Scala Vestibuli Research Articles

Longitudinal endolymph movements induced by perilymphatic injections

Endolymph movements and endocochlear potential (EP) changes were measured during disturbances of perilymphatic pressure, induced by injecting artificial perilymph into scala tympani (ST) or scala vestibuli (SV) of the guinea pig cochlea. Injections were performed either with or without an outlet made in the opposite perilymphatic scala. Injections into ST without an outlet induced large pressure changes but virtually no endolymph movement or EP change. Injection at the same rate into ST with an outlet in SV produced smaller pressure changes which were accompanied by a basally-directed displacement of endolymph and significant EP changes. The magnitude of endolymph displacements and EP changes varied as a function of injection rate. Injections into SV, either with or without an outlet in ST, produced apically-directed endolymph displacement and EP changes. For the SV injections without an outlet, the cochlear aqueduct and round window are likely to provide an outlet and compliance, permitting flow along the perilymphatic scalae to occur even when no ST outlet was provided. We conclude that endolymph movements are not dependent on the absolute pressure of the perilymph, but instead occur when small, sustained pressure gradients are present across the cochlear partition, corresponding to times when perilymph flow is induced. This study demonstrates that in the normal, sealed cochlea, endolymph and EP are insensitive to fluid injections into ST, but are sensitive to fluid injections into SV. Endolymph movements are therefore unlikely to be generated by cerebrospinal fluid pressure fluctuations (such as those produced by respiration, posture changes, coughing, sneezing, etc) which are transmitted to ST by the cochlear aqueduct.

Enhancement of electrically evoked oto-acoustic emissions associated with low-frequency stimulus bias of the basilar membrane towards scala vestibuli.

Electrically evoked oto-acoustic emissions (EEOAEs) are sounds present in the ear canal when ac current is passed into the cochlea. EEOAEs are attributed to the activation of fast electromotile responses in outer hair cells (OHCs). An interesting property of EEOAEs is the phenomenon of "acoustic enhancement," where the emission amplitude is increased by moderate-level sound [D. C. Mountain and A. E. Hubbard, Hear. Res. 42, 195-202 (1989)]. In this report a form of enhancement is described which occurs with displacements of the basilar membrane toward scala vestibuli, during amplitude modulation of the EEOAE waveform by low-frequency tones. This "SV-bias enhancement" possibly consists of two components: (i) a low-level component induced by sound at levels which produce nonlinear growth of the cochlear microphonic and which may be equivalent to the "acoustic enhancement" described previously, and (ii) a high-level component which occurs at sound levels well above those which cause saturation of the cochlear microphonic. The low-level component could be explained by either an increased access of the extrinsically applied current to a membrane-based source of OHC motility, perhaps coupled with a reduction in negative feedback, or an increase in electromotile output during scala vestibuli displacements, but the origin of the high-level component is obscure.

Changes in the subcellular localization of the Brn4 gene product precede mesenchymal remodeling of the otic capsule

To better understand the genetic mechanisms that regulate the formation of the temporal bone, we have characterized the developmental expression pattern of the mouse gene, Brn4/Pou3f4, which plays a central role in bony labyrinth formation. Expression of this gene is initially detected in the ventral aspect of the otic capsule at 10.5 days post coitus (dpc), and correlates with the onset of mesenchymal condensation in the otic capsule. As the otic capsule condenses further and surrounds the entire otic vesicle, the Brn4 gene product is detected throughout the inner ear in the mesenchyme of both the cochlear and vestibular aspects. Early in otic embryogenesis, the Brn4 gene product is localized to the nucleus of the vast majority of cells in which it is expressed. The Brn4 gene product remains nuclear in those regions of the otic capsule that eventually give rise to the mature bony labyrinth. However, the subcellular localization of the Brn4 gene product shifts from strictly nuclear to perinuclear in those regions of the otic capsule that will cavitate to form acellular regions in the temporal bone, such as the scala tympani, scala vestibuli, and the internal auditory meatus. These data provide a detailed analysis of the expression pattern of the Brn4 gene, and provide insight into the role of the Brn4 gene product and its regulation during otic capsule formation.

