Multielectrode array use in insect auditory neuroscience to unravel the spatio-temporal response pattern in the prothoracic ganglion of Mecopoda elongata.

Mechanoreceptors in hearing organs transduce sound-induced mechanical responses into neuronal signals, which are further processed and forwarded to the brain along a chain of neurons in the auditory pathway. Bushcrickets (katydids) have their ears in the front leg tibia, and the first synaptic integration of sound-induced neuronal signals takes place in the primary auditory neuropil of the prothoracic ganglion. By combining intracellular recordings of the receptor activity in the ear, extracellular multichannel array recordings on top of the prothoracic ganglion and hook electrode recordings at the neck connective, we mapped the timing of neuronal responses to tonal sound stimuli along the auditory pathway from the ears towards the brain. The use of the multielectrode array allows the observation of spatio-temporal patterns of neuronal responses within the prothoracic ganglion. By eliminating the sensory input from one ear, we investigated the impact of contralateral projecting interneurons in the prothoracic ganglion and added to previous research on the functional importance of contralateral inhibition for binaural processing. Furthermore, our data analysis demonstrates changes in the signal integration processes at the synaptic level indicated by a long-lasting increase in the local field potential amplitude. We hypothesize that this persistent increase of the local field potential amplitude is important for the processing of complex signals, such as the conspecific song.

Variations in cochlea shape reveal different evolutionary adaptations in primates and rodents.

The presence of a coiled cochlea is a unique feature of the therian inner ear. While some aspects of the cochlea are already known to affect hearing capacities, the full extent of the relationships between the morphology and function of this organ are not yet understood-especially when the effect of body size differences between species is minimized. Here, focusing on Euarchontoglires, we explore cochlear morphology of 33 species of therian mammals with a restricted body size range. Using µCT scans, 3D models and 3D geometric morphometrics, we obtained shape information of the cochlea and used it to build phylogenetically corrected least square models with 12 hearing variables obtained from the literature. Our results reveal that different taxonomic groups differ significantly in cochlea shape. We further show that these shape differences are related to differences in hearing capacities between these groups, despite of similar cochlear lengths. Most strikingly, rodents with good low-frequency hearing display "tower-shaped" cochleae, achieved by increasing the degree of coiling of their cochlea. In contrast, primates present relatively wider cochleae and relative better high frequency hearing. These results suggest that primates and rodents increased their cochlea lengths through different morpho-evolutionary trajectories.

Low radiodensity µCT scans to reveal detailed morphology of the termite leg and its subgenual organ.

Termites sense tiny substrate-borne vibrations through subgenual organs (SGOs) located within their legs' tibiae. Little is known about the SGOs' structure and physical properties. We applied high-resolution (voxel size 0.45 µm) micro-computed tomography (µCT) to Australian termites, Coptotermes lacteus and Nasutitermes exitiosus (Hill) to test two staining techniques. We compared the effectiveness of a single stain of Lugol's iodine solution (LS) to LS followed by Phosphotungstic acid (PTA) solutions (1% and 2%). We then present results of a soldier of Nasutitermes exitiosus combining µCT with LS + 2%PTS stains and scanning electron microscopy to exemplify the visualisation of their SGOs. The termite's SGO due to its approximately oval shape was shown to have a maximum diameter of 60 µm and a minimum of 48 µm, covering 60 ± 4% of the leg's cross-section and 90.4 ± 5% of the residual haemolymph channel. Additionally, the leg and residual haemolymph channel cross-sectional area decreased around the SGO by 33% and 73%, respectively. We hypothesise that this change in cross-sectional area amplifies the vibrations for the SGO. Since SGOs are directly connected to the cuticle, their mechanical properties and the geometric details identified here may enable new approaches to determine how termites sense micro-vibrations.

Comparing the electrophysiological effects of traumatic noise exposure between rodents.

