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Hair cell distribution and orientation in goldfish otolith organs.

Structurally diverse sensory regions occur in the otolith organs of the goldfish inner ear. Scanning electron microscopy reveals regional distinctions based on three criteria. (1) Hair cells have different sizes of apical bundles, based on thickness. In all three maculae, two central regions have hair cells with bundles significantly thicker than those in surrounding regions. (2) Hair cell population density varies, with regional aggregations present. The central regions with thick bundles have two to three times the density of surrounding regions with thin bundles, and contain 40-80% of the total hair cell number in each macula. (3) Hair cell orientation maps show that each macula has two oppositely oriented cell populations that can be separated completely, not by a zone of interspersion, but apparently by a single unbroken line. The lagena is like the utricle in having hair cells with the kinocilium on the side of the cell toward the opposition line, but in the saccule the kinocilia face away from the line, and the small macula neglecta consists of two completely separate, oppositely oriented patches. The opposition line does not divide each macula simply down its midline; instead, the line divides the regions with thich bundles into nearly equal opposing areas, except for a remarkably abrupt large loop in the line in the anterior part of the saccule. The regional structural diversity in these organs may relate to localized functional diversity of responses to tilt, vibration and sound.

Animals

[Population of hair cells in the Corti's organ of shrews].

In shrews (family Soricidase) the organ of Corti was examined by means of the surface specimen technique. The analysis of the qualitative deviations from the normal cellular pattern of Corti organ was based on more than 160 ears of shrews (Sorex araneus, S. minutus, Neomys fodiens, and Crocidura suaveolens). The dislocation, rotation, malformation of hair cells, and one finding of the giant outer hair cell are described. The quantitative analyses of the population of hair cells are based on 61 ears of Sorex araneus divided into 2 age groups. The mean values of aplasias and of missing and supernumerary hair cells in dependence on individual rows and half-turns of cochlea are given. The loss of hair cells depends on the age of the animal. The findings of aplasias and supernumerary outer hair cells (especially those of atypical 4th row) are relatively frequent in shrews. The organ of Corti in shows has a very regular arrangement; deviations of all types are scant and thus we may hold the ideal cochleogram for the norm. The found deviations seem to have no functional meaning.

Age Factors

Properties of auditory nerve responses in absence of outer hair cells.

1. Recordings were made from chinchilla auditory nerve fibers after portions of the cochlear outer hair cell (OHC) population were destroyed with the antibiotic kanamycin. In most cases the inner hair cell (IHC) population was completely preserved as determined by phase-contrast microscopy. We presume that the remaining IHCs are functionally normal, and thus that recordings obtained from fibers originating from the lesioned cochlear segment reflect IHC behavior. 2. Behavioral thresholds were measured for all animals both before and after the production of the cochlear lesion. The audiograms and the histological evaluation of the ears were the basis for assessing whether a particular fiber originated in a normal, pathological (shifted threshold; IHC only), or border region. These criteria also identified the animals that sustained IHC damage together with the destruction of part of the OHC population. Only the data obtained from those fibers which probably originated from the OHC-free segment of the cochlea are considered in detail. 3. Fibers whose characteristic frequency (CF) identified them as belonging to the normal (audiometrically and histologically) region, were found to be normal in all respects. 4. Fibers from the border region (where the audiogram has a steep slope between normal and hearing-loss regions probably corresponding to the segment where OHC loss progresses from less than 10% to more than 90%) had very complex response patterns. Their frequency threshold curves (FTC) showed great variability. In general, the closer the fiber was to the fully developed lesion, the more abnormal its FTC became. 5. Those units that were concluded to have originated from the OHC-free part of the cochlea could be divided into three categories on the basis of the shape of their FTCs. A small fraction had very broad tuning (9%). The majority (53%) had approximately normal tail segment, normal bandwidth of the tip segment, and highly elevated threshold at CF. A group of fibers (38%) could not be assigned a CF. Probably the FTC of most of these latter fibers are similar to those of the previous group, but the sharply tuned short tip segment was either missed or was not reachable on account of its extremely high threshold level. 6. Such indexes of fiber response as latency, spontaneous rate, and time pattern (PST histograms) were not affected by the loss of OHCs. 7. On the basis of the data and of the assumptions made it was suggested that outer hair cells provide a frequency-dependent sensitizing influence to the inner hair cells. The frequency dependence could best be expressed as a flat-topped band pass characteristic.