Localization of mRNA encoding the P2X2 receptor subunit of the adenosine 5?-triphosphate-gated ion channel in the adult and developing rat inner ear by in situ hybridization

Localization of expression of the adenosine 5'-triphosphate (ATP)-gated ion channel P2X2 receptor subunit (P2X2R) in the rat inner ear at different stages of development was achieved by using in situ mRNA hybridization. In the adult, P2X2R mRNA was strongly expressed in many of the cells bordering the cochlear endolymphatic compartment. This included the interdental cells of the spiral limbus, all cells of the inner sulcus and organ of Corti, and cells of the spiral prominence. In the vestibular labyrinth, strong expression was noted in the transitional cells at the base of the crista ampullaris and in the sensory epithelium of the crista and maculae. During development, P2X2R mRNA expression was evident in the precursors of these structures at the earliest period studied, embryonic day 12 (E12). Expression increased during the ontogeny in both the cochlear and the vestibular end organs. In addition, both the spiral and vestibular ganglia showed developmental expression. In contrast to the supporting cells of the organ of Corti, both inner and outer hair cells exhibited P2X2R mRNA only after postnatal day 10 (P10) through P12, concomitant with the onset of hearing. P2X2R expression levels in all cells fell from a maximum at P12-P18 to lower levels in the adult. In the adult, P2X2R mRNA levels were modest in outer hair cells in the basal (high-frequency) encoding region of the cochlea, and inner hair cell labeling was low throughout the cochlea. Reissner's membrane, which maintains an electrochemical barrier between scala vestibuli and scala media, showed considerable expression of P2X2R mRNA in early postnatal development, and expression was maintained at moderate levels in the adult cochlea. These data are consistent with a role for the P2X2R subunit in the processes of labyrinthine development and the regulation of the electrochemical gradients supporting auditory and vestibular sensory transduction.

Localization of mRNA encoding the P2X2 receptor subunit of the adenosine 5′‐triphosphate‐gated ion channel in the adult and developing rat inner ear by in situ hybridization

Localization of expression of the adenosine 5′-triphosphate (ATP)-gated ion channel P2X2 receptor subunit (P2X2R) in the rat inner ear at different stages of development was achieved by using in situ mRNA hybridization. In the adult, P2X2R mRNA was strongly expressed in many of the cells bordering the cochlear endolymphatic compartment. This included the interdental cells of the spiral limbus, all cells of the inner sulcus and organ of Corti, and cells of the spiral prominence. In the vestibular labyrinth, strong expression was noted in the transitional cells at the base of the crista ampullaris and in the sensory epithelium of the crista and maculae. During development, P2X2R mRNA expression was evident in the precursors of these structures at the earliest period studied, embryonic day 12 (E12). Expression increased during the ontogeny in both the cochlear and the vestibular end organs. In addition, both the spiral and vestibular ganglia showed developmental expression. In contrast to the supporting cells of the organ of Corti, both inner and outer hair cells exhibited P2X2R mRNA only after postnatal day 10 (P10) through P12, concomitant with the onset of hearing. P2X2R expression levels in all cells fell from a maximum at P12–P18 to lower levels in the adult. In the adult, P2X2R mRNA levels were modest in outer hair cells in the basal (high-frequency) encoding region of the cochlea, and inner hair cell labeling was low throughout the cochlea. Reissner's membrane, which maintains an electrochemical barrier between scala vestibuli and scala media, showed considerable expression of P2X2R mRNA in early postnatal development, and expression was maintained at moderate levels in the adult cochlea. These data are consistent with a role for the P2X2R subunit in the processes of labyrinthine development and the regulation of the electrochemical gradients supporting auditory and vestibular sensory transduction. J. Comp. Neurol. 393:403–414, 1998. © 1998 Wiley-Liss, Inc.

