[Implication of mitochondrial apoptosis in neural degeneration of cochlea in a murine model for presbycusis]. / Implication de l'apoptose mitochondriale dans la dégénérescence neurale de la cochlée dans un modèle murin de presbyacousie.

HYPOTHESIS: Pathologies of senescence, in particular those of neurosensory organs represent an important health problem. The improvement of the life expectation entails the fast increase of the frequency of the presbyacusis in the population. The biological and molecular causes of this degenerative pathology of the inner ear are linked to the disappearance of the sensory cells (inner and outer hair cells) and are associated to nervous damages of the spiral ganglion in the cochlea. We were interested in mechanisms causing the cochlear degeneration in a model of mouse CD 1 presenting prematurely auditive losses. MATERIALS AND METHODS: We tried to correlate the evolution of the hearing and the appearance of apoptotic phenomena by marking with specific antibody, activated anti-caspase-3, in the cochlea during time. We studied the role and the involvement of proteins controlling the apoptosis as the P53 protein and from an energy point of view at the level of the mitochondria such as proteins of the Bcl-2 family and the cytochrome c in the various structures of the cochlea. RESULTS: After implantation of electrodes for auditory nerve acoustic thresholds measurements, the audition of mice CD 1 presented a characteristic profile of hearing losses which begins in the high frequencies from the age of 1 month and which quickly evolves towards the low frequencies. The observation (between the 1st and 3rd month of age) of spiral ganglion cells revealed an unchanged number of cellular bodies of type 1 neurons, on the other hand a characteristic morphology of apoptosis of glial cells with the formation of apoptotic body was noted. Indeed, glial cells expressed activated caspase-3. Furthermore, this phenomenon seems to be under the control of the pro-apoptotic protein Bax by its overexpression and a increased release of the cytochrome c. This phenomenon was followed at 3 and 6 months by the disappearance of the outer hair cells by 9 and 48% respectively. CONCLUSION: The apparition of the deafness in the murin model CD 1 allowed us to demonstrate that the degeneration of cochlear structure begins at the level of glial cells of the spiral ganglion from 3 months, followed thereafter by the deterioration of the nervous conduction between the spiral ganglion and the sensory cells. As a consequence, because of the impoverishment in nervous signals, the outer hair cells would begin to disappear during the 6th month. In conclusion, the understanding of the sequence and the cause of these mechanisms responsible for the neural degeneration and the loss of hearing could eventually, allow us to optimize the various treatments of the presbyacusis.

Sympathectomy improves the ear's resistance to acoustic trauma--could stress render the ear more sensitive?

Emotional stress is a phenomenon experienced by many people at some time in their lives. Some of its early manifestations, such as unbearable loudness of ambient sounds and sensations of dizziness, might be linked to inner ear dysfunction. Although the inner ear is supplied with a substantial sympathetic innervation, previous studies have failed to demonstrate any significant functional impact. We show here that in the awake guinea pig and following unilateral ablation of the superior cervical ganglion, the temporary threshold shift induced by a 1-min exposure to 8 kHz pure tone at 96 dB sound pressure level was reduced by as much as 40 dB. Of interest, the protective effect was bilateral suggesting an intimate relationship between the sympathetic and the olivocochlear efferent systems. The data presented here provide new evidence for a key role for the sympathetic system in modulating temporary threshold shifts following exposure to moderate sound stimulation. This opens new perspectives for investigation of sympathetic control in noise-induced permanent hearing losses.

Hyperbaric-induced enhancement of noradrenaline-evoked contraction in rat thoracic aorta.

The aim of this study was to investigate the effects of a high ambient pressure of He on vascular contraction induced by noradrenaline and to distinguish the effects of ambient pressure per se from those of increased pressure of inert gas. Rings of thoracic aorta were isolated from male Wistar rats. Isometric tension was measured in preparations exposed to 7.1 MPa (absolute pressure) of He. Dose-response curves for noradrenaline and contractions elicited by 120 mM KCl were compared with time-matched experiments performed at atmospheric pressure. The same protocol was also carried out under 7.1 MPa of N2. At the high pressure of He, the contraction elicited by noradrenaline was increased with no change in the response to K(+)-evoked depolarization. The tension developed in response to noradrenaline also increased under 7.1 MPa of N2 but the effects were less marked than during the He experiments. Moreover, the response to KCl was reduced in this circumstance. Hyperbaric conditions enhance the noradrenaline-induced contraction of rat aorta in vitro. This effect probably results from an action of pressure per se on activation of adrenoceptors. However, the hyperbaric-induced increase in vascular smooth muscle contraction is partially counteracted by high pressures of inert gases (N2, but also probably He), which impair the efficiency of the contractile machinery.