Acoustic Stimulation

Genesis and maturation of vestibular hair cells.

The embryologic development in vivo and in vitro of mammalian hair cells in the crista ampullaris was continuously followed with regard to structural differentiation and maturation from the terminal mitosis to the morphological condition at partus or equivalent age in vitro. Otocysts were explanted both early and late during embryologic development: 13th and 16 gestation day, respectively. Both in fetal life and during in vitro conditions, the surface structures of the developing hair cell with regular arrangement of kinocilium/stereocilia were first differentiated, followed by a cytologic transformation intracellularly and subsequent development of nerve endings. Hair cells were able to develop without any morphologic contact with the nervous system. The afferent nerve system developed before the efferent nerve system (CBA/CBA mouse).

Animals

The coding of sound pressure and frequency in cochlear hair cells of the terrapin.

Intracellular recordings have been made from single hair cells in the cochlea of the terrapin, and the site of recording has been verified by injection of a fluorescent dye through the recording electrode. A hair cell gives periodic voltage responses graded with the intensity and frequency of the sound stimulus, and produces the largest response at its characteristic frequency. When small current steps are injected through the recording electrode, the voltage response of the cell exhibits damped oscillations at its characteristic frequency. The results are consistent with the idea that the cochlear frequency selectivity arises in two stages and it is suggested that the second stage resides within the hair cell itself.

Action Potentials

Intracellular studies of hair cells in the mammalian cochlea.

1. Intracellular recordings were made from inner hair cells in the first turn of the guinea-pig cochlea, the recording sites being confirmed by the injection of Procion yellow dye and subsequent histology. 2. The receptor potential, in response to a pure tone burst, consisted of an AC response which followed the wave form of the stimulus and was analogous to the extracellularly recorded cochlear microphonic and a depolarizating DC response which followed the envelope of the tone burst and was analogous to the extracellularly recorded summating potential. 3. The DC response was broadly tuned at high sound pressure having a maximal amplitude of 27 mV at a sound pressure level of ca. 100 db; however the bandwidth of the response was reduced at lower sound pressure level. Isoamplitude curves for the DC response were indistinguishable from the threshold curves for auditory nerve fibres. 4. The AC response was tuned in a similar fashion to the DC response except that it was attenuated at 6-9 db/octave with respect to the DC response. It is suggested that this difference was due to the effect of membrane capacitance and resistance on the AC response. In contrast the extracellularly recorded AC component was not subject to this attenuation. 5. The total resistance and capacitance in three cells were found to be 46-61 Momega and 7.8-15.8 muF respectively. 6. Intracellular resistance changes were measured during sound stimulation, the resistance change being proportional to the DC receptor potential, indicating constant current flow through the hair cell. The current varied between 0.37 and 0.81 nA between cells. The time constant for seven cells was found to lie between 0.31 and 0.76 msec. 7. A map of the basilar membrane showing position of hair cells against characteristic frequency corresponded to the cut-off frequencies of the basilar membrane mechanical measurements and the innervation sites of spiral ganglion cells.

Animals

Hair cell loss as a function of age in the normal cochlea of the guinea pig.

The surface specimen technique was used to study both spiral organs of 28 normal guinea pigs of four age groups: less than 24 hours, 6 weeks, 3 months and 1 year. Damaged hair cells were recorded for the whole of each spiral organ on cochleograms. The mean percentage number of outer hair cells damaged per age group was found to increase as a power function of age. In the animals aged less than 24 hours the mean percentage of damaged outer hair cells was 0.45%; in the 6-week animals, 1.85%; in the 3-month animals, 3.19%; and in the 1-year animals, 6.82%. At all ages outer hair cell loss was maximal in the third row, and towards the apex of the cochlea. Inner hair cell loss was very slight, with a maximum of 9 damaged inner hair cells per cochlea.