Noise-induced hearing deficits are important health problems in the industrialized world. As the underlying physiological dysfunctions are not well understood, research in suitable animal models is urgently needed. Three rodent species (Mongolian gerbil, rat, and mouse) were studied to compare the temporal dynamics of noise-induced hearing loss after identical procedures of noise exposure. Auditory brainstem responses (ABRs) were measured before, during, and up to 8 wk after noise exposure for threshold determination and ABR waveform analysis. Trauma induction with stepwise increasing sound pressure level was interrupted by five interspersed ABR measurements. Comparing short- and long-term dynamics underlying the following noise-induced hearing loss revealed diverging time courses between the three species. Hearing loss occurred early on during noise exposure in all three rodent species at or above trauma frequency. Initial noise level (105 dB SPL) was most effective in rats whereas the delayed level increase to 115 dB SPL affected mice much stronger. Induced temporary threshold shifts in rats and mice were larger in animals with lower pretrauma ABR thresholds. The increase in activity (gain) along the auditory pathway was derived by comparing the amplitudes of short- and long-latency ABR waveform components. Directly after trauma, significant effects were found for rats (decreasing gain) and mice (increasing gain) whereas gerbils revealed high individual variability in gain changes. Taken together, our comparative study revealed pronounced species-specific differences in the development of noise-induced hearing loss and the related processing along the auditory pathway.NEW & NOTEWORTHY We compared deficits after noise trauma in different rodents that are typically used in hearing research (Mongolian gerbil, rat, and mouse). We observed noise-induced threshold changes and alterations in the activity of processing auditory information along the ascending auditory pathway. Our results reveal pronounced differences in the characteristics of trauma-induced damage in these different rodent groups.

Strain Comparison in Rats Differentiates Strain-Specific from More General Correlates of Noise-Induced Hearing Loss and Tinnitus.

Experiments in rodent animal models help to reveal the characteristics and underlying mechanisms of pathologies related to hearing loss such as tinnitus or hyperacusis. However, a reliable understanding is still lacking. Here, four different rat strains (Sprague Dawley, Wistar, Long Evans, and Lister Hooded) underwent comparative analysis of electrophysiological (auditory brainstem responses, ABRs) and behavioral measures after noise trauma induction to differentiate between strain-dependent trauma effects and more consistent changes across strains, such as frequency dependence or systematic temporal changes. Several hearing- and trauma-related characteristics were clearly strain-dependent. Lister Hooded rats had especially high hearing thresholds and were unable to detect a silent gap in continuous background noise but displayed the highest startle amplitudes. After noise exposure, ABR thresholds revealed a strain-dependent pattern of recovery. ABR waveforms varied in detail among rat strains, and the difference was most prominent at later peaks arising approximately 3.7 ms after stimulus onset. However, changes in ABR waveforms after trauma were small compared to consistent strain-dependent differences between individual waveform components. At the behavioral level, startle-based gap-prepulse inhibition (gap-PPI) was used to evaluate the occurrence and characteristics of tinnitus after noise exposure. A loss of gap-PPI was found in 33% of Wistar, 50% of Sprague Dawley, and 75% of Long Evans rats. Across strains, the most consistent characteristic was a frequency-specific pattern of the loss of gap-PPI, with the highest rates at approximately one octave above trauma. An additional range exhibiting loss of gap-PPI directly below trauma frequency was revealed in Sprague Dawley and Long Evans rats. Further research should focus on these frequency ranges when investigating the underlying mechanisms of tinnitus induction.

Tuned vibration modes in a miniature hearing organ: Insights from the bushcricket.

