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C M Hackney

Publications and source records attributed to C M Hackney.

At least 37 records · Page 2Linked to original sources
PubMed1993

The functional morphology of stereociliary bundles on turtle cochlear hair cells.

The stereociliary bundles of hair cells from the basilar papilla of the red-eared turtle were examined with transmission and high resolution scanning electron microscopy to provide a description of their morphology, orientation and inter-ciliary connections for comparison with physiological observations. Bundles on hair cells in the basilar membrane region are of a uniform shape and orientation, but bundles on the apical and basal limbus are distinct in having elongated kinocilia more than twice the length of the tallest stereocilia. Bundles in the basilar membrane region show a roughly two-fold increase in height from 5 to 9 microns from base to apex. Electrical recordings from isolated hair cells indicate that the bundle height is inversely proportional to the cell's characteristic frequency. It is argued that the change in dimensions is insufficient to contribute significantly to the cochlea's frequency selectivity. The cytoplasm adjacent to the kinocilium is filled with microtubules and large vesicles, and there are coated pits in the apical membrane which, it is suggested, may be indicative of rapid turnover of the membrane in this region.

Animals↗
PubMed1993

Microslicing of the resin-embedded cochlea in comparison with the surface preparation technique for analysis of hair cell number and morphology.

The surface preparation technique for hair cell counting is frequently used for the analysis of cochlear pathology in relation to hearing impairment. To overcome problems encountered with standard surface preparation methods, a new combination of resin embedding and microslicing has been applied to the mammalian cochlea, to permit hair cell counting to be followed by electron microscopic analysis. Partial removal of the cochlear shell is followed by resin infiltration under vacuum prior to polymerization. The cochlea is then subdivided into hemicoils using a mounted annular diamond blade. Embedding before microslicing ensures that both apical and basal regions of the cochlea are preserved equally well with an evenly distributed and therefore predictable 10% hair cell loss. By comparison, the standard surface preparation method often produces distortion of remaining organ of Corti and unpredictable losses of hair cells, for example, up to 17%. The damage tends to be greatest towards the base of the cochlea, an area which shows particular susceptibility to ototoxic agents such as the aminoglycoside antibiotics. Thus, for assessment of pathology caused by ototoxic agents, this method has considerable advantages over the surface preparation technique.

Animals↗
PubMed1992

Putative immunolocalization of the mechanoelectrical transduction channels in mammalian cochlear hair cells.

Hair cells bear an apical bundle of stereocilia arranged in serried rows. Deflection of the bundle controls the opening and closing of mechanoelectrical transduction channels, thereby altering the conductance across the apical plasma membrane. Two locations for these channels have been proposed in the bundle, either near the bases of the stereocilia or towards their tips. One hypothesis that is consistent with the latter possibility suggests that fine extracellular filaments, which run between the tips of the shorter stereocilia and the sides of the taller stereocilia behind, operate the channels. Determining the precise position of the channels is essential to test this hypothesis. We have therefore attempted to localize them immunocytochemically. Because hair-cell transduction is amiloride sensitive, the channels may have an amiloride-binding site associated with them. We have therefore used a polyclonal antibody raised against another amiloride-sensitive ion channel to hunt for them. This antibody recognizes a 62-64 kDa band in immunoblots of cochlear tissue, and produces discrete labelling in the hair bundle. This is most concentrated just below the tips of the shorter stereocilia, coinciding with a region of specialization in the closely apposed membranes of the short and tall stereocilia but not with either end of the tip link.

Amiloride↗
PubMed1992

An atlas of glycine- and GABA-like immunoreactivity and colocalization in the cochlear nuclear complex of the guinea pig.

