[Current role of electrocochleography].
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Acoustic Stimulation↗
Biomedical subjects
J M Aran
Publications and source records attributed to J M Aran.
Preparative purification of adenosine deaminase from human erythrocytes by affinity chromatography.
The purification of adenosine deaminase from human erythrocytes is reported. By means of classical procedures and by using affinity chromatography as the last step, the enzyme is purified 760,000-fold with a yield of 32%. The affinity resin is composed of purine riboside (nebularine) linked to Sepharose CL6B. Since the compound has no leaving group at the C-6 position the affinity gel is stable and the chromatography can be repeated several times (up to fifteen times in eight months). Purine riboside was chosen because its potency as a reversible inhibitor of adenosine deaminase is greater than that of inosine (a low-affinity inhibitor), but lower than that of erythro-9-(2-hydroxy-3-nonyl)adenine (a high-affinity inhibitor).
Adenosine Deaminase↗
Characterization of Na(+)-dependent, active nucleoside transport in rat and mouse peritoneal macrophages, a mouse macrophage cell line and normal rat kidney cells.
Peritoneal rat macrophages expressed solely an Na(+)-dependent, concentrative nucleoside transporter, which possesses a single Na(+)-binding site and transports purine nucleosides and uridine but not thymidine or deoxycytidine. The Michaelis-Menten constants for formycin B and Na+ were about 6 microns and 14 mM, respectively, and the estimated Na+:formycin B stoichiometry was 1:1. Rat macrophages accumulated 5 microM formycin B to a steady-state level exceeding that in the medium by about 500-fold during 60 min of incubation at 37 degrees C. Concentrative formycin B transport was resistant to inhibition by nitrobenzylthioinosine, lidoflazine, dilazep and nifedipine, but was slightly inhibited by high concentrations of dipyridamole (greater than 10 microM) and probenecid (greater than 100 microM). Mouse peritoneal macrophages and lines of mouse macrophages and normal rat kidney cells expressed Na(+)-dependent, active nucleoside transport but in addition significant Na(+)-independent, facilitated nucleoside transport. Facilitated nucleoside transport in these cells was sensitive to inhibition by nitrobenzylthioinosine, dilazep and dipyridamole. The presence of these inhibitors greatly enhanced the concentrative accumulation of formycin B by these cells by inhibiting the efflux via the facilitated transporter of the formycin B actively transported into the cells. Whereas rat macrophages lacked high-affinity nitrobenzylthioinosine-binding sites, mouse macrophages and normal rat kidney cells possessed about 10,000 such sites/cell. Rat and mouse erythrocytes, rat lymphocytes, and lines of Novikoff rat hepatoma cells, Chinese hamster ovary cells, Mus dunni cells and embryonic monkey kidney cells expressed only facilitated nucleoside transport.
Animals↗
Na(+)-dependent, active nucleoside transport in mouse spleen lymphocytes, leukemia cells, fibroblasts and macrophages, but not in equivalent human or pig cells; dipyridamole enhances nucleoside salvage by cells with both active and facilitated transport.
Formycin B influx studies have shown that P388 and L1210 mouse leukemia cells, mouse L929 cells, mouse RAW 309 Cr.1 cells, LK35.2 mouse B-cell hybridoma cells and cultured mouse peritoneal macrophages express both Na(+)-dependent, active and nonconcentrative, facilitated nucleoside transport systems. In the mouse cell lines, active transport represented only a minor nucleoside transport component and was detected only by measuring formycin B uptake in the presence of dipyridamole or nitrobenzylthioinosine, strong inhibitors of facilitated, but not of active, nucleoside transport. Inhibition of facilitated transport resulted in the concentrative accumulation of formycin B in cells expressing active nucleoside transport. Concentrative formycin B accumulation was abolished by treatment of the cells with gramicidin or absence of Na+ in the extracellular medium and strongly inhibited by ATP depletion or ouabain treatment. Mouse macrophages accumulated formycin B to 70-times the extracellular concentration in the absence of dipyridamole during 90 min of incubation at 37 degrees C. Thus active transport represents a major nucleoside transport system of these cells, similarly as previously reported for mouse spleen lymphocytes. In contrast to the various types of mouse cells, active formycin B transport was not detected in human HeLa cells, human H9, Jurkat and CEM T lymphoidal cells and pig spleen lymphocytes. These cells expressed only facilitated nucleoside transport with kinetic properties similar to those of the facilitated transporters of other mammalian cells.
Animals↗
Immunohistochemical evidence for GABAergic cell bodies in the medial nucleus of the trapezoid body and in the lateral vestibular nucleus in the guinea pig brainstem.