Bushcrickets (katydids) rely on only 20 to 120 sensory units located in their forelegs to sense sound. Situated in tiny hearing organs less than 1 mm long (40× shorter than the human cochlea), they cover a wide frequency range from 1 kHz up to ultrasounds, in tonotopic order. The underlying mechanisms of this miniaturized frequency-place map are unknown. Sensory dendrites in the hearing organ (crista acustica [CA]) are hypothesized to stretch, thereby driving mechanostransduction and frequency tuning. However, this has not been experimentally confirmed. Using optical coherence tomography (OCT) vibrometry, we measured the relative motion of structures within and adjacent to the CA of the bushcricket Mecopoda elongata We found different modes of nanovibration in the CA that have not been previously described. The two tympana and the adjacent septum of the foreleg that enclose the CA were recorded simultaneously, revealing an antiphasic lever motion strikingly reminiscent of vertebrate middle ears. Over the entire length of the CA, we were able to separate and compare vibrations of the top (cap cells) and base (dorsal wall) of the sensory tissue. The tuning of these two structures, only 15 to 60 µm (micrometer) apart, differed systematically in sharpness and best frequency, revealing a tuned periodic deformation of the CA. The relative motion of the two structures, a potential drive of transduction, demonstrated sharper tuning than either of them. The micromechanical complexity indicates that the bushcricket ear invokes multiple degrees of freedom to achieve frequency separation with a limited number of sensory cells.

Comparative analysis of a geometric and an adhesive righting strategy against toppling in inclined hexapedal locomotion.

Animals are known to exhibit different walking behaviors in hilly habitats. For instance, cats, rats, squirrels, tree frogs, desert iguana, stick insects and desert ants were observed to lower their body height when traversing slopes, whereas mound-dwelling iguanas and wood ants tend to maintain constant walking kinematics regardless of the slope. This paper aims to understand and classify these distinct behaviors into two different strategies against toppling for climbing animals by looking into two factors: (i) the torque of the center of gravity (CoG) with respect to the critical tipping axis, and (ii) the torque of the legs, which has the potential to counterbalance the CoG torque. Our comparative locomotion analysis on level locomotion and inclined locomotion exhibited that primarily only one of the proposed two strategies was chosen for each of our sample species, despite the fact that a combined strategy could have reduced the animal's risk of toppling over even more. We found that Cataglyphis desert ants (species Cataglyphis fortis) maintained their upright posture primarily through the adjustment of their CoG torque (geometric strategy), and Formica wood ants (species Formica rufa), controlled their posture primarily by exerting leg torques (adhesive strategy). We further provide hints that the geometric strategy employed by Cataglyphis could increase the risk of slipping on slopes as the leg-impulse substrate angle of Cataglyphis hindlegs was lower than that of Formica hindlegs. In contrast, the adhesion strategy employed by Formica front legs not only decreased the risk of toppling but also explained the steeper leg-impulse substrate angle of Formica hindlegs which should relate to more bending of the tarsal structures and therefore to more microscopic contact points, potentially reducing the risk of hindleg slipping.

Comparative micromechanics of bushcricket ears with and without a specialized auditory fovea region in the crista acustica.

In some insects and vertebrate species, the specific enlargement of sensory cell epithelium facilitates the perception of particular behaviourally relevant signals. The insect auditory fovea in the ear of the bushcricket Ancylecha fenestrata (Tettigoniidae: Phaneropterinae) is an example of such an expansion of sensory epithelium. Bushcricket ears developed in convergent evolution anatomical and functional similarities to mammal ears, such as travelling waves and auditory foveae, to process information by sound. As in vertebrate ears, sound induces a motion of this insect hearing organ (crista acustica), which can be characterized by its amplitude and phase response. However, detailed micromechanics in this bushcricket ear with an auditory fovea are yet unknown. Here, we fill this gap in knowledge for bushcricket, by analysing and comparing the ear micromechanics in Ancylecha fenestrata and a bushcricket species without auditory fovea (Mecopoda elongata, Tettigoniidae: Mecopodinae) using laser-Doppler vibrometry. We found that the increased size of the crista acustica, expanded by a foveal region in A. fenestrata, leads to higher mechanical amplitudes and longer phase delays in A. fenestrata male ears. Furthermore, area under curve analyses of the organ oscillations reveal that more sensory units are activated by the same stimuli in the males of the auditory fovea-possessing species A. fenestrata. The measured increase of phase delay in the region of the auditory fovea supports the conclusion that tilting of the transduction site is important for the effective opening of the involved transduction channels. Our detailed analysis of sound-induced micromechanics in this bushcricket ear demonstrates that an increase of sensory epithelium with foveal characteristics can enhance signal detection and may also improve the neuronal encoding.