Aging

Psychophysical tuning curves and auditory thresholds after hair cell damage in the chinchilla.

Chinchillas were treated with kanamycin sulfate (150--200 mg/kg/day) to produce high-frequency hearing loss extending to about 4.0 kHz. Thresholds and psychophysical tuning curves (PTCs) were obtained before and after treatment, utilizing a shuttlebox avoidance procedure, and cochlear hair cells were evaluated under phase contrast microscopy. Hair cell loss resulting from kanamycin treatment varied from restricted lesions of the outer hair cells (OHCs) in the cochlear base, with no loss of inner hair cells (IHCs), to more extensive lesions involving both OHCs and IHCs. Threshold shift of at least 40 dB was always associated with OHC loss. PTCs obtained from frequency regions exhibiting 40--50 dB of threshold shift were normal in shape. With threshold shift in excess of 50 dB, PTCs were progressively distorted, with truncation of the tip segment and in some cases increased sensitivity of the tail segment. The results suggest that the threshold of optimally functional IHCs after kanamycin-induced OHC loss is about 40 dB higher than normal. Threshold shift in excess of 40 dB may represent IHC damage. IHCs are capable of transducing the fine-frequency information necessary for generating normally sharp PTCs in the absence of OHCs. However, with threshold shift in excess of approximately 50 dB, this frequency resolution is increasingly compromised.

Animals

Synaptic structures in the type II hair cell in the vestibular system of the guinea pig. A freeze-fracture and TEM study.

The synaptic contacts of the type II hair cell in the vestibular system of the guinea pig was described in thin-sectioned and freeze-fractured specimens. Synaptic bodies were present at the apposition with both large and small afferent terminals. About 20% of the synaptic bodies observed consisted of complexes of two or more adjacent synaptic discs. In freeze-fracture replicas, the cytoplasmic leaflet of the hair cell plamalemma beneath the synaptic body had a bar-shaped aggregate of large particles. The size and shape of the particle aggregate was the same as that of the synaptic body. Small plasmalemmal deformations, interpreted as sites of synaptic vesicle exocytosis, were found immediately adjacent to the particle aggregate. On the postsynaptic membrane, an aggregate of intramembrane particles was present at the synaptic junction. The type II hair cell had no gap junctions or close membrane appositions between it an the apposed afferent fiber. Efferent boutons ending on the type II hair cell had no intramembrane particle specialization on the postsynaptic membrane; however those efferent boutons ending on large and small afferent fibers had an aggregate of medium-sized particles on the external leaflet of the postsynaptic bouton beneath the presynaptic active zone.

Animals

Hearing and hair cells.

Animal experiments and human temporal bone studies have indicated that hearing losses may occur in ears that have normal populations of sensory cells. The possibility exists that the hair cells have been rendered non-functional because of ultrastructural damage. Preliminary investigation of cilia on hair cells in areas of suspected damage shows no difference from cilia in similar areas of control ears.

Adolescent

Permanent threshold shift and cochlear hair cell loss in the kanamycin-treated guinea pig.

The differential contribution of the inner hair cells (IHC) and the outer hair cells (OHC) in the mammalian cochlea to hearing sensitivity was assessed in six behaviorally-trained guinea pigs by comparing audiograms preadministration and postadministration of kanamycin, an antibiotic that predominantly destroys guinea pig OHC while leaving the IHC structurally unchanged. The results support the hypothesis that only the IHC of the cochlea responds to tones approximately 50 to 60 dB above the threshold of the intact cochlea.

Animals

Further studies on the Schroeder-Hall hair-cell model.

