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T Takasaka

Publications and source records attributed to T Takasaka.

At least 19 recordsLinked to original sources
PubMed1992

Effects of Ca2+ antagonists and aminoglycoside antibiotics on Ca2+ current in isolated outer hair cells of guinea pig cochlea.

The effects of various Ca2+ antagonists and aminoglycoside antibiotics on the Ca2+ channel in isolated outer hair cells of the guinea pig were investigated using a whole-cell patch-clamp technique. The inhibitory action was in the order of La3+ much greater than Cd2+ much greater than Ni2+ greater than Co2+ for inorganic Ca2+ antagonists, and flunarizine = nicardipine greater than omega-conotoxin greater than methoxyverapamil = diltiazem much greater than amiloride for organic ones. Aminoglycoside antibiotics also had antagonistic effects on the Ca2+ channel.

Aminoglycosides↗
PubMed1992

Glycine response in isolated dorsal cochlear nucleus of C57BL/6J mouse.

Pharmacological properties of glycine (Gly)-induced Cl- current (ICl) in the dorsal cochlear nucleus (DCN) neurons acutely dissociated from C57BL/6J mouse were investigated in the whole-cell configuration of the patch-clamp technique. Gly-induced ICl increased in a sigmoidal manner with higher Gly concentrations. Strychnine blocked the Gly response competitively at low and non-competitively at high concentrations. Both glutamate (Glu) and N-methyl-D-aspartate (NMDA) responses were augmented by adding 10(-6) M Gly, at which concentration Gly did not induce any ICl. This facilitation was not affected by strychnine. Our results clearly show the existence of strychnine-sensitive and -insensitive glycine receptors in the DCN neurons.

Animals↗
PubMed1992

Na(+)-Ca2+ exchange in the isolated cochlear outer hair cells of the guinea-pig studied by fluorescence image microscopy.

The outer hair cell isolated from the guinea-pig was superfused in vitro and the cytosolic calcium concentration ([Ca2+]i) and sodium concentration ([Na+]i) were measured using fluorescence indicators. Under the resting condition, [Ca2+]i and [Na+]i were 91 +/- 9 nM (n = 51) and 110 +/- 5 mM (n = 12), respectively. Removal of external Na+ by replacing with N-methyl-D-glucamine (NMDG+) increased [Ca2+]i by 270 +/- 79% (n = 27) and decreased [Na+]i by 23 +/- 4 mM (n = 6). Both changes in [Ca2+]i and [Na+]i were totally reversible on returning external Na+ to the initial value and were inhibited by addition of 0.1 mM La3+ or 100 microM amiloride 5-(N,N-dimethyl) hydrochloride. Elevation of external Ca2+ ions to 20 mM reversibly decreased [Na+]i by 8 +/- 6 mM (n = 5). Moreover, the chelation of the intracellular Ca2+ with 1,2-bis (2-aminophenoxy) ethane-N,N,N',N'-tetraacetic acid (BAPTA) exerted an inhibitory action on the NMDG(+)-induced reduction in [Na+]i. Exposure to 5 mM NaCN for 2 min significantly and reversibly increased [Ca2+]i by 290 +/- 37% (n = 5), but did not affect the [Ca2+]i elevation induced by the NMDG+ solution. The rise in [Ca2+]i induced by the NMDG+ solution was not enhanced by ouabain pretreatment. Addition of ouabain did not alter the [Na+]i. The present results are best explained by the presence of an Na(+)-Ca2+ exchanger in cell membrane and indicate that the activity of Na+/K+ pump is poor in outer hair cells.

Amiloride↗
PubMed1992

Quantitative carbohydrate analyses of the tectorial and otoconial membranes of the guinea pig.