Possible Ca 2+ -dependent mechanism of apical outer hair cell modulation within the cochlea of the guinea pig

Calcium ions were precipitated with potassium antimonate after injection of the inorganic calcium channel blocker MnCl2 or the inorganic potassium channel blockers BaCl2 or CsCl into the perilymph of the scala vestibuli of the guinea pig. The spatial distribution of the formed histochemical reaction products within the organ of Corti was studied by energy-filtering transmission-electron microscopy. Compared with untreated control ears, the number of the formed precipitates drastically increased at the extracellular side of the lamina reticularis after application of the various inorganic channel blockers. The apical side of the outer hair cells and the intervening Deiter cells were covered by a thick layer of calcium precipitates, whereas the number of histochemical reaction products was clearly reduced in the nearby acellular tectorial membrane. The high calcium content within the formed reaction products at the lamina reticularis could be demonstrated by elemental mapping and by electron energy-loss spectroscopy. To ascertain the alterations in the amounts of the calcium precipitates within the tectorial membrane after application of the various inorganic channel blockers, the precipitate densities were determined semiquantitatively by an image processing system and the values obtained compared with those of untreated control specimens. The observed histochemical results are in good agreement with published electrophysiological findings concerning the spatial distribution of ion channels located at the apical outer hair cell membrane. The detected alterations in the spatial distribution of calcium precipitates might correspond to calcium-dependent processes involved in outer hair cell modulation.

Cochlear Electrically Evoked Emissions Modulated by Mechanical Transduction Channels

Cochlear outer hair cells are capable of both mechanical-to-electrical and electrical-to-mechanical transduction. Vibration of their stereocilia by sound is believed to stimulate somatic motility via a receptor potential developed across the basolateral membrane, thereby enhancing the mechanical vibration and increasing the sensitivity and frequency selectivity of the ear. Extrinsic electrical currents, applied at the tops of the cells, also appear to activate motility in vivo, presumably after entering the cell. Earlier experiments suggested such currents might enter through the transduction channels themselves, but an alternative shunt pathway through the membrane capacitance seems more likely on physical grounds. We therefore recorded electrically evoked oto-acoustic emissions while modulating the transduction channels by driving them with low-frequency sound. Recordings of the low-frequency cochlear microphonic provided a measure of the mean electrical conductance through the channels during sound stimulation. Emissions increased during displacement of the basilar membrane toward scala vestibuli, when the channels were biased open, and decreased on the opposite phase, and the modulation of the emission was in direct proportion to the cochlear microphonic. The results are the strongest evidence yet that electrically evoked emissions are generated directly by mechanisms related to cochlear transduction and lead to the surprising conclusion that, for frequencies up to at least 12 kHz, extrinsic electrical currents enter the hair cell predominantly by the resistive pathway through the transduction channels. Alternatively, the results might be consistent with direct modulation of a motility source driven by capacitive currents but whose output depends on the state of the channels.

一過性虚血後の聴力障害と内耳グルタミン酸

Cochlear compound action potentials (CAPS) and glutamate efflux in the perilymph before, during and after transient cochlear ischemia were studied in the gerbil to reveal the mechanisms of ischemia-induced inner ear damage. Cochlear ischemia was created by occluding the bilateral vertebral arteries for 5 minutes, since the posterior cerebral communicating arteries are lacking and the labyrinthine arteries are solely nourished by the vertebral arteries in the gerbil. Occlusion of the vertebral arteries caused a remarkable increase in the CAP threshold, which subsequently recovered following release of occlusion in all 10 animals. The threshold for acoustic stimulation at 60 minutes after recirculation was, however, higher than that in the preischemic state. Glutamate concentration in the perilymph was measured using a microdialysis technique followed by enzyme cycling analysis. Before occlusion, the glutamate level of the perilymph was 0.35±0.22 pmol/μl in the scala tympani (n=12) and 0.42±0.32 pmol/μl in the scala vestibuli (n=6). Ischemic insult caused an immediate rise in the glutamate level, which rose further even after recirculation, reaching a maximum of 11.72 pmol/μl in the scala tympani (n=12) and 8.49 pmol/μl in the scala vestibuli (n=6). In four animals, the cochlea was processed for transmission electron microscopic study at 30 minutes after ischemic load. The ischemia induced swelling of the afferent auditory nerve endings and vacuolation of the inner hair cells. These results suggest that the release of glutamate in the perilymph may aggravate ischemia-induced hearing dysfunction.