The presence of gamma-aminobutyric acid (GABA) in two brainstem nuclei is demonstrated by using a pre-embedding immunohistochemical procedure followed by staining intensification. Firstly, immunoreactivity was found in numerous cell bodies and profiles of the medial nucleus of the trapezoid body (MNTB). Secondly, numerous neurons including giant Deiters' cells, terminals and fibers were strongly labelled within the lateral vestibular nucleus (LVN). These observations suggest that the inhibitory part of the efferent innervation of outer hair cells in the cochlea can originate from the MNTB, and that GABAergic neurons in the LVN may contribute to information processing within this nucleus.
Animals↗
Na(+)-dependent, active and Na(+)-independent, facilitated transport of formycin B in mouse spleen lymphocytes.
Na(+)-dependent, active and Na(+)-independent facilitated nucleoside transport were characterized in mouse spleen cells using rapid kinetic techniques and formycin B, a metabolically inert analog of inosine, as substrate. The Michaelis-Menten constants for formycin B transport by the two transporters were about 30 and 400 microM, respectively. The first-order rate constant for Na(+)-dependent transport was about 4-times higher than that for facilitated formycin B transport. The Na(+)-dependent carrier is specific for uridine and purine nucleosides and accumulates formycin B concentratively in an unmodified form. Concentrative accumulation was inhibited by ATP depletion and gramicidin and ouabain treatment of the cells. Our data indicate a single Na(+)-binding site on the Na(+)-dependent nucleoside carrier and a Michaelis-Menten constant for Na+ of about 10 mM. This transporter was not significantly inhibited by dipyridamole and nitrobenzylthioinosine, inhibitors of the facilitated transporter. The Na(+)-independent, facilitated nucleoside transporter of spleen cells exhibits properties comparable to those of the carriers present in mammalian cells in general. The B lymphocytes remaining after depletion of spleen cell populations of T lymphocytes by incubation with a combination of T-cell specific monoclonal antibodies plus complement exhibited about the same activities of active and facilitated nucleoside transport as the original suspension.
Adenosine Triphosphate↗
Mobility of nucleoside transporter of human erythrocytes differs greatly when loaded with different nucleosides.
Time courses of transmembrane equilibration of 2-chloroadenosine, 2'-deoxyadenosine, 3'-deoxyadenosine, cytidine and 2'-deoxycytidine were measured by rapid kinetic techniques in human erythrocytes under equilibrium exchange and zero-trans conditions. The kinetic parameters for transport were computed by fitting appropriate integrated rate equations to the data pooled for seven concentrations and compared to the kinetic parameters for uridine, adenosine, thymidine and formycin B transport determined previously for human erythrocytes under comparable experimental conditions. The transport of all nucleosides conformed to the simple carrier model and was directionally symmetric. The Michaelis-Menten constants for equilibrium exchange (Kee) ranged from 22 microM for 2-chloroadenosine to about 4 mM for cytidine and the maximum velocities (Vee) differed in a similar manner, so that the first-order rate constants (Vee/Kee) were similar for all nucleosides. The kinetic parameters for 2'-deoxyadenosine transport were similar to those for adenosine transport, whereas the lack of the 3'-OH group greatly reduced the affinity of 3'-deoxyadenosine (cordycepin) for the carrier. 2', 3'-Dideoxynucleosides were transported less than 1% as efficiently as 2'- and 3'-deoxynucleosides. Thus, the 2'- and 3'-OH groups play an important role in nucleoside transport. The mobility of the carrier when loaded with pyrimidine nucleosides (reflected by Vee) was 5-10-times greater than that of the empty carrier, whereas the mobility of the adenosine-loaded or 2'-deoxyadenosine-loaded carrier was about equal to that of the empty carrier. Loading the carrier with 2-chloroadenosine or 3'-deoxyadenosine actually decreased its mobility. Thus, the differential mobility of the loaded and empty carrier differs greatly with the nucleoside substrate. The mobility of the loaded carrier as well as Kee increased with a decrease in lipid solubility of the nucleoside substrate, but the relationship was complex.
2-Chloroadenosine↗
Association of adenosine deaminase with erythrocyte and platelet plasma membrane: an immunological study using light and electron microscopy.
Adenosine deaminase has been localized in the plasma membrane of erythrocytes and platelets by means of immunological techniques using light and electron microscopy with cells in suspension. In erythrocytes, adenosine deaminase is associated with the external side of the plasma membrane. In platelets, the enzyme is associated with the external side of the plasma membrane, which is known to extend through the canalicular system of these cells. These results confirm our previous findings, based on biochemical studies, concerning the attachment of the enzyme to cell membranes.