Experimental and Theoretical Explorations of Traveling Waves and Tuning in the Bushcricket Ear.

The ability to detect airborne sound is essential for many animals. Examples from the inner ear of mammals and bushcrickets demonstrate that similar detection strategies evolved in taxonomically distant species. Both mammalian and bushcricket ears possess a narrow strip of sensory tissue that exhibits an anatomical gradient and traveling wave motion responses used for frequency discrimination. We measured pressure and motion in the bushcricket ear to investigate physical properties, stiffness, and mass, which govern the mechanical responses to sound. As in the mammalian cochlea, sound-induced fluid pressure and motion responses were tonotopically organized along the longitudinal axis of the crista acustica, the bushcricket's hearing organ. The fluid pressure at the crista and crista motion were used to calculate the acoustic impedance of the organ-bounded fluid mass (Zmass). We used a theoretical wave analysis of wavelength data from a previous study to predict the crista acustica stiffness. The wave analysis also predicts Zmass, and that result agreed reasonably well with the directly measured Zmass, lending support to the theoretical wave analysis. The magnitude of the crista stiffness was similar to basilar membrane stiffness in mammals, and as in mammals, the stiffness decreased from the high-frequency to the low-frequency region. At a given location, the stiffness increased with increasing frequency, corresponding to increasing curvature of the traveling wave (decreasing wavelength), indicating that longitudinal coupling plays a substantial role in determining crista stiffness. This is in contrast to the mammalian ear, in which stiffness is independent of frequency and longitudinal coupling is relatively small.

Functional basis of the sexual dimorphism in the auditory fovea of the duetting bushcricket Ancylecha fenestrata.

From mammals to insects, acoustic communication is in many species crucial for successful reproduction. In the duetting bushcricket Ancylecha fenestrata, the mutual acoustic communication between males and females is asymmetrical. We investigated how those signalling disparities are reflected by sexual dimorphism of their ears. Both sexes have tympanic ears in their forelegs, but male ears possess a significantly longer crista acustica containing 35% more scolopidia. With more sensory cells to cover a similar hearing range, the male hearing organ shows a significantly expanded auditory fovea that is tuned to the dominant frequency of the female reply to facilitate phonotactic mate finding. This sex-specific auditory fovea is demonstrated in the mechanical and neuronal responses along the tonotopically organized crista acustica by laservibrometric and electrophysiological frequency mapping, respectively. Morphometric analysis of the crista acustica revealed an interrupted gradient in organ height solely within this auditory fovea region, whereas all other anatomical parameters decrease continuously from proximal to distal. Combining behavioural, anatomical, biomechanical and neurophysiological information, we demonstrate evidence of a pronounced auditory fovea as a sex-specific adaptation of an insect hearing organ for intraspecific acoustic communication.

Hearing Without Neuroglobin.

Neuroglobin (Ngb) is a member of the globin family of respiratory proteins, which was recently observed in many neurons of the auditory pathways. Up to now, however, nothing was known about the role of Ngb in hearing processes. We therefore studied auditory function by recording distortion-product otoacoustic emissions (DPOAE) and auditory brainstem responses (ABRs) in wild-type (C57BL/6N) and Ngb-knockout mice. In KO mice, DPOAE thresholds were moderately augmented in the range of 5-18 kHz, reaching statistical significance at 8 and 10 kHz, while the ABR thresholds were not different between groups. The activation of the efferent system by an additional noise given to the contralateral ear resulted in an increased f2-f1-emission level only in WT animals. A noise exposure resulted in similar acute threshold shifts in the DPOAE and ABR of both animal groups. The recovery of hearing function, expressed by decreased DPOAE thresholds, was not significantly different between groups after four days and after four weeks. ABR recordings showed that threshold shifts elicited by noise-trauma were slightly better revised in wild-type mice. While ABR amplitudes were similar in both groups before noise overexposure, four weeks after trauma a moderate but statistically significant decrease of the latest peak-to-peak response amplitude (originating in the inferior colliculus) was observed in KO mice. Our results suggest that the lack of Ngb, at least in the model used in the present study, results in only marginal deficits in hearing ability. A putative functional role of Ngb in the efferent system warrants further studies.