Failure of Forskolin to Elevate the Endocochlear Potential in Experimental Endolymphatic Hydrops of the Guinea Pig

The effect of forskolin (FSK) on the endocochlear potential (EP) in scala media (SM) was examined in experimental endolymphatic hydrops of the guinea pig. Two weeks after obliteration of the endolymphatic sac the EP of hydroptic ears and that of the contralateral control ears were measured by means of microelectrodes. The perfusion of scala vestibuli (SV) with FSK (200 w M) produced EP elevation in the contralateral control ears but failed to do so in the experimental hydroptic ears. Histological examination of experimental endolymphatic hydrops showed mild hydrops with intact appearance of outer and inner hair cells, and the stria vascularis. The mechanism underlying the failure of FSK to elevate the EP in experimental endolymphatic hydrops is discussed.

The Effect of Forskolin on the Sound-evoked Potentials in the Guinea Pig Cochlea

The effect of forskolin (FSK) on cochlear sound-evoked potentials was examined in the guinea pig. The perfusion of the scala vestibuli (SV) with FSK (2 2 10 -4 M) produced a significant increase in the amplitude of negative summating potential ( m SP) with no change in cochlear microphonics (CM) amplitude, and a significant decrease in the amplitude of compound action potential (CAP) with a significant prolongation of N1 latency and a 20 dB CAP threshold elevation. The results lead us to speculate that FSK-induced changes may be involved in the transient formation of endolymphatic hydrops.

Low-frequency suppression of auditory nerve responses to characteristic frequency tones

The effects of low-frequency (50, 100, 200 and 400 Hz) ‘suppressor’ tones on responses to moderate-level characteristic frequency (CF) tones were measured in chinchilla auditory nerve fibers. Two-tone interactions were evident at suppressor intensities of 70–100 dB SPL. In this range, the average response rate decreased as a function of increasing suppressor level and the instantaneous response rate was modulated periodically. At suppression threshold, the phase of suppression typically coincided with basilar membrane displacement toward scala tympani, regardless of CF. At higher suppressor levels, two suppression maxima coexisted, synchronous with peak basilar membrane displacement toward scala tympani and scala vestibuli. Modulation and rate-suppression thresholds did not vary as a function of spontaneous activity and were only minimally correlated with fiber sensitivity. Except for fibers with CF<1 kHz, modulation and rate-suppression thresholds were lower than rate and phase-locking thresholds for the suppressor tones presented alone. In the case of high-CF fibers with low spontaneous activity, excitation thresholds could exceed suppression thresholds by more than 30 dB. The strength of modulation decreased systematically with increasing suppressor frequency. For a given suppressor frequency, modulation was strongest in high-CF fibers and weakest in low-CF fibers. The present findings strongly support the notion that low-frequency suppression in auditory nerve fibers largely reflects an underlying basilar membrane phenomenon closely related to compressive non-linearity.

Pressure-induced basilar membrane position shifts and the stimulus-evoked potentials in the low-frequency region of the guinea pig cochlea.

We have used the guinea pig isolated temporal bone preparation to investigate changes in the non-linear properties of the tone-evoked cochlear potentials during reversible step displacements of the basilar membrane towards either the scala tympani or the scala vestibuli. The position shifts were produced by changing the hydrostatic pressure in the scala tympani. The pressures involved were calculated from measurements of the fluid flow through the system, and the cochlear DC impedance calculated (1.5 x 10(11) kg m-4 s-1, n = 10). Confocal microscopic visualization of the organ of Corti showed that pressure increases in the scala tympani caused alterations of the position of the reticular lamina and stereocilia bundles. For low pressures, there was a sigmoidal relation between the DC pressure applied to the scala tympani (and thus the position shift of the organ of Corti) and the amplitude of the summating potential. The cochlear microphonic potential also showed a pronounced dependence on the applied pressure: pressure changes altered the amplitude of the fundamental as well as its harmonics. In addition, the sound pressure level at which the responses began to saturate was increased, implying a transition towards a linear behaviour. An increase of the phase lag of the cochlear microphonic potential was seen when the basilar membrane was shifted towards the scala vestibuli. We have also measured the intracochlear DC pressure using piezoresistive pressure transducers. The results are discussed in terms of changes in the non-linear properties of cochlear transduction. In addition, the implications of these results for the pathophysiology and diagnosis of Meniérè's disease are discussed.