Relationship between inspired and expired gas temperatures in a hyperbaric environment.

When breathing room air at sea level the expired gas temperature (TE) increases in proportion to the inspired one (TI). Previous studies conducted under hyperbaric conditions have assumed that the TE vs TI relationship was the same when humans breathed room air at atmospheric pressure or helium-oxygen mixture under hyperbaric conditions. We hypothesized that the use of dilutant gases, as helium (He) or hydrogen (H2), having low density but high specific heat compared to nitrogen, could change the TE vs TI regression. The present study was conducted on 3 professional divers participating in the COMEX Hydra IX experiment. Three conditions were studied: A, (23.5 ATA, He-H2-O2 mixture); B, (21 ATA, H2-O2 mixture); and C, (21 ATA, He-O2 mixture). In each condition six different inspired temperatures were tested, while minute ventilation, TI and TE values were measured simultaneously. In all cases a linear relationship was found between TE and TI, but the slopes of the regression lines obtained in conditions A and B (gas mixture containing H2) were significantly lower than in condition C (He-O2 mixture). Computation of the convective respiratory heat loss (Cr) revealed that, when the subjects breathed the coldest gas mixtures (+10 degrees C), Cr value was 1.6 times higher in condition B than in C. These data are consistent with theoretical considerations and they demonstrate that a single equation cannot be used to predict the TE vs TI relationship in all environmental circumstances.

Respiratory effects of cold-gas breathing in humans under hyperbaric environment.

Changes in total lung resistance (RL) during inhalation of cold gas mixtures were measured in 4 human volunteers during an experimental dive at 46 ATA. The subjects breathed helium-nitrogen-oxygen mixtures during the decompression schedule, and measurements were performed at 46, 36, 21, 12.5, 6 and 2 ATA (1 ATA = 100 kPa). RL was measured during eupneic ventilation when individuals inhaled either ambient gas at +30 to +33 degrees C (control condition), or cooled gas at +7 to +18 degrees C. RL values measured in control conditions increased with gas density. Thus, the changes in RL induced by cold gas breathing were expressed in percent of the corresponding control values. No cold-induced bronchospasm occurred at low ambient pressure, even at the lowest inspired temperature, +7 degrees C. However, the airway response was present at pressure up to 21 ATA and then occurred at higher level of inspired gas temperature. The convective respiratory heat loss (Cr), calculated at each pressure level and experimental condition, was linearly related to cold-induced changes in RL; the value of Cr inducing 20% increase in RL was around 1.4 kcal.min-1. The bronchomotor response was related to the increase in respiratory heat loss induced by the high thermal capacity of the gas mixture used in hyperbaric environment. The present observations confirm previous data obtained under hyperbaric conditions (25 ATA) as well some experiments performed at sea level in normal individuals breathing very cold air.

Bronchomotor response to cold air or helium-oxygen at normal and high ambient pressures.

Effects of inhalation of cold air or helium-oxygen mixture on lung resistance (RL) were studied in anesthetized and tracheotomized rabbits under normal ambient pressure and in human volunteers under normo- and hyperbaric conditions. In artificially ventilated rabbits, an increase in RL occurred when the tracheal temperature fell to 10 degrees C. This effect was more than double with helium breathing compared to air, despite a lower respiratory heat loss by convection (Hc) with helium. In 3 normal humans, inhalation of cold air (mouth temperature = 8 degrees C) at sea level had no effect on RL value. However, with a helium-nitrogen-oxygen mixture, a weak but significant increase in RL due to cold gas breathing was measured in 1 subject at 2 ATA and in 2 individuals at 3.5 ATA. The density of inhaled gas mixture (air or He-N2-O2) was near the same in the three circumstances (1, 2, and 3.5 ATA) but Hc value increased with helium. At 8 ATA a 30-55% increase in RL occurred in the 3 divers during inhalation of cold gas (Hc was multiplied by 6 compared to air at sea level) and at 25 ATA the cold-induced bronchospasm ranged between 38 and 95% (Hc multiplied by 27). Thus, in rabbits and humans helium breathing enhanced the cold-induced increase in RL at normal or elevated ambient pressure, and this effect was interpreted as resulting from different mechanisms in the two circumstances.