28 research outputs found
In Vivo Outer Hair Cell Length Changes Expose the Active Process in the Cochlea
BACKGROUND: Mammalian hearing is refined by amplification of the sound-evoked vibration of the cochlear partition. This amplification is at least partly due to forces produced by protein motors residing in the cylindrical body of the outer hair cell. To transmit power to the cochlear partition, it is required that the outer hair cells dynamically change their length, in addition to generating force. These length changes, which have not previously been measured in vivo, must be correctly timed with the acoustic stimulus to produce amplification. METHODOLOGY/PRINCIPAL FINDINGS: Using in vivo optical coherence tomography, we demonstrate that outer hair cells in living guinea pigs have length changes with unexpected timing and magnitudes that depend on the stimulus level in the sensitive cochlea. CONCLUSIONS/SIGNIFICANCE: The level-dependent length change is a necessary condition for directly validating that power is expended by the active process presumed to underlie normal hearing
Hydromechanical Structure of the Cochlea Supports the Backward Traveling Wave in the Cochlea In Vivo
The discovery that an apparent forward-propagating otoacoustic emission (OAE) induced basilar membrane vibration has created a serious debate in the field of cochlear mechanics. The traditional theory predicts that OAE will propagate to the ear canal via a backward traveling wave on the basilar membrane, while the opponent theory proposed that the OAE will reach the ear canal via a compression wave. Although accepted by most people, the basic phenomenon of the backward traveling wave theory has not been experimentally demonstrated. In this study, for the first time, we showed the backward traveling wave by measuring the phase spectra of the basilar membrane vibration at multiple longitudinal locations of the basal turn of the cochlea. A local vibration source with a unique and precise location on the cochlear partition was created to avoid the ambiguity of the vibration source in most previous studies. We also measured the vibration pattern at different places of a mechanical cochlear model. A slow backward traveling wave pattern was demonstrated by the time-domain sequence of the measured data. In addition to the wave propagation study, a transmission line mathematical model was used to interpret why no tonotopicity was observed in the backward traveling wave
Linoleic acid stimulates [Ca2+]i increase in rat pancreatic beta-cells through both membrane receptor- and intracellular metabolite-mediated pathways.
The role of the free fatty acid (FFA) receptor and the intracellular metabolites of linoleic acid (LA) in LA-stimulated increase in cytosolic free calcium concentration ([Ca(2+)]i) was investigated. [Ca(2+)]i was measured using Fura-2 as indicator in rat pancreatic β-cells in primary culture. LA (20 µM for 2 min) stimulated a transient peak increase followed by a minor plateau increase in [Ca(2+)]i. Elongation of LA stimulation up to 10 min induced a strong and long-lasting elevation in [Ca(2+)]i. Activation of FFA receptors by the non-metabolic agonist GW9508 (40 µM for 10 min) resulted in an increase in [Ca(2+)]i similar to that of 2-min LA treatment. Inhibition of acyl-CoA synthetases by Triacsin C suppressed the strong and long-lasting increase in [Ca(2+)]i. The increase in [Ca(2+)]i induced by 2 min LA or GW9508 were fully eliminated by exhaustion of endoplasmic reticulum (ER) Ca(2+) stores or by inhibition of phospholipase C (PLC). Removal of extracellular Ca(2+) did not influence the transient peak increase in [Ca(2+)]i stimulated by 2 min LA or GW9508. The strong and long-lasting increase in [Ca(2+)]i induced by 10 min LA was only partially suppressed by extracellular Ca(2+) removal or thapsigargin pretreatment, whereas remaining elevation in [Ca(2+)]i was eliminated after exhaustion of mitochondrial Ca(2+) using triphenyltin. In conclusion, LA stimulates Ca(2+) release from ER through activation of the FFA receptor coupled to PLC and mobilizes mitochondrial Ca(2+) by intracellular metabolites in β-cells
Norepinephrine protects against cochlear outer hair cell damage and noise-induced hearing loss via α2A-adrenergic receptor
Abstract Background The cochlear sympathetic system plays a key role in auditory function and susceptibility to noise-induced hearing loss (NIHL). The formation of reactive oxygen species (ROS) is a well-documented process in NIHL. In this study, we aimed at investigating the effects of a superior cervical ganglionectomy (SCGx) on NIHL in Sprague-Dawley rats. Methods We explored the effects of unilateral and bilateral Superior Cervical Ganglion (SCG) ablation in the eight-ten weeks old Sprague-Dawley rats of both sexes on NIHL. Auditory function was evaluated by auditory brainstem response (ABR) testing and Distortion product otoacoustic emissions (DPOAEs). Outer hair cells (OHCs) counts and the expression of α2A-adrenergic receptor (AR) in the rat cochlea using immunofluorescence analysis. Cells culture and treatment, CCK-8 assay, Flow cytometry staining and analysis, and western blotting were to explore the mechanisms of SCG fibers may have a protective role in NIHL. Results We found that neither bilateral nor unilateral SCGx protected the cochlea against noise exposure. In HEI-OC1 cells, H2O2-induced oxidative damage and cell death were inhibited by the application of norepinephrine (NE). NE may prevent ROS-induced oxidative stress in OHCs and NIHL through the α2A-AR. Conclusion These results demonstrated that sympathetic innervation mildly affected cochlear susceptibility to acoustic trauma by reducing oxidative damage in OHCs through the α2A-AR. NE may be a potential therapeutic strategy for NIHL prevention
Phenotypic characterization of GPR120-expressing cells in the interstitial tissue of pancreas