Reverse middle-ear transfer function in the guinea pig measured with cubic difference tones

Otoacoustic emissions are increasingly useful for determining cochlear function noninvasively. It is widely agreed that these acoustic signals reflect micromechanical processes in the cochlea. However, their quantitative interpretation requires knowledge of the ways in which vibrations travelling back to the ear canal from the cochlea are shaped by the middle ear. An intracochlear source is needed to derive the reverse middle-ear transfer function (rMETF) by comparing pressure in the external ear canal to the corresponding pressure in scala vestibuli. In the present study, the rMETF was obtained in vivo in the guinea pig using as intracochlear sound source the cubic difference tones (CDTs) generated by a pair of external pure tones. With a closed ear canal and open bulla, the rMETF was found to be flat (−35 dB) over a broad frequency range (1.5–8 kHz). The differences between forward and reverse METF could be explained by different loads acting on the middle ear network, which depends on the direction of signal transmission. With knowledge of the rMETF, it becomes possible to quantify CDTs within the cochlea by measuring them noninvasively in the ear canal.

Micro-magnetic resonance imaging of the inner ear in the guinea pig.

We applied a magnetic resonance microscopy at 7.05 T with a gradient coil unit to image the fine structure of the guinea pig cochlea. First, a three-dimensional MR image of the surface of the cochlea was obtained to select the location of cross-sectional images. Then, cross-sectional images of the basal turn, the second turn and the apical turn of the cochlea were obtained. Based upon the different protein concentrations of the endolymph vs the perilymph, the scala vestibuli, scala tympani and the cochlear duct could be clearly distinguished. This allowed a determination of the location of both the basilar membrane and Reissner's membrane. We raise the possibility that MRM may become useful in the diagnosis of endolymphatic hydrops (Meniere's disease).

4-Aminopyridine effects on summating potentials in the guinea pig

DC receptor potentials measured in hair cells, and the associated extracellular DC potential known as the summating potential (SP), originate with nonlinear elements in the mechanoelectric transduction chain. Nonlinear electric conductance has been demonstrated in the basolateral membrane of the hair cell, and is commonly attributed to the presence of voltage- and time-dependent K + conductances in this part of the hair cell membrane. To study a possible contribution of these K + channels to the SP we perfused the perilymphatic spaces of the guinea pig cochlea with the K + channel blocker 4-aminopyridine (4-AP). Since 4-AP might also affect the afferent fibers and, thus, interfere with SP measurement, we added tetrodotoxin (TTX) to the perfusion solutions to block the neuronal discharges. Sound-evoked (2–12 kHz) intracochlear potentials were recorded from the basal turn of both scala vestibuli and scala tympani. The results showed a frequency- and level-dependent effect of 4-AP on the magnitude of the SP. At low and moderate levels of 8 and 12 kHz stimuli 4-AP mostly reduced the SP amplitude, while at high levels of these stimuli and at all levels of 2 and 4 kHz stimuli 4-AP enlarged the SP amplitude. These effects were reversible and occurred in both scala vestibuli and scala tympani. We attribute these bi-directional effects on the SP amplitude to a differential effect of 4-AP on inner hair cell (IHC) and outer hair cell (OHC) physiology. The decrease in SP was found for stimulus conditions where the SP presumably depends mainly on contributions from basal turn IHCs. Blocking the 4-AP-sensitive K + channel in the IHC membrane should lead to a reduced contribution from the IHCs to the SP, because of an increase in basolateral membrane resistance. The increase in SP was found for stimulus conditions where the SP is assumed to depend mainly on contributions from basal turn OHCs. In this case the OHCs seemed to respond to blocking of the 4-AP-sensitive K + channel in the basolateral membrane with an increased contribution to the nonlinearity of the transduction chain. Administration of 4-AP did not affect the endocochlear potential. Light microscopic examination revealed no apparent changes in morphology after 4-AP perfusion.

A new immunohistochemical method for the detection of gentamicin in inner ear fluid compartments.