The distribution and colocalization of gamma-aminobutyric acid (GABA)- and glycine-like immunoreactivity in the cochlear nuclear complex of the guinea pig have been studied to produce a light microscopic atlas. The method used was based on post-embedding immunocytochemistry in pairs of 0.5-micron-thick plastic sections treated with polyclonal antibodies against conjugated GABA and glycine respectively. Immunoreactive cells, presumably short axon neurones, predominated in the dorsal cochlear nucleus, with mostly single-GABA-labelled cells in the superficial layer, double-labelled in the middle, and single-glycine-labelled in the deep layers. A few large single-glycine-labelled cells, interpreted as commissural neurons, occurred in the ventral nucleus. Scattered double-labelled cells, probably Golgi cells, were seen in the granule cell domain. Immunolabeled puncta of all three staining categories occurred in large numbers throughout the complex, apposed to somata and in the neuropil, showing a differential distribution onto different types of neuron. Three immunolabeled tracts were noted: the tuberculoventral tract, the commissural acoustic stria, and the trapezoidal descending fibres. Most of the fibres in these tracts were single-labelled for glycine, although in the last mentioned tract single-GABA- and double-labelled fibres were also found. Some of the immunolabeled cell types described here are proposed as the origins of the similarly labelled puncta and fibres on the basis of known intrinsic connections.

Animals↗
PubMed1992

Behavioural evidence for recovery of auditory function in guinea pigs following kanamycin administration.

Deterioration followed by recovery of behavioural absolute threshold and frequency selectivity has been observed in guinea pigs following kanamycin administration of 200 mg/kg body weight daily for 16 days. Deterioration in function consistently follows a high-to-low frequency pattern and recovery generally occurs at the lowest of the high (8-32 kHz) frequencies affected. The degree of recovery is related to the magnitude of the threshold elevation; where large (40-45 dB) elevations occur initially, the process appears to be partial since threshold recovers only to within 5-12 dB of pre-administration levels. In instances where smaller threshold elevations (5-20 dB) take place initially, recovery can sometimes be complete. However, when threshold elevations of over 50 dB occur, no recovery is apparent. Recovery is relatively slow, taking place over periods of up to 100 days post-kanamycin administration. Hair cell counts have established that the threshold elevation which remains in instances of partial recovery is not related to a reduction in hair cell numbers at the light microscope level.

Acoustic Stimulation↗
PubMed1991

Localisation of putative mechanoelectrical transducer channels in cochlear hair cells by immunoelectron microscopy.

Displacement of the apical stereociliary bundle of cochlear hair cells mechanically gates transducer channels. Knowing the position of the channels with regard to the apical structures of the hair cell could indicate how this mechanism operates. At present, there is conflicting evidence regarding their precise location; the channels have been suggested to be located either towards the base of the stereocilia or at the tips where they could be operated by extracellular links running from the top of shorter stereocilia to the sides of adjacent taller ones. The channels have been shown to be reversibly blocked by amiloride. This has prompted us to use a polyclonal antibody raised against another amiloride-sensitive channel to search for them using immunolabelling. The location of the primary antibody has been revealed using pre-embedding labelling with a colloidal gold-conjugated secondary antibody followed by scanning transmission electron microscopy of semi-thin sections. In this way, more complete information on the relationship of the labelling to the three-dimensional organisation of the stereociliary bundle has been obtained in comparison with previous immunofluorescence and transmission electron microscopic results. Labelling occurs in discrete areas towards the tips of the stereocilia, one of the possible sites for the transducer channels, predominantly between the membranes of shorter and taller stereocilia.

Amiloride↗
PubMed1990

Organization of microtubules in cochlear hair cells.

The organization of microtubules in hair cells of the guinea-pig cochlea has been investigated using transmission electron microscopy and correlated with the location of tubulin-associated immunofluorescence in surface preparations of the organ of Corti. Results from both techniques reveal consistent distributions of microtubules in inner and outer hair cells. In the inner hair cells, microtubules are most concentrated in the apex. Reconstruction from serial sections shows three main groups: firstly, in channels through the cuticular plate and in a discontinuous belt around its upper perimeter; secondly, forming a ring inside a rim extending down from the lower perimeter of the plate; and thirdly, in a meshwork underlying the main body of the plate. In the cell body, microtubules line the inner face of the subsurface cistern and extend longitudinally through a tubulo-vesicular track between the apex and base. In outer hair cells, the pattern of microtubules associated with the cuticular plate is similar, although there are fewer present than in inner hair cells. In outer hair cells from the apex of the cochlea, microtubules occur around an infracuticular protrusion of cuticular plate material. In the cell body, many more microtubules occur in the region below the nucleus compared with inner hair cells. The possible functions of microtubules in hair cells are discussed by comparison with those found in other systems. These include morphogenesis and maintenance of cell shape; intracellular transport, e.g., of neurotransmitter vesicles; providing a possible substrate for motility; mechanical support of structures associated with sensory transduction.

