Ikuo Kitano

Involvement of Cl- transport in forskolin-induced elevation of endocochlear potential.

To determine the possible involvement of Cl- transport in the forskolin-induced endocochlear potential (EP) elevation, the effect of forskolin on the EP was examined in Cl(-)-free artificial perilymph (aPL) as well as in the presence of Cl- channel blockers. The perfusion of scala vestibuli (SV) with forskolin (200 microM) dissolved in Cl(-)-free aPL failed to produce an EP elevation, while SV perfusion of forskolin dissolved in normal aPL elevated the EP. The application of DPC and IAA-94 (blockers of cAMP-activated Cl- channel) into SV completely suppressed the forskolin-induced EP elevation, while niflumic acid (a Ca(2+)-activated Cl- channel blocker) failed to do so. IAA-94 applied into scala tympani (ST) did not suppress this EP elevation. The results suggest that adenylate cyclase may modulate the EP by changing Cl- transport between SV and scala media (SM) across Reissners membrane.

Role of endolymphatic anion transport in forskolin-induced Cl− activity increase of scala media

To determine the role of anion transport in the forskolin-induced Cl- increase of scala media (SM), effects of forskolin on the EP (endocochlear potential) and Cl- activity (ACl) in SM were examined with double-barrelled Cl(-)-selective microelectrodes. The experiments were carried out on guinea pig cochleae, using a few anion transport inhibitors: IAA-94 for a Cl- channel blocker, bumetanide (BU) for an Na+/K+/2Cl- cotransport blocker, and SITS and DIDS for Cl-/HCO3- exchange blockers. The application of forskolin (200 microM) into scala vestibuli (SV) caused a 20 mEq increase of endolymphatic ACl and a 15 mV elevation of EP, and IAA-94 with forskolin completely abolished these responses. Although each application of BU, SITS or DIDS did not completely suppress EP elevation, the concurrent application of these inhibitors completely suppressed EP with endolymphatic ACl increase. The results indicate the involvement of Cl- channels, Na+/K+/2Cl- cotransport and Cl-/HCO3- exchange in forskolin-induced increase of ACl and EP. The role of adenylate cyclase activation and Cl- transport in endolymph homeostasis was discussed.

The effect of protein kinase C stimulator and inhibitor on cochlear potentials in the guinea pig

To determine a possible role of protein kinase C (PKC) in the cochlear, the effects of a PKC stimulator (phorbol-12-myristate-13-acetate; PMA), an inactive analogue of PKC stimulator (4 alpha-phorbol-12,13-didecanoate; 4 alpha-PDD) and a PKC inhibitor (D-sphingosine) on cochlear potentials were examined in the guinea pig. The perilymphatic perfusion with PMA (3 x 10(-6) M) produced an increase in compound action potential (CAP) amplitude and no change in N1 latency, the amplitudes of negative summating potential (-SP), cochlear microphonics (CM) and endocochlear potential (EP). The perfusion with 4 alpha-PDD (3 x 10(-6) M) did not change the sound-evoked cochlear potentials and the EP. The perfusion with D-sphingosine (10(-5) M) produced a decrease in CAP amplitude and no change in N1 latency and the amplitudes of -SP, CM and EP. The results suggest that PKC may be involved in the mechanism underlying the CAP generation.

Failure of forskolin to elevate the endocochlear potential in kanamycin-poisoned animals.

The effect of forskolin (FSK) on the endocochlear potential (EP), K+ activity (AK), Na+ activity (ANa) and Cl- activity (ACl) in scala media (SM) was compared between normal and kanamycin (KM)-poisoned guinea pigs by means of double-barrelled ion-selective microelectrodes. The perfusion of the scala vestibuli (SV) with FSK (200 microM) produced EP elevation in normal animals whereas FSK failed to do it in KM-poisoned animals. FSK increased ACl of SM with no significant change in AK and ANa of SM in both groups of animals. Histological examination of KM-poisoned animals showed damaged outer and inner hair cells with an intact appearance of the stria vascularis. The mechanism underlying the failure of FSK to elevate the EP in KM-poisoned animals is discussed.