GPR120 functions as a plasma membrane receptor for unsaturated long-chain free fatty acids and involves in GLP-1 secretion, adipogenesis and the control of energy balance. Pancreas is the key organ in fuel and energy metabolism. Here GPR120 expression in human and rat pancreas was observed by RT-PCR, and the distribution and phenotypes of GPR120-positive cells in human and rat pancreas were shown by immunohistochemical staining. GPR120 mRNA expression was found in human and rat pancreas. GPR120-positive cells were scattered mainly in the interstitial tissues of human and rat pancreas, and they were not co-localized with nestin, vimentin, alpha-SMA and glucagon, respectively. However, GPR120 was distributed on the cells positively stained by CD68, the specific marker of macrophages, and on the cells positive stained by CD34 and CD117, the markers of interstitial cells. In conclusion, this study demonstrates the expression of GPR120 in pancreas and shows the distribution of GPR120 in human and rat pancreas
Absolute measurement of subnanometer scale vibration of cochlear partition of an excised guinea pig cochlea using spectral-domain phase-sensitive optical coherence tomography
Direct measurement of absolute vibration parameters from different locations within the mammalian organ of Corti is crucial for understanding the hearing mechanics such as how sound propagates through the cochlea and how sound stimulates the vibration of various structures of the cochlea, namely, basilar membrane (BM), recticular lamina, outer hair cells and tectorial membrane (TM). In this study we demonstrate the feasibility a modified phase-sensitive spectral domain optical coherence tomography system to provide subnanometer scale vibration information from multiple angles within the imaging beam. The system has the potential to provide depth resolved absolute vibration measurement of tissue microstructures from each of the delay-encoded vibration images with a noise floor of ∼0.3nm at 200Hz. © 2012 SPIE
In vivo measurement of amplifying motion within the organ of Corti under sound stimulation using optical coherence tomography
Hearing in mammals, depend on an amplifying motion which hypothetically uses force from outer hair cells (OHC) motility to enhance sound induced vibration of the organ of Corti of cochlea. In this hypothesis the differential motion among key structures in this organ and the timing of the OHC force generation is essential for cochlear amplification to occur. Using a time domain optical coherence tomography system which allows us to make vibration measurements we were able to measure differential motion of two functionally important surfaces, namely, basilar membrane and reticular lamina. The reticular lamina vibrates at higher amplitude than the basilar membrane and has significant phase lead over basilar membrane vibration. The differential motion, that is, different amplitude and phase of vibration, become less as the energy of the sound stimulus is increased and the amplification processes in the organ of Corti are quenched. © 2012 SPIE
In vivo measurement of differential motion inside the organ of Corti using a low coherence interferometer system
The differential motion of the organ of Corti has been expected as a result of the outer hair cell force, believed to be necessary for the cochlear amplifier. In vitro experiments have been performed to demonstrate this motion but the in vivo data was unavailable due to the technical difficulties. Using a specially-designed time-domain optical coherence tomography system, we performed in vivo imaging and vibration measurement at the sensitive base of the guinea pig cochlea. This technique, for the first time, provides in vivo information about the internal vibration of the organ of Corti. At low sound level, when the cochlea is more sensitive, top surface of the organ of Corti, the reticular lamina (RL) showed tuning at a higher frequency than of the bottom surface, basilar membrane (BM) and its vibration amplitude is 2-3 times of that of the BM. Corresponding to the frequency difference, the phase of RL vibration is lead to that of the BM. Both the amplitude gain and the phase lead on RL is level dependent. This suggests that they are related to the cochlear amplification. The amplitude gain at the RL is an enhancement of the BM motion for stimulating the stereocillia. The advance in time of RL vibration can prepare proper timing of stereocillia stimulation for the cochlear amplification. © 2012 SPIE
Signal flow inside the tunnel of Corti
All With the advent of Optical Coherence Tomography (OCT), a variation of the standard laser-interferometer technique, vibrations of various points inside the cochlea can be measured separately and concurrently. In this work we measured vibrations of the basilar membrane (BM) and the Reticular Lamina (RL) in the cochlea of the guinea pig. Stimulus tones had frequencies in the range from 10 to 25 kHz, they were generated and measured with a spacing of 250 Hz. By smoothing and interpolation the spacing was reduced to 50 Hz. We confirmed earlier findings in that in viable animals the responses at the RL are generally larger than those of the BM, and have smaller phase delays. Moreover, these differences are little dependent of the level of stimulation. Our main hypothesis is: stimulation of the stapes primarily excites the structures in the upper (RL) part of the Organ of Corti (OoC) channel. Subsequently, movements of the RL cause movements of the fluid in the OoC channel, which in turn moves the BM. Computation of the sound field generated by the RL yielded results that agree very well with the data. These results thus confirm the hypothesis
Effects of inhibiting PLC by U73122 on LA-induced increase in [Ca<sup>2+</sup>]i in rat β-cells.
<p>A: LA-stimulated increase in [Ca<sup>2+</sup>]i in U73122 treatment (10µM for 10min). B: the statistical results of the maximal Ca<sup>2+</sup> levels in the first phase and in the second phase. ** means P<0.01 vs basal level and the first phase. C: The mean Ca<sup>2+</sup> changes in the first phase. ** means P<0.01 vs LA. D: The mean Ca<sup>2+</sup> changes in the second phase (n = 7 for U73122).</p