Animals↗
PubMed1990

Anatomy of the cochlear nuclear complex of guinea pig.

The cyto- and fibre-architecture of the cochlear nuclear complex of the guinea-pig has been studied in serial sections using Nissl, Golgi and combined cell-myelin staining of normal material, and a silver degeneration method after cochlear ablation. The nuclear subdivisions and major cell types can be recognised on the basis of those found in the cat, but there are some differences between the two species in the precise distribution and morphology of the neurons. The rostrodorsal part of the anteroventral cochlear nucleus (AVCN) contains predominantly spherical bushy cells, but these cannot be readily divided into large and small types as in the cat. Globular bushy cells are seen in the caudal region of the AVCN, but the majority occur in the posteroventral cochlear nucleus (PVCN), in an area extending from the nerve root right up to the boundary of the dorsal cochlear nucleus (DCN). The octopus cells constitute a distinct region in the most dorsomedial part of the PVCN underneath the DCN. Giant cells are seen scattered around the nerve root region. Multipolar and small cells are seen throughout the non-granular regions of the ventral cochlear nucleus (VCN) except for the octopus cell area, but occur mainly in the more rostral regions of the PVCN. Small cells occur in greatest abundance in the thin cap area at the dorsal edge of the VCN below a superficial granule cell layer. The latter covers the dorsolateral surface of the VCN, and a lamina of granule cells partially separates the PVCN from the DCN. The DCN can be divided into four layers. The outermost molecular layer (layer 1) is separated from the deeper regions by a prominent layer of granule cells (layer 2) which also contains the pyramidal cells. Molecular layer stellate cells are seen in layer 1 and a staggered row of cartwheel neurons is found at the boundary between layers 1 and 2. Layer 3 contains the basal dendrites of the pyramidal cells and some small (vertical) cells, and is innervated by the descending branches of the cochlear nerve. The deepest layer 4, which contains multipolar cells and giant cells, does not appear to receive this direct cochlear input.

Animals↗
PubMed1990

Comparative ultrastructure of subsurface cisternae in inner and outer hair cells of the guinea pig cochlea.

The subsurface cisternal systems of outer hair cells (OHCs) from different cochlear regions have been compared with the subsurface cisternal system of inner hair cells. Three main observations have been made: (1) the number of cisternal layers, when there is more than one present, is reduced along the length of an individual outer hair cell; (2) basal outer hair cells may have only one fenestrated cisternal layer; and (3) the inner hair cells possess a lateral cistern and associated pillar and filament complexes which are very similar to those of some basal OHCs. These observations are discussed in relation to hypotheses regarding the role of these structures in hair cell motility.

Animals↗
PubMed1989

Tubulin and microtubules in cochlear hair cells: comparative immunocytochemistry and ultrastructure.

The distribution of tubulin has been investigated in surface preparations of the guinea pig organ of Corti using indirect immunofluorescence microscopy. Two different monoclonal antibodies to tubulin produce similar distinct patterns of labelling in hair cells. Labelling is greater in inner hair cells than outer hair cells. It occurs in rings around the cell apex, and in a meshwork below and channels through, the cuticular plate. In outer hair cells from the apical region of the cochlea, labelling occurs around the location of a basalward protrusion of the cuticular plate. These patterns correlate with the location of microtubules observed using transmission electron microscopy. A large patch of labelling occurs on the strial side of the cell corresponding to the largest channel through the cuticular plate and the kinociliary basal body. Strands of labelling are seen running parallel to the long axis of the cell between the subcuticular and synaptic region. Many more of these strands are seen in the inner hair cell than the outer hair cell and may correspond to tracks of microtubules transporting neurotransmitter vesicles or other organelles. In outer hair cells, intense labelling and many microtubules are seen in the subnuclear region. The possible roles of the different microtubule arrangements are discussed.

Animals↗
PubMed1988

Stereociliary cross-links between adjacent inner hair cells.