Adenosine Deaminase↗
[Synergism between noise and ototoxic drugs: new experimental data].
Results of experiments investigating, in the guinea pig, the effects of combinations of gentamicin (GM), ethacrynic acid (EA) and acoustic stimulations are reported. They indicate that functional and morphological alterations develop only after GM has penetrated specifically into the sensory cells, that this penetration is indirectly facilitated by EA, since it is known to increase GM concentration in endolymph, and directly by functional depolarization of the cell, as it occurs during normal acoustic stimulation.
Animals↗
Electrovestibulography in experimental animals.
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Animals↗
Aminoglycoside antibiotics impair calcium entry but not viability and motility in isolated cochlear outer hair cells.
Cochlear outer hair cells have been well established as primary targets of the ototoxic actions of aminoglycoside antibiotics. These cells, isolated from the guinea pig cochlea and maintained in short-term culture, were used as a model for evaluating the acute effects of gentamicin on cell viability, depolarization-induced transmembrane calcium flux, and depolarization-induced motile responses. On the basis of morphology and fluorochromasia, the presence of extracellular gentamicin as high as 5 mM did not affect the viability of the cells for up to 6 hr, the longest time tested. Viable cells showed binding of fluorescently tagged gentamicin to their base but excluded the drug from their cytoplasm. In response to [K+]-depolarization, intracellular calcium levels (monitored with the fluorescent calcium-sensitive dye fluo-3) increased from a resting value of 218 +/- 102 nM to 2,018 +/- 1,077 nM concomitant with a cell shortening of 0.7% +/- 1.3%. The depolarization-induced calcium increase was apparently caused by calcium entry into the cell as it was inhibited by the calcium-channel blocker methoxyverapamil and prevented in the absence of extracellular calcium. Both gentamicin and neomycin blocked the [K+]-induced calcium increase at an IC50 of 50 microM. Despite the inhibition of calcium entry the ability of the outer hair cells to shorten under [K+]-depolarization was not impaired; in fact, cell shortening was even more pronounced in the absence of calcium influx (2.6% +/- 1.4%). This argues effectively against the existence of a calcium-dependent actomyosin-mediated component in [K+]-induced shape changes.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Limits of ABR and contribution of transtympanic electrocochleography in the assessment of cerebellopontine angle tumours.
Auditory brainstem responses (ABR) and transtympanic electrocochleography (ECochG) were analysed for 63 patients with tumours of the cerebellopontine angle (CPA) and/or internal auditory meatus (IAM). ABR recordings indicated a clearly prolonged wave I-V interval (above 4.3 ms) in half of the patients, hence suggesting a retrocochlear disorder. For the remaining patients ABR alone was not sufficient for clear diagnosis and ECochG was therefore used in addition. When the latency of wave V was uncertain due to the absence of wave I, the latency of wave N1 was always measurable by ECochG. The NI-V interval evaluated in this way was always significantly prolonged (above 4.5 ms) relative to a control group of patients with a sensorineural hearing loss. In 9 of the 13 subjects without any discernible ABR, an ECochG response could be recorded and its threshold was often better than the mean pure tone audiogram. This study confirms the validity of transtympanic ECochG in the diagnosis of retrocochlear disorders. The time difference observed between wave N1 (ECochG) and wave I (ABR) in CPA tumours is discussed and a possible explanation is proposed.
Audiometry, Evoked Response↗
Neuroanatomical study of Galen's anastomosis (nervus laryngeus) in the dog.
To further knowledge of the laryngeal nerves, the nerve fibers of Galen's anastomosis were studied using two neuroanatomical methods, namely nerve degeneration and horseradish peroxidase labeling. It is demonstrated that the superior laryngeal nerve forms part of the tracheal and esophageal nervous system. The value of the results in relation to physiological laryngeal studies and to human laryngeal diseases is discussed.
Animals↗
Effects of electrolytic lesions of the superior olivary complex and trapezoid body on brainstem auditory-evoked potentials in the guinea pig. I. Vertex-tragus recordings.