Morphological basis for a tonotopic design of an insect ear.

The tonotopically organized hearing organs of bushcrickets provide the opportunity for a detailed correlation of morphological and structural properties within hearing organs that are needed to establish tonotopic gradients. In the present study of a tonotopic insect hearing organ, we combine mechanical measurements of sound-induced hearing organ motion and detailed anatomical investigations to explore the anatomical basis of tonotopy. We compare mechanical data of frequency responses along the auditory organ to several anatomical parameters. Low frequency responses are related to larger organ and cap cell size in the proximal part of the hearing organ while in the distal part of the organ, small organ and cap cell size is related to high-frequency representation. However, the correlation between organ and cap cell size with continuous frequency representation along the organ is not very tight. Instead, the height of the organ and the corresponding length of the sensory dendrites are best correlated to tonotopic frequency representation. The sensory dendrite contains a ciliary root with a pronounced cross-banding of electron-dense material that should be important for the stiffness of the dendrite. The geometry of surrounding structures like the hemolymph channel and the acoustic trachea as well as the extension of the tectorial membrane are not correlated to the tonotopy. We provide evidence that tonotopy in the bushcricket hearing organ may depend on the size of ciliary structures. In particular, the ciliary root of the sensory cells is a likely cellular basis of tonotopy.

Auditory fovea in the ear of a duetting katydid shows male-specific adaptation to the female call.

Convergent evolution has led to surprising functional and mechanistic similarities between the vertebrate cochlea and some katydid ears [1,2]. Here we report on an 'auditory fovea' (Figure 1A) in the duetting katydid Ancylecha fenestrata (Tettigoniidae). The auditory fovea is a specialized inner-ear region with a disproportionate number of receptor cells tuned to a narrow frequency range, and has been described in the cochlea of some vertebrates, such as bats and mole rats [3,4]. In tonotopically organized ears, the location in the hearing organ of the optimal neuronal response to a tone changes gradually with the frequency of the stimulation tone. However, in the ears of A. fenestrata, the sensory cells in the auditory fovea are tuned to the dominant frequency of the female call; this area of the hearing organ is extensively expanded in males to provide an overrepresentation of this behaviorally important auditory input. Vertebrates developed an auditory fovea for improved prey or predator detection. In A. fenestrata, however, the foveal region facilitates acoustic pair finding, and the sexual dimorphism of sound-producing and hearing organs reflects the asymmetry in the mutual communication system between the sexes (Figures 1B, S1).

Dependence of the Startle Response on Temporal and Spectral Characteristics of Acoustic Modulatory Influences in Rats and Gerbils.

The acoustic startle response (ASR) and its modulation by non-startling prepulses, presented shortly before the startle-eliciting stimulus, is a broadly applied test paradigm to determine changes in neural processing related to auditory or psychiatric disorders. Modulation by a gap in background noise as a prepulse is especially used for tinnitus assessment. However, the timing and frequency-related aspects of prepulses are not fully understood. The present study aims to investigate temporal and spectral characteristics of acoustic stimuli that modulate the ASR in rats and gerbils. For noise-burst prepulses, inhibition was frequency-independent in gerbils in the test range between 4 and 18 kHz. Prepulse inhibition (PPI) by noise-bursts in rats was constant in a comparable range (8-22 kHz), but lower outside this range. Purely temporal aspects of prepulse-startle-interactions were investigated for gap-prepulses focusing mainly on gap duration. While very short gaps had no (rats) or slightly facilitatory (gerbils) influence on the ASR, longer gaps always had a strong inhibitory effect. Inhibition increased with durations up to 75 ms and remained at a high level of inhibition for durations up to 1000 ms for both, rats and gerbils. Determining spectral influences on gap-prepulse inhibition (gap-PPI) revealed that gerbils were unaffected in the limited frequency range tested (4-18 kHz). The more detailed analysis in rats revealed a variety of frequency-dependent effects. Gaps in pure-tone background elicited constant and high inhibition (around 75%) over a broad frequency range (4-32 kHz). For gaps in noise-bands, on the other hand, a clear frequency-dependency was found: inhibition was around 50% at lower frequencies (6-14 kHz) and around 70% at high frequencies (16-20 kHz). This pattern of frequency-dependency in rats was specifically resulting from the inhibitory effect by the gaps, as revealed by detailed analysis of the underlying startle amplitudes. An interaction of temporal and spectral influences, finally, resulted in higher inhibition for 500 ms gaps than for 75 ms gaps at all frequencies tested. Improved prepulse paradigms based on these results are well suited to quantify the consequences of central processing disorders.

