive Na+ channels may make a relatively small contribution to the unusual resting potential.

Amiloride↗
PubMed1994

A membrane motor model for the fast motility of the outer hair cell.

A model is presented for describing the membrane potential-dependent motility of the outer hair cell. This model assumes that the motility is due to conformational changes of motor molecules in the plasma membrane. Two kinds of experimental observations, elasticity of the cell and stretch dependence of the motor molecule, are important for characterize this motility by the model. The motor molecule can be described by a two-state model which has electrical and mechanical components in the free energy. The electrical component is due to the charge transferred across the membrane and the mechanical component is due to a change in membrane area in the two states. It can be shown that the elastic element and the motor element are connected in series. Thus the apparent strain of the cell is represented by the sum of true elastic strain and changes due to motor molecules. This model predicts the amplitude of the movement and the force produced by the motility. The model predicts the force produced under isometric condition is about 0.1 nN/mV, in agreement with values estimated from in vivo conditions. The effect of an elastic load attached to the cell is also discussed.

Cell Membrane↗
PubMed1993

Effect of stress on the membrane capacitance of the auditory outer hair cell.

The membrane capacitance of the outer hair cell, which has unique membrane potential-dependent motility, was monitored during application of membrane tension. It was found that the membrane capacitance of the cell decreased when stress was applied to the membrane. This result is the opposite of stretching the lipid bilayer in the plasma membrane. It thus indicates the importance of some other capacitance component that decreases on stretching. It has been known that charge movement across the membrane can appear to be a nonlinear capacitance. If membrane stress at the resting potential restricts the movement of the charge associated with force generation, the nonlinear capacitance will decrease. Furthermore, less capacitance reduction by membrane stretching is expected when the membrane is already extended by the (hyperpolarizing) membrane potential. Indeed, it was found that at hyperpolarized potentials, the reduction of the membrane capacitance due to stretching is less. The capacitance change can be described by a two state model of a force-producing unit in which the free energy difference between the contracted and stretched states has both electrical and mechanical components. From the measured change in capacitance, the estimated difference in the membrane area of the unit between the two states is about 2 nm2.

Animals↗
PubMed1992

A membrane-based force generation mechanism in auditory sensory cells.

Auditory outer hair cells can elongate and shorten at acoustic frequencies in response to changes of plasma membrane potential. We show that this fast bidirectional contractile activity consists of an electromechanical transduction process that occurs at the lateral plasma membrane and can be activated and analyzed independently in small membrane patches inside a patch electrode. Bidirectional forces are generated by increases and decreases in membrane area in response to hyperpolarization and depolarization, respectively. We suggest that the force generation mechanism is driven by voltage-dependent conformational changes within a dense array of large transmembrane proteins associated with the site of electromechanical transduction.

Animals↗
PubMed1992

Elasticity and active force generation of cochlear outer hair cells.

The cochlear outer hair cell is described by a cylindrical membrane model, characterized by area and shear moduli for a passive elastic element and an active tension element dependent on the membrane potential. In passive experiments, these moduli are determined from the pressure-strain relations. The area modulus obtained is 0.07 N m-1, similar to a lipid bilayer and the shear modulus is 0.007 N m-1. These moduli combined with previous active experiments show that the active tension is nearly isotropic and is about 1.6 x 10(-2) N m-1 V-1, resulting in a 0.5 nN/mV force per cell. This implies that the receptor potential for acoustical stimulation produces an active force comparable to the acoustic force applied to the basilar membrane per outer hair cell. This finding supports the hypothesis that the outer hair cell acts as feedback motor in the fine tuning mechanism of the mammalian ear.

Animals↗
PubMed1991

Stretch sensitivity of the lateral wall of the auditory outer hair cell from the guinea pig.

The inner and outer hair cells of the mammalian hearing organ are mechano-transducer cells. Here we report evidence that the lateral wall of outer hair cells (OHCs) is a mechano-receptor. This mechano-sensitivity appears to complement that of the stereocilia. Patch clamping studies showed that stretching of the membrane patches by suction at the pipette activated potassium channels with 130 pS unit conductance specifically localized in the lateral wall. Application of an osmotic tension to the entire cell membrane under whole-cell recording produced a 10 mV hyperpolarization. The reversal potential and the magnitude of the macroscopic current under voltage clamp were consistent with the single-channel properties of stretch-activated potassium channels. The elongated cylindrical cell body of the OHC is optimally positioned in the cochlea to sense axial force due to the vibrations of the basilar membrane during sound stimulation. This sensitivity can explain the production of a predominantly hyperpolarizing response to sound stimuli, unique to the OHC. Coupled with voltage-dependent OHC motility, the stretch-activated channels may play an important role in producing a mechanical feedback, an indispensable element in cochlear tuning.

Animals↗
PubMed1990

High concentration of inositol 1,4,5-trisphosphate in sea urchin sperm.

We measured inositol 1,4,5-trisphosphate (InsP3) content of sea urchin gametes by using a specific protein binding assay, and found that a spermatozoon contains 4 x 10(-19) to 1 x 10(-18) moles of InsP3 before the acrosome reaction. Since the acrosome reaction has previously been shown to increase the InsP3 content of sperm severalfold, our measurement indicates that a spermatozoon contains at least 2 x 10(-18) moles of InsP3 at fertilization, corresponding to a concentration in the spermatozoon of about 1 mM. The threshold for activation of eggs by injection of InsP3 dissolved in a much larger volume of solution has been found to be about 3 x 10(-18) moles, corresponding to a concentration in the injectate of 1 microM. This suggests that sea urchin sperm may contain enough InsP3 to activate eggs. With an electroporation method, we also showed that sperm extract acts on eggs only from inside, consistent with a primary messenger role for InsP3.

Acrosome↗
PubMed1989

Fast in vitro movement of outer hair cells in an external electric field: effect of digitonin, a membrane permeabilizing agent.

Isolated outer hair cells from the organ of Corti show elongation and contraction in response to an externally applied ac electric field as well as to a direct current injection into these cells. This is thought to be the basis of the positive feedback mechanism for fine tuning of the mammalian hearing organ. To test whether the mechanical response depends on the intracellular electric field or on the membrane potential, we used digitonin to shunt the membrane resistance. We observed that the application of digitonin abolished the cellular response of the outer hair cells to an ac external electric field (5-30 Hz). Coinciding with the abolition of the cellular response, the nuclear matrix started to oscillate synchronous to the external field, indicating an appreciable increase of the intracellular electric field. If the intracellular electric field was the regulating factor of the motile response, the initiation of the movement of the nuclear matrix would have been accompanied by an enhancement of the cellular movement. Our observation is therefore consistent with the interpretation that the (local) membrane potential, and not the intracellular electric field, regulates the hair cell movement.

Cell Membrane Permeability↗