A new method was developed for frozen section detection of antigens that natively occur in the cochlear peri- and endolymph. A combination of immuno-histochemistry and immunoblot assay enabled topological and quantitative detection of small and hydrophilic molecules (such as the aminoglycoside antibiotics) in frozen sections of the inner ear compartments (scala tympani, scala vestibuli and cochlear duct). A selective localization is possible in the peri- and endolymphatic region of each coil of the cochlea. During sectioning of the cochlea, a small piece of a nitrocellulose membrane is placed to the surface of the intersection and briefly warmed. The sections are cut, simultaneously attached to a nitrocellulose membrane on which the aminoglycoside antibiotics remain adsorbed without any fixation procedure. Using this method, immunoincubation to detect gentamicin was performed in a way usually done in western blot analysis. Results with two different enzyme reactions with the enzyme conjugated to a second antibody (i.e., dye as substrate and the chemiluminescence detection system) are presented and compared. This histoimmunoblot assay provides a general non-radioactive and sensitive immunohistochemical tool for the localization of compounds occurring in extracellular body fluid compartments. For inner ear research this method now enables the investigation of the penetration and distribution of therapeutics in peri- and endolymphatic sites and can even be applied to separately quantifying concentrations of a substance in different coils of the same cochlear section.

The Scala Vestibuli for Cochlear Implantation: An Anatomic Study

Traditionally, cochlear implantation has used the scala tympani (ST) for electrode insertion. When faced with ST ossification, the surgeon may elect to drill out the cochlea to accomplish partial electrode insertion. Theoretically, another option in this situation is to insert the electrode into the scala vestibuli (SV). To determine whether or not the dimensions of the SV are sufficient to accommodate an electrode array so as to assess the feasibility of SV cochlear implantation. The study of 20 normal human temporal bones, comparing the maximum diameter and surface area of the ST with those of the combined SV and scala media. The dimensions of the SV and scala media were comparable to those of the ST and appeared sufficient to accommodate a cochlear implant electrode array. It appears that the combination of SV and scala media is a viable alternative route for electrode insertion, at least on the basis of anatomic dimensions, in those cases in which the ST is obliterated.

Tetraethylammonium effects on cochlear potentials in the guinea pig

Voltage-dependent K + channels in the basolateral membrane of hair cells in guinea-pig cochlea might contribute to the non-linear current-voltage relationships in these hair cells and, thereby, to generation of the extracellular summating potential (SP). To evaluate the role of K + channels in the generation of the SP the perilymphatic perfusion technique was used to introduce the K +-channel blocker tetraethylammonium (TEA) into the cochlea. Sound-evoked cochlear potentials were measured subsequently. Without blocking nerve activity TEA induced reversible shifts of the SP in the negative direction, irrespective of whether we recorded from scala vestibuli or scala tympani. Shifts in the negative direction were probably due to TEA acting directly on the afferent fibres, since removal of nerve activity by the potent Na +-channel blocker tetrodotoxin (TTX) prevented TEA from shifting the SP in the negative direction. Once nerve activity had been removed by TTX, administration of TEA caused a small decrease in the magnitude of the SP, both in scala vestibuli and in scala tympani, irrespective of its polarity. The decrease was significant for the highest test frequencies only (8–12 kHz), and completely reversible. The rapidly activated K + channel in the inner hair cell (IHC) is probably blocked by TEA and this blocking might be responsible for the small decrease in magnitude of the SP. The asymmetric contribution from this K + channel to the IHC's current-voltage relationship seems to be only partly responsible for the generation of the SP, since blocking of this K + channel with TEA caused relatively small decreases in the amplitude of the SP. TEA did not affect the endocochlear potential.