Carbohydrate composition of the tectorial membrane (TM) and the otoconial membrane (OM) of the guinea pig was analyzed after hydrolysis, using high-performance anion-exchange chromatography and pulsed amperometric detection. Both of the tissues were highly glycosylated; the carbohydrate content being 24-42% of protein. GlcN, Gal, Glc and Man were found to be the major component sugars of TM, whereas little GalN was found. Fuc and NANA were also present, but NGNA was not detectable. After digestion with thermolysin for solubilization, OM was separated into two fractions: insoluble mineral particles of the otoconia (OM-ppt) and a soluble fraction from the gelatinous layer (OM-sup). These two fractions showed distinct carbohydrate composition from each other. Further analyses using glycosidases revealed that TM contained asialyl and monosialyl but little di-, tri- and tetrasialyl N-glycosides, and OM-sup did not seem to be susceptible to endo-beta-galactosidase, which is known to cleave some N-acetyl-polylactosamine and keratan sulfate. Based on these analyses, it can be suggested that most of the carbohydrates in TM are likely to be asialyl and monosialyl N-glycosides. N-Glycosides may be predominant in the otoconia as well, and a polymer structure consisting of GlcN(Ac) and Gal other than N-acetyl-polylactosamine may exist in the gelatinous layer of OM. O-Glycosylation of the usual type appeared to be minor in all the fractions.

Acetylation↗
PubMed1992

Membrane potential measurement in isolated outer hair cells of the guinea pig cochlea using conventional microelectrodes.

Membrane potential of the isolated outer hair cells (OHCs) from the guinea pig cochlea was measured using conventional microelectrodes filled with 200 mM KCl. The resting membrane potential during superfusion with the standard physiological saline solution containing 3.5 mM K+ was -47.3 +/- 1.4 mV (N = 72), which was higher than those previously reported for isolated OHCs studied by using microelectrodes. Addition of ouabain (10(-5)-10(-3) M), the specific Na+, K+ ATPase inhibitor, depolarized the cell slowly and progressively, indicating the presence of low but definite Na+, K+ ATPase activity in the plasma membrane of OHCs. The magnitude of membrane potential was mainly dependent on the extracellular K+ concentration ([K+]O). A ten-fold increase of [K+]O depolarized the membrane potential by 49.6 +/- 1.0 mV (N = 58). A decrease of [Na+]O to one tenth of the control hyperpolarized the membrane potential by about 2 mV. Decreasing extracellular Cl- from 131.3 mM to 27.5 mM did not cause a significant change in the membrane potential. Using the Goldman-Hodgkin-Katz equation, assuming a negligible contribution of Cl- to the membrane potential and total monovalent cat ion concentration of the cytosol similar to the extracellular fluid, we calculated the permeability ratio of K+ versus Na+ to 131 +/- 19 and intracellular K+ concentration to 33.3 +/- 1.9 mM.

Animals↗
PubMed1992

Intracellular pH regulation in isolated cochlear outer hair cells of the guinea-pig.

1. The intracellular pH (pHi) regulation mechanisms of the outer hair cell (OHC) isolated from the guinea-pig were studied using fluorescence ratio imaging microscopy. 2. The OHC pHi in the resting condition was 7.26 +/- 0.08 (mean +/- S.D., n = 49) when the standard solution buffered with HEPES-Tris was superfused. 3. Exposure to 25 mM-NH4+ in the absence of HCO3- caused biphasic changes in pHi; a transient increase (7.89 +/- 0.14, n = 22) followed by a slow decrease (7.57 +/- 0.12; mean +/- S.D.). Removal of external NH4+ by introducing the N-methyl-D-glucamine (NMDG+) solution in the absence of HCO3- markedly acidified the pHi to 6.38 +/- 0.12 with little pHi recovery. Subsequent application of the standard Na+ solution restored the pHi to the initial value. The recovery was inhibited by 0.5 mM-amiloride but not by 0.3 mM-DIDS (4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid). 4. In the presence of HCO3-, removal of both external NH4+ and Na+ promptly caused an intracellular acidification followed by a pHi recovery. The pHi recovery from an acid load was inhibited by 0.3 mM-DIDS or 10 microM-NPPB (5-nitro-2-(3-phenylpropyl-amino)-benzoate). However, the pHi in the steady state in the presence or absence of HCO3- was not altered by addition of 0.5 mM-amiloride or NMDG+ solution. 5. The intracellular buffering power obtained from the NH4+ exposure and withdrawal was -15.1 +/- 8.7 mM (pH unit)-1 (n = 6) and -14.3 +/- 5.8 mM (pH unit)-1, respectively. 6. Replacement of external Cl- with gluconate in the HCO3- solution increased the pHi from 7.22 +/- 0.12 to 7.51 +/- 0.20 (n = 6), which was inhibited by 0.3 mM-DIDS. Moreover, addition of DIDS to the HCO3- solution increased the pHi by 0.13 +/- 0.08 (n = 8). 7. When the external standard solution buffered with HEPES-Tris was replaced with the HCO3- solution, the basal pHi (7.27 +/- 0.10) was promptly acidified to 6.87 +/- 0.10 then relaxed slowly to 7.00 +/- 0.15 (n = 16). 8. The pHi showed an initial alkalinization and a subsequent slow acidification after the HCO3(-)-free standard solution replaced the HCO3- solution. The slow acidification was inhibited by low external Cl- concentration or by addition of 0.3 mM-DIDS.(ABSTRACT TRUNCATED AT 400 WORDS)

Acetazolamide↗
PubMed1992

Middle ear inflation with a gas mixture.