Brainstem auditory-evoked potentials (BAEP) were recorded between vertex and tragus in 22 guinea pigs, after destruction of the contralateral ear in order to produce monaural stimulation. Stimuli employed were 100 dB SPL clicks and tone bursts. Normative recordings showed five positive peaks, P1-P5 and four negative peaks, N1-N4. Electrolytic lesions were then made to the superior olivary complex (SOC) ipsilateral to the stimulation (6 animals), contralateral to the stimulation (8 animals), and trapezoid body (TB) (8 animals). In each group a statistical analysis of the results (amplitudes and latencies) was made (paired t test). After lesions to the ipsilateral SOC, there was an increase in the amplitude of P1 and a decrease in P3. After lesions to the contralateral SOC, there was a larger decrease in P3, which disappeared in 2 guinea pigs which had a large lesion involving many TB fibers. After TB lesions, there was a decrease in the P2-N2 composite wave, disappearance of P3, a decrease in P4, and an increase in the latency of P1 and P1-P2 and P2-P4 intervals. After ipsilateral SOC lesions, which certainly involved the uncrossed olivocochlear efferent tract, the increase in P1 suggested a disinhibition of the efferent fibers. The large decrease or the loss of P3 after TB lesions and SOC lesions involving many TB fibers suggested that P3 could be generated by the contralateral part of the TB fibers, in the vicinity of the contralateral SOC. These data are in agreement with the predominance of fiber tracts in the generation of BAEP.
Animals↗
Effects of electrolytic lesions of the superior olivary complex and trapezoid body on brainstem auditory-evoked potentials in the guinea pig. II. Three-channel Lissajous' trajectory analysis.
Brainstem auditory-evoked potentials (BAEP) were studied both with vertex (positive) - tragus (negative) derivation and three-channel Lissajous' trajectory (3-CLT) planar analysis in guinea pigs (GP) after destruction of the ipsilateral superior olivary complex (SOC) (6 GP), contralateral SOC (8 GP) referred to the stimulation side, and trapezoid body (TB) in the sagittal line (8 GP). The size of the lesion was evaluated on histological examination. The right ear was locally destroyed with aminoside instillation. The left ear was stimulated with unfiltered clicks and tone bursts of different frequencies at 100 dB SPL. Three questions were the basis of this study: (i) What are the effects of brainstem lesions on 3-CLT? (ii) Is there a relationship between planar segments and underlying sources? (iii) What is the generator of planar segment C (P3) in GP? 3-CLT analysis was made on plane A (corresponding to P1 on a vertex-tragus channel), plane B (N1-P2) and plane C (P3.) Plane C disappeared after TB lesions and remained after ipsi- and contralateral SOC destructions. However, plane C disappeared after extensive lesions to the contralateral SOC affecting the TB. Planes A and B were always observed whatever the lesion, with sometimes a modification of their orientation. A localized brainstem lesion induced a local modification of the three-dimensional trajectory. These data are in agreement with the assumption that some planes of 3-CLT reflect the activity of underlying generators. However, the loop which contains the planar segment is a better indicator of the global electrical activity of the source. Plane C (P3) would have a contralateral source which is probably not the SOC but the afferent curved fibers of the TB close to the contralateral SOC.
Animals↗
[Peripheral auditory adaptation in guinea pigs: comparison with loudness adaptation in man].
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Acoustic Stimulation↗
Clinical significance of the summating potential in Menière's disease.
Transtympanic (TT) electrocochleography (ECochG) data recorded with click stimuli and tone bursts (1, 2, 4, and 8 kHz) were evaluated in 50 patients with Meniere's disease and compared with the data from control groups of 10 sensorineural hearing impaired patients and 5 subjects with normal hearing. The mean summating potential (SP) amplitude was larger in the Meniere's disease group for 1, 2, and 8 kHz. The low frequency (1 or 2 kHz) SP decreased in 59% of the Meniere's disease patients during a glycerol dehydration test, whereas subjective hearing improved in only 29%. The use of ECochG with the monitoring of SP thus improves the sensitivity of the glycerol test for the detection of endolymphatic hydrops. There appeared no clear relation between clinical observations and the ECochG data either before or after glycerol. However, the Meniere's disease patients with a large negative SP at low frequencies also had larger action potential (AP) and often had short-term symptoms. The present report confirms the usefulness of measuring the SP in the diagnosis of Meniere's disease.
Action Potentials↗
Osmotically induced motility of outer hair cells: implications for Menière's disease.
Osmolarity changes in inner ear fluids have long been considered to be contributing factors to Menière's disease. Our present study demonstrates that small changes in the osmolarity of a surrounding in vitro medium induce fast contractions (hypo-osmotic solution) or elongations (hyperosmotic solution) in isolated outer hair cells of the guinea pig. These changes were reversible upon returning cells to iso-osmotic conditions. Up to five cycles of shape change could be sustained by these cells without obvious detriment to their morphology. These findings suggest that fluctuant changes in osmolarity of inner ear fluids can result in similar fluctuant changes of hair cell shape. Since the outer hair cells may control cochlear micromechanics and function by their active motility, osmotically induced abnormalities of cell dimensions and motility may contribute to the audiological manifestations of fluctuant hearing loss.
Animals↗