Gating of Acoustic Transducer Channels Is Shaped by Biomechanical Filter Processes.

Mechanoelectrical transduction of acoustic signals is the fundamental process for hearing in all ears across the animal kingdom. Here, we performed in vivo laser-vibrometric and electrophysiological measurements at the transduction site in an insect ear (Mecopoda elongata) to relate the biomechanical tonotopy along the hearing organ to the frequency tuning of the corresponding sensory cells. Our mechanical and electrophysiological map revealed a biomechanical filter process that considerably sharpens the neuronal response. We demonstrate that the channel gating, which acts on chordotonal stretch receptor neurons, is based on a mechanical directionality of the sound-induced motion. Further, anatomical studies of the transduction site support our finding of a stimulus-relevant tilt. In conclusion, we were able to show, in an insect ear, that directionality of channel gating considerably sharpens the neuronal frequency selectivity at the peripheral level and have identified a mechanism that enhances frequency discrimination in tonotopically organized ears.

Processing of simple and complex acoustic signals in a tonotopically organized ear.

Processing of complex signals in the hearing organ remains poorly understood. This paper aims to contribute to this topic by presenting investigations on the mechanical and neuronal response of the hearing organ of the tropical bushcricket species Mecopoda elongata to simple pure tone signals as well as to the conspecific song as a complex acoustic signal. The high-frequency hearing organ of bushcrickets, the crista acustica (CA), is tonotopically tuned to frequencies between about 4 and 70 kHz. Laser Doppler vibrometer measurements revealed a strong and dominant low-frequency-induced motion of the CA when stimulated with either pure tone or complex stimuli. Consequently, the high-frequency distal area of the CA is more strongly deflected by low-frequency-induced waves than by high-frequency-induced waves. This low-frequency dominance will have strong effects on the processing of complex signals. Therefore, we additionally studied the neuronal response of the CA to native and frequency-manipulated chirps. Again, we found a dominant influence of low-frequency components within the conspecific song, indicating that the mechanical vibration pattern highly determines the neuronal response of the sensory cells. Thus, we conclude that the encoding of communication signals is modulated by ear mechanics.

Mechanical basis of otoacoustic emissions in tympanal hearing organs.

Tympanal hearing organs of insects emit distortion-product otoacoustic emissions (DPOAEs), which in mammals are used as indicator for nonlinear cochlear amplification, and which are highly vulnerable to manipulations interfering with the animal's physiological state. Although in previous studies, evidence was provided for the involvement of auditory mechanoreceptors, the source of DPOAE generation and possible active mechanisms in tympanal organs remained unknown. Using laser Doppler vibrometry in the locust ear, we show that DPOAEs mechanically emerge at the tympanum region where the auditory mechanoreceptors are attached. Those emission-coupled vibrations differed remarkably from tympanum waves evoked by external pure tones of the same frequency, in terms of wave propagation, energy distribution, and location of amplitude maxima. Selective inactivation of the auditory receptor cells by mechanical lesions did not affect the tympanum's response to external pure tones, but abolished the emission's displacement amplitude peak. These findings provide evidence that tympanal auditory receptors, comparable to the situation in mammals, comprise the required nonlinear response characteristics, which during two-tone stimulation lead to additional, highly localized deflections of the tympanum.