We studied the middle ear (ME) pressure in 13 atelectatic ears to evaluate the effect of politzerization or ME inflation by a catheter using both room air and a gas mixture. The gas contained 12% O2, 6% CO2 and 82% N2. Tympanometry was performed before and after the treatment. The elevated ME pressure induced by the treatments, whether with room air or the gas mixture, declined rapidly within the first 30 min. However, at 35 and 45 min after the treatment with room air, the mean decrease in pressure was significantly greater than with the gas mixture, which might be due to slow oxygen absorption through the ME mucosa.

Acoustic Impedance Tests↗
PubMed1992

MRL/MP-lpr/lpr mouse as a model of immune-induced sensorineural hearing loss.

Hearing acuity and inner ear disorders of MRL/lpr mice, bred for the study of autoimmune disease, were examined in comparison to those of BALB/c mice. The auditory brain stem response threshold of 20-week-old MRL/lpr mice was significantly higher than that of BALB/c mice of the same age (p < .01). The pathologic changes of 20-week-old MRL/lpr mice were characterized by the degeneration of intermediate cells, widened intercellular spaces, and immunoglobulin G deposition on the basement membrane of strial blood vessels as well as in the basal infolding of strial marginal cells, which were absent in BALB/c mice. That there were no other evident pathologic findings in the cochlea or middle ear suggests that these changes in the stria vascularis seemed to be responsible for the sensorineural hearing loss of this mouse. The MRL/lpr mouse was thought to be a good experimental model to study the spontaneous sensorineural hearing loss caused by an immune reaction.

Animals↗
PubMed1992

[A case of Bezold's abscess associated with cholesteatoma].

Since the advent of antibiotics, otogenic complications have decreased considerably. However, incomplete antibiotic therapy has altered the clinical course of middle ear disease so as to be more insidious. This paper reports a case of Bezold's abscess associated with cholesteatoma. A 48-year-old man visited our hospital presenting with a 4-day history of right otorrhea and a tender swelling in the right neck. Physical examination showed a febrile patient (38.8 degrees C) with right facial paresis and trismus. A hyperemic, hard and tender swelling was observed in his right neck from the lateral cervical to the mental region. The tympanic membrane was invisible because of granulation and swelling of the posterior wall of the external auditory canal. Intravenous clindamycin and ceftazidime therapy was started immediately. A CT-scan revealed a diffuse shadow with bony destruction in the right mastoid cortex. Extensive abscess formation was also found in the right sternocleidomastoid muscle, in the anterior neck and in the posterior neck. He was diagnosed as having Bezold's abscess associated with cholesteatoma. Radical mastoidectomy and drainage of the neck abscess was performed on the third day under general anesthesia. The mastoid cavity was found to be filled with pus and cholesteatoma debris. A small area of defective bone was found at the mastoid tip, through which there were communications between the mastoid cavity and the abscesses in the neck. Bony destruction was also found in the horizontal and vertical portion of the facial canal. Bacteroides and three kinds of gram-negative rods were cultured from the mastoid cavity.(ABSTRACT TRUNCATED AT 250 WORDS)

Abscess↗
PubMed1992

[Evaluation of allergen-specific IgE antibody and total IgE with a new IgE detection system named FAST: fluorescence allergosorbent test].