Projections of the individual vestibular end-organs in the brain stem of the squirrel monkey

The central nervous system (CNS) projections of primary afferent neurons from individual vestibular receptors were studied using horseradish peroxidase (HRP) or biocytin labeling in 14 ears from 7 adult squirrel monkeys using the technique developed in the chinchilla (Lee et al., 1989, 1992). The specificity of labeling was verified by examining the location of the labeled fibers and cell bodies in the vestibular nerve and Scarpa's ganglion. Labeled fibers and cells were restricted to nerves and areas belonging to groups of cells in either the superior or the inferior ganglion of the vestibular nerve. In the vestibular nerve root, labeled primary afferent fibers also exhibited a receptor-dependent segregation at the entrance to the medulla. Fibers from the HSC and the SSC were found rostrally and those from the PSC and the SAC were found in the caudal area. The UTR fibers were situated intermediate between these two groups of fibers. (A bundle of fibers, probably vestibular efferents, was identified immediately rostrally and ventromedially to the UTR fibers.) The primary afferent fibers bifurcated into secondary ascending and descending fibers at the lateral border of the vestibular nuclei, forming a longitudinal rostrocaudal vestibular tract. The secondary fibers from individual end-organs occupied specific locations in the tract: the UTR fibers were dorsal to the SSC and the HSC fibers, PSC fibers were found most medially, and the SAC fibers occupied the lateralmost area. The secondary UTR fibers overlapped considerably with those of the SSC and the HSC. The orderly receptor-dependent segregation of fibers was more prominent in the descending tracts than in the ascending tracts. In the vestibular nuclei complex the location of the tertiary branches of various end-organs exhibited considerable overlap within the major vestibular nuclei (SN, superior nucleus; LN, lateral nucleus; MN, medial nucleus; DN, descending nucleus). There were still differences, however, in the projection pattern. Fibers from the SAC ran primarily in the lateral area, fibers from the SSC and the UTR were found ventromedially to the SAC fibers, and the HSC projected slightly medially to the fibers from the SSC. The PSC fibers projected most medially. The UTR and SAC sent numerous fibers to the cerebellum. Fibers from the semicircular canals projected through the rostrodorsal region of the SN and presumably also projected to the cerebellum. The precise termination of fibers was evaluated by studying the location of labeled boutons, which were identified in all major vestibular nuclei. Labeled boutons from all the receptors were in the rostral and central areas of the SN, and in the MN mainly in the rostral two-thirds. In the LN, boutons from all the receptors were in the rostroventral part, most of which were from the UTR and SAC. No labeled boutons were in the caudodorsal part of this nucleus. Labeled boutons in the DN primarily surrounded the descending tract fibers and were particularly prominent medially. In specimens in which superior vestibular nerve receptor organs were scratched vestibular efferent fibers were also labeled. These fibers traveled in the most ventral part of the vestibular nerve root and projected in the ventral aspect of the LN to labeled soma in the ipsilateral and contralateral brain stem. Specificity the in projection patterns of efferent fibers from different end-organs could not be ascertained.

Petrosal and Inner Ear of a Squalodontoid Whale: Implications for Evolution of Hearing in Odontocetes

ABSTRACT This study focuses on the petrosal and bony structures of the inner ear of a fossil squalodontoid whale (Odontoceti, Mammalia) from the Chandler Bridge Formation (late Oligocene) of South Carolina. High frequency hearing in toothed whales (odontocetes) has been attributed to specialized bony structures and their associated membranes in the inner ear cochlea. Whales also have very peculiar vestibule and semicircular canals by comparison to terrestrial mammals. By using serial grinding and computerized reconstruction, we identified in this fossil squalodontoid whale several inner ear structures specialized for high frequency hearing: 1) a well developed secondary bony lamina for the basilar membrane in the basal ¾ cochlear turn; 2) numerous foramina for the ganglion cells of the auditory nerve and a partially preserved spiral ganglion canal; 3) great size difference between the scala tympani and the scala vestibuli (including the scala media); and 4) wide separation between the cochlear turns. All these structures are correlated with high frequency hearing in modern odontocetes. The basilar membrane width, as inferred from the gap of the bony laminae preserved in the fossil, is greater in this squalodontoid than in modern delphinoids. The squalodontoid also has a higher cochlear spiral than modern delphinoids. These indicate that frequency of maximum sensitivity was lower in squalodontoids than in modern delphinoids. The vestibule of the squalodontoid is a very narrow and curved passage to the fenestra vestibuli. The semicircular canals are small and thin. These highly specialized structures suggest that specializations of the inner ear occurred prior to the diversification of all extant toothed whales, of which squalodontoids are the stem group.