Lateralization of travelling wave response in the hearing organ of bushcrickets.

Travelling waves are the physical basis of frequency discrimination in many vertebrate and invertebrate taxa, including mammals, birds, and some insects. In bushcrickets (Tettigoniidae), the crista acustica is the hearing organ that has been shown to use sound-induced travelling waves. Up to now, data on mechanical characteristics of sound-induced travelling waves were only available along the longitudinal (proximal-distal) direction. In this study, we use laser Doppler vibrometry to investigate in-vivo radial (anterior-posterior) features of travelling waves in the tropical bushcricket Mecopoda elongata. Our results demonstrate that the maximum of sound-induced travelling wave amplitude response is always shifted towards the anterior part of the crista acustica. This lateralization of the travelling wave response induces a tilt in the motion of the crista acustica, which presumably optimizes sensory transduction by exerting a shear motion on the sensory cilia in this hearing organ.

Mechanical tuning of the moth ear: distortion-product otoacoustic emissions and tympanal vibrations.

The mechanical tuning of the ear in the moth Empyreuma pugione was investigated by distortion-product otoacoustic emissions (DPOAE) and laser Doppler vibrometry (LDV). DPOAE audiograms were assessed using a novel protocol that may be advantageous for non-invasive auditory studies in insects. To evoke DPOAE, two-tone stimuli within frequency and level ranges that generated a large matrix of values (960 frequency-level combinations) were used to examine the acoustic space in which the moth tympanum shows its best mechanical and acoustical responses. The DPOAE tuning curve derived from the response matrix resembles that obtained previously by electrophysiology, and is V-shaped and tuned to frequencies between 25 and 45 kHz with low Q10dB values of 1.21±0.26. In addition, while using a comparable stimulation regime, mechanical distortion in the displacement of the moth's tympanal membrane at the stigma was recorded with a laser Doppler vibrometer. The corresponding mechanical vibration audiograms were compared with DPOAE audiograms. Both types of audiograms have comparable shape, but most of the mechanical response fields are shifted towards lower frequencies. We showed for the first time in moths that DPOAE have a pronounced analogy in the vibration of the tympanic membrane where they may originate. Our work supports previous studies that point to the stigma (and the internally associated transduction machinery) as an important place of sound amplification in the moth ear, but also suggests a complex mechanical role for the rest of the transparent zone.

Temperature dependence of distortion-product otoacoustic emissions in tympanal organs of locusts.

Distortion-product otoacoustic emissions (DPOAEs) in tympanal organs of insects are vulnerable to manipulations that interfere with the animal's physiological state. Starting at a medium temperature, we raised and lowered the locust's body temperature within the range of 12 to 35°C by changing the temperature of the surrounding air, while recording DPOAEs. These experimental manipulations resulted in reversible amplitude changes of the 2f(1)-f(2) emission, which were dependent on stimulus frequency and level. Using low f(2) frequencies of up to 10 kHz, a temperature increase (median +8-9°C) led to an upward shift of DPOAE amplitudes of approximately +10 dB, whereas a temperature decrease (median -7°C) was followed by a reduction of DPOAE amplitudes by 3 to 5 dB. Both effects were only present in the range of the low-level component of DPOAE growth functions below L2 levels (levels of the f(2) stimulus) of approximately 30 dB SPL. DPOAEs evoked by higher stimulus levels as well as measurements using higher stimulation frequencies above 10 kHz remained unaffected by any temperature shifts. The Arrhenius activation energy was calculated from the -10 dB SPL thresholds (representing the low-level component) of growth functions, which had been measured with 8 and 10 kHz as f(2) frequencies and amounted to up to ~34 and 41 kJ mol(-1), respectively. Such activation energy values provide a hint that the dynein-tubulin system within the scolopidial receptors could play an essential part in the DPOAE generation in tympanal organs.