The measurement of IgE antibodies and total IgE was performed with FAST (fluorescene allergosrobent test) system using 171 serum samples obtained from normal donors and 789 serum samples obtained from patients with various allergic diseases. These were collected by departments of internal medicine, pediatrics, otorhinolaryngology and dermatology at 17 institutes in Japan. In addition to a comparative study with RAST, skin tests and provocation tests were also performed to establish a clinical diagnosis. Simultaneous measurements were made and an excellent correlation with RAST was observed with a concordance rate of 85.7% and correlation coefficient of 0.848. The specificity determined in the normal serum samples was 95.5% by RAST and 93.1% by FAST. The sensitivity determined in the samples, of which etiological allergens were identified clinically, was 82.3% by RAST and 85.3% by FAST. Among them, the sensitivity of skin test and provocation test were 92.4% and 94.7%, respectively. The concordance rate with clinical diagnosis was as high as 89.7% for both RAST and FAST. The normal upper limit of total IgE by cumulative 95% value was calculated to be 250 IU/ml for adult subjects. These results indicated the clinical usefulness of the FAST system in evaluating IgE antibodies and total IgE.

Allergens↗
PubMed1991

Calcium channel in isolated outer hair cells of guinea pig cochlea.

The physiological and pharmacological properties of the Ca2+ channel in outer hair cells (OHCs) freshly isolated from guinea pig cochlea were investigated using a whole-cell patch-clamp technique. The Ca2+ current (ICa) was activated from a membrane potential of -20 mV and reached peak value around +20 mV in external solution containing 20 mM Ca2+ at a holding potential of -70 mV. The peak amplitude of ICa increased in a hyperbolic manner with increasing extracellular Ca2+ concentration. The ion selectively was Ba2+ much greater than Ca2+ greater than or equal to Sr2+. It was concluded that the Ca2+ channel in OHCs of guinea pig is of the L-type.

Action Potentials↗
PubMed1991

Effect on cochlear potentials of lateral semicircular canal destruction.

Recording of the cochlear potentials was successfully performed during experimental labyrinthectomy in the guinea pig and in three patients with acoustic neuromas during translabyrinthine removal of the tumors. In the guinea pig, complete interruption of the duct of the lateral semicircular canal including the endolymphatic canal caused little change in the endocochlear DC potential of the first cochlear turn and input-output function curve of the N1 component of the compound action potential elicited by 8-kHz tone bursts. Further drilling of the vestibular labyrinth in the guinea pig caused decline of these potentials when the vestibular was opened. In patients with acoustic neuromas, the interruption of the duct of the lateral semicircular canal hardly altered the N1 input-output function curve and N1 input-latency function curve during the 1-hour observation period. Consistent preservation of cochlear function even after interruption of lateral semicircular canals suggests the possibility of partial surgical labryrinthectomy with preservation of hearing for lesions involving semicircular canals.

Acoustic Stimulation↗
PubMed1991

Spontaneous otoacoustic emissions in the guinea pig.

Measurement of spontaneous otoacoustic emission (SOAE) in a large number of animals was conducted in the guinea pig. A remarkably high incidence of SOAE (51 ears out of 248 ears; 20.6%) was observed in the animals that were raised under unexceptional conditions. The frequencies of SOAE were distributed between 0.7 kHz and 2.3 kHz (average: 1.16 kHz). The emission signals disappeared with hypoxia and with furosemide injection, and recovered after certain periods with some frequency fluctuations. Their electrical correlates that behaved in exactly the same manner as SOAE were clearly distinguished by FFT analysis of the AC potential from the cochlear surface. The signals could be divided into 4 types: (1) emission with pure and stable frequency spectrum and steady level, (2) emission with stable frequency and fluctuating level, (3) emission alternating between two closely situated frequency points, (4) emission having an irregular, wide spectrum band. Light microscopy revealed no abnormalities in the organ of Corti that have been widely accepted to be SOAE-related, i.e., patchy lesions of the organ of Corti and/or irregularities in hair cell arrangement. This suggests that the spontaneous oscillation within the guinea pig cochlea can be generated by some minor, functional, and probably reversible disturbances of the active cochlear transduction mechanism, without major structural derangement of the organ of Corti.

Acoustics↗
PubMed1991

Effects of pH on intracellular calcium levels in isolated cochlear outer hair cells of guinea pigs.