The Schroeder-Hall hair-cell model [M.R. Shroeder and J.L. Hall, "Model for mechanical to neural transduction in the auditory receptor," J. Acoust. Soc. Am. 55, 1055-1060 (1974] was further explored using additional stimulus waveforms and analysis techniques. The model is shown to have other interesting properties such as the ability to generate realistic two-tone interactions. Amended to limit the growth of transmitter release, the model also produces realistic adaptation and incremental response data. Other amendments improved high-intensity period histogram waveshapes. No one amendment, however, allowed the model to produce both realistic adaptation curves as well as period histogram waveforms that faithfully mimicked physiological data.

Hair Cells, Auditory

CASZ1 regulates the maturation of outer hair cells and is required for hearing in mice.

The transcription factor ATOH1 is a master regulator of mechanosensory hair-cell (HC) development in the ear. Here, we report that its target gene Casz1 regulates the maturation of outer HCs (OHCs). Genetic deletion of Casz1 during (but not after) cochlear development in the mouse caused: hearing loss; disorganization of mechanosensory stereocilia bundles in OHCs; reduced F-actin density in OHC cuticular plates; progressive OHC loss; and mild morphological alterations in inner HCs. This deletion also altered gene expression, delaying downregulation of genes expressed in immature OHCs, including the actin regulator-encoding gene Coro2a, and accelerating upregulation of genes expressed in mature OHCs. Deleting Coro2a in Casz1 mutant mice restored F-actin density in cuticular plates but increased stereocilia bundle disorganization and hearing thresholds, revealing that CORO2A provides an overall beneficial effect. Our data indicate that CASZ1 regulates transcriptional and morphological maturation of OHCs, and that CASZ1 in maturing HCs is necessary for hearing.

CASZ1

Inhibition by efferent nerve fibres: action on hair cells and afferent synaptic transmission in the lateral line canal organ of the burbot Lota lota.

1. Intracellular recordings were made from morphologically identified hair cells in the lateral line canal organs of the burbot Lota lota. 2. I.p.s.p.s were recorded from hair cells when the efferent fibres were excited by electrical stimulation of the lateral line nerve. The i.p.s.p.s were abolished when the fish was injected with immobilizing concentration of Flaxedil which is known to block the efferent synapses. 3. The i.p.s.p.s are accompanied by a decrease in the resistance of the hair cell membrane and an increase in the intracellular receptor potential. 4. Spontaneous and mechanically evoked e.p.s.p.s which were recorded intracellularly from the post-synaptic afferent nerve terminals were reduced in amplitude for the duration of the i.p.s.p.

Animals

Hair cell degeneration in guinea pigs intoxicated with kanamycin during intrauterine life; a structural and ultrastructural study.

The structural ototoxic effect of kanamycin during intrauterine life has been shown in guinea pigs. The extent of the loss mostly affected the outer hair cells of the basal part of the cochlea. Ultrastructural studies demonstrated a hair cell degeneration pattern similar to that previously described in adult animals following antibiotic intoxication.

Animals

Extra internal hair cells. A scanning electron microscopic study.

The extra internal hair cell (EIHC) of the human cochlea was observed by means of a scanning electron microscope. The EIHC was found not infrequently in all turns of the human cochlea. It was located medial to the IHC row. The inner pillar cells showed an abnormal structure. The anatomical relationships between the displaced IHC and EIHC, and the inner pillar cell were classified into five types. The origin of these anomalies is discussed form an embryological viewpoint.

Aged

Pathological actin in vestibular hair cells of the waltzing guinea pig.

Vestibular type 1 hair cells in the waltzing guinea pig contain needle-shaped inclusion bodies which grow in an uncontrolled fashion associated with the destruction of the cell. The needles are shown to be composed by filaments of actin, a protein identified in the electron microscope by its ability to bind subfragment 1 of myosin. Whereas actin filaments in stereocilia are oriented down towards the cell body, filaments in the needles point up towards the cuticular plate. The hereditary lesion appears to be associated with a defective control of polymerization of actin into filaments.

Actins