The action of substance P methyl ester on cochlear potentials in the guinea pig

The action of the substance P agonist, substance P methyl ester (SPME) on cochlear potentials was examined in the guinea pig. Previous studies have shown that SPME is a selective agonist for neurokinin 1(NKI) receptor. Perfusion with SPME at a concentration of more than 10-6M produced an increase in the amplitudes of the compound action potential and negative summating potential in a dose-dependent manner. N1 latency showed a tendency to be shortened, but this change was not significant. Amplitudes of the cochlear microphonics and endocochlear potential remained unchanged. Substance P fragment 7–11, an inactive analogue, produced no changes in the cochlear potentials. In contrast, the substance P antagonist [D-Pro2,D-Trp7,9]-SP blocked the action of SPME on the cochlear potentials. These results suggest that substance P may modulate neurotransmission through NK1 receptors in the cochlea.

The effect of adenosine on cochlear potentials in the guinea pig

The effect of adenosine on cochlear potentials was examined in the guinea pig. Perilymphatic perfusion with 10−4 M adenosine produced a significant decrease in the amplitudes of cochlear microphonics, negative summating potential (-SP) and compound action potential (CAP) and significant prolongation of N1 latency with no change in the endocochlear potential. The decreases in the amplitudes of -SP and CAP caused by adenosine were dose-dependent. Perilymphatic perfusion with an inactive analogue, 8-bromoadenosine, produced no changes in the cochlear potentials. The A1-receptor agonist, 2-chloroadenosine, produced a similar change in cochlear potentials to adenosine, while no changes were produced by the A2-receptor agonist ,5′-(N-ethylcarboxamido)-adenosine. These results suggest that adenosine may have a modulatory function through an Al receptor in the cochlea.

Effect of Endoplasmic Ca2+-ATPase Inhibitors on Cochlear Potentials in the Guinea-pig

The effect of endoplasmic Ca2+-ATPase inhibitors on cochlear potentials was examined in the guinea-pig. Perilymphatic perfusion with thapsigargin (10[-6] M) produced a significant decrease in the amplitudes of cochlear microphonics, negative summating potential and compound action potential, and a significant prolongation of N1 latency with no change in the endocochlear potential. These changes were all dose dependent. Another endoplasmic Ca2+-ATPase inhibitor, cyclopiazonic acid (10[-5] M), produced the same effects as thapsigargin on cochlear potentials. These results suggest that endoplasmic Ca2+-ATPase inhibitors may have inhibitory functions on cochlear potentials.

The Effect of Forskolin on the Sound-evoked Potentials in the Guinea Pig Cochlea

The effect of forskolin (FSK) on cochlear sound-evoked potentials was examined in the guinea pig. The perfusion of the scala vestibuli (SV) with FSK (2 x 10(-4) M) produced a significant increase in the amplitude of negative summating potential (- SP) with no change in cochlear microphonics (CM) amplitude, and a significant decrease in the amplitude of compound action potential (CAP) with a significant prolongation of N1 latency and a 20 dB CAP threshold elevation. The results lead us to speculate that FSK-induced changes may be involved in the transient formation of endolymphatic hydrops.

Role of ATP-sensitive K+ channels in anoxia-sensitive negative potential of endolymph

To investigate the possible role of the ATP-sensitive K+ channels (KATP channels) in the generation of the anoxia-sensitive negative potential (ASNP), the effects of the treatment with glybenclamide and diazoxide on the endocochlear potential (EP) and K+ activity (AK) in the scala media were examined with double-barrelled K(+)-selective microelectrodes. The experiments were carried out in guinea pig cochleae, using glybenclamide as a KATP channel blocker and diazoxide as a KATP channel opener. Perilymphatic perfusion of glybenclamide decreased the amplitude of the ASNP and shortened its duration, whereas perfusion of diazoxide increased ASNP amplitude and prolonged its duration. Glybenclamide enhanced the decrease of endolymphatic AK by anoxia, whereas diazoxide suppressed this AK decrease. The results suggest that KATP channels may be involved in the generation of the ASNP.

Failure of forskolin to elevate the endocochlear potential in experimental endolymphatic hydrops of the guinea pig

The effect of forskolin (FSK) on the endocochlear potential (EP) in scala media (SM) was examined in experimental endolymphatic hydrops of the guinea pig. Two weeks after obliteration of the endolymphatic sac the EP of hydroptic ears and that of the contralateral control ears were measured by means of microelectrodes. The perfusion of scala vestibuli (SV) with FSK (200 microM) produced EP elevation in the contralateral control ears but failed to do so in the experimental hydroptic ears. Histological examination of experimental endolymphatic hydrops showed mild hydrops with intact appearance of outer and inner hair cells, and the stria vascularis. The mechanism underlying the failure of FSK to elevate the EP in experimental endolymphatic hydrops is discussed.