Effects of intracellular pH (pHi) on intracellular Ca2+ (Cai2+) in the outer hair cell (OHC) were investigated using fluorescence ratio imaging microscopy. Cai2+ and pHi were determined with fluorescence indicators fura-2 and 2,7-bis(2-carboxyethyl)- 5,6-carboxyfluorescein, respectively. Intracellular alkalinization from the basal pHi of 7.22 +/- 0.03 to 7.82 +/- 0.09 (n = 7) induced by 25 mM NH4+ caused a rise in Cai2+ from 102 +/- 15 to 195 +/- 28 nM (n = 8). The elevation of Cai2+ was inhibited by removing external Ca2+ or by 50 microM nifedipine. On the other hand, the cytosolic acidification produced by the removal of NH4+ (delta pHi = 0.53 +/- 0.06, n = 7) and the admission of 5% CO2 (delta pHi = 0.72 +/- 0.05, n = 4) elicited a slight reduction in Cai2+. The depolarization of the membrane potential by exposure to 100 mM K+ induced an increase in Cai2+ that was susceptible to nifedipine, indicating the presence of voltage-sensitive Ca2+ channels. The elevation in Cai2+ induced by 100 mM K(+)-containing solution was increased by 138 +/- 6% (n = 4) by external alkalinization to pH 8.4 and was decreased by 87 +/- 3% (n = 4) by external acidification to pH 6.4. These results demonstrate that both internal and external alkalinization of OHCs facilitate the influx of Ca2+ through the Ca2+ channel, leading to a rise of Cai2+. Thus pH, extra- and intracellular, may modulate the OHC motility by regulating the Cai2+.

Amiloride↗
PubMed1991

Effect of neuroregulators on the intracellular calcium level in the outer hair cell isolated from the guinea pig.

The cytosolic calcium concentration [( Ca2+]i) of the isolated outer hair cell of the guinea pig was measured using fluorescence imaging microscopy and the effects of efferent neuroregulators such as acetylcholine, ATP, GABA, substance P, enkephalin, calcitonin gene-related peptide, serotonin, dopamine, norepinephrine, and glutamate were investigated. Among the drugs tested only ATP induced an elevation of the [Ca2+]i of the outer hair cell. In the resting condition, [Ca2+]i averaged 104.5 +/- 31.1 nM (n = 27), while 100 microM ATP significantly increased [Ca2+]i to 146.3 +/- 43.5 nM (n = 19). Superfusion with Ca2(+)-free solution (pCa = 7.5) abolished the increase in [Ca2+]i induced by ATP, suggesting that ATP causes an entry of external Ca2+. The relevance of [Ca2+]i to the inhibitory actions of efferent neuroregulators is discussed.

Acetylcholine↗
PubMed1991

Otic preparations altered permeability and thickness of the round window membrane of the chinchilla.

The effects of topical otic preparations (Cortisporin, Coly-Mycin, Aristocort, and Bestron) upon the permeability of the round window membrane (RWM) in chinchillas were investigated. Using K(+)-selective microelectrodes, the concentration of tetraethylammonium (TEA) ions was measured. Changes in the thickness of the RWM were measured using light microscopy. The RWM permeability was reduced significantly in Cortisporin- and Coly-Mycin-treated ears. Moreover, these two drugs resulted in a marked thickening of the RWM. In contrast, Aristocort or Bestron resulted in no alteration of the RWM permeability.

Animals↗
PubMed1991

Bone destruction mechanisms in chronic otitis media with cholesteatoma: specific production by cholesteatoma tissue in culture of bone-resorbing activity attributable to interleukin-1 alpha.

To clarify specific mechanisms underlying cholesteatoma-induced bone destruction, surgical specimens of middle ear inflammatory granulation tissue with or without cholesteatoma were maintained in vitro and the bone-resorbing activity in their culture supernatants was analyzed by means of calcium release from mouse calvaria. Almost the same levels of bone-resorbing activity and prostaglandin (PG) E2 were found in the supernatants of both types of tissue. By contrast, aural polyp tissue yielded hardly any such activity or PGE2. Under the influence of indomethacin, however, only tissue with cholesteatoma produced considerable bone resorption activity, whereas PGE2 production was suppressed completely. Such activity in the cholesteatoma culture supernatant was not due to contamination of endotoxin and proved to be blocked by the introduction of anti-interleukin (IL)-1 alpha antibody into the calvarial assay system. Anti-IL-1 beta antibody had no effect on such activity. Interleukin-1 alpha was detected only in cholesteatoma tissue culture supernatants by means of enzyme-linked immunosorbent assay and by bioassay. These data suggest that the bone destruction in otitis media with cholesteatoma may be attributed to IL-1 alpha in addition to PGE2.

Animals↗