An extensive network of intracellular cross-links occurs between the stereocilia of each cochlear hair cell bundle. These links fall into two main categories; lateral links which run roughly horizontal with respect to the reticular lamina and which join stereocilia of the same or adjacent rows, and tip-to-side links which run at a more vertical angle from the tip of each shorter stereocilium to the side of the adjacent longer stereocilium in the row behind. It has been proposed that deformation of the tip-to-side links causes alteration of the rate of opening of ion channels, producing transduction. Lateral linkages also occur between the stereocilia of adjacent hair cells. Now, intercellular links which resemble the tip-to-side links have been observed. Some of these occur in positions inappropriate to their proposed role in transduction. Several hypotheses are proposed to account for their presence e.g., the links could represent the remnants of a glycocalyx which is best preserved in areas where stereocilia are closely opposed.

Animals↗
PubMed1986

Intercellular cross-linkages between the stereociliary bundles of adjacent hair cells in the guinea pig cochlea.

Hair cells of the guinea pig organ of Corti have been examined using high resolution scanning electron microscopy. In addition to the extensive array of cross-links between the stereocilia of individual hair cells which have been reported previously, we have seen examples of attachments between the stereocilia of both adjacent inner and adjacent outer hair cells. The implications of these observations are discussed.

Animals↗
PubMed1986

High-resolution scanning-electron microscopy of stereocilia using the osmium-thiocarbohydrazide coating technique.

Further observations on the detailed morphology of stereocilia have been made using high-resolution scanning-electron microscopy of osmium-thiocarbohydrazide-coated guinea pig cochleae. Three types of cross-link have been observed between stereocilia. Side-to-side and row-to-row linkages are composed of a filamentous network whilst upward-pointing links are a fine single strand, often with a terminal widening. The stereocilia have rough surfaces. These features are observed on both inner and outer hair cells despite reported sensitivity to long periods of osmium fixation. We suggest that osmium sensitivity may be altered by the buffering conditions used during preparation. The observations on osmium-coated material correspond more closely with those made using transmission-electron microscopy than those made using other scanning-electron microscopical preparation techniques, since gold-coating artefacts are absent and the degree of specimen collapse is less. This has enabled us to observe fine details of the links and their attachments which have not been reported previously in SEM.

Animals↗
PubMed1986

The distribution of spherical cells in the anteroventral cochlear nucleus of the guinea pig.

Several types of neuron are found in Nissl-stained sections of the anteroventral cochlear nucleus (AVCN). From these one group, the spherical cells (Osen, 1969), can be readily distinguished from the remaining small and multipolar forms. The rostral pole of the AVCN has previously been subdivided into the large and the small spherical cell areas (in several mammals). In the present study of the guinea pig AVCN, spatial distributions of cell density, size, and shape have been investigated. These have been used to test whether the subdivision made on the basis of morphological differences in the spherical cells is valid, or whether there is a gradual gradient in these features. This analysis has shown that although variations in cell size and shape are observed, the spherical cell area cannot be partitioned on these grounds. There is, however, a graded increase in spherical cell packing density towards the rostral pole of the AVCN, with proportionately fewer of the other cell types present.

Animals↗
PubMed1986

Morphological changes to the stereociliary bundles in the guinea pig cochlea after kanamycin treatment.

Pigmented guinea pigs were treated with 400 mg/kg of kanamycin for 8 days and sacrificed 13 days later. The upper surface structures of the organ of Corti were studied using high resolution scanning electron microscopy to reveal fine details of stereociliary fusion and damage to the reticular lamina. Outer hair cell stereocilia showed from partial to complete fusion and loss whilst the inner hair cells showed very little evidence of any damage. In particular, the possible effects of kanamycin on the cross-links between the stereocilia have been investigated. The cross-links are found to be present on all outer and inner hair cell bundles which show no fusion of stereocilia. Partially-fused stereocilia which have free tips still possess the upward-pointing links, and the remaining undamaged stereocilia in these bundles possess the normal cross-links. Where fusion occurs along the sides of the stereocilia, the side-to-side links are missing although whether their absence either results in, or else is caused by, the fusion is not known.

Animals↗
PubMed1985

Cross-links between stereocilia in the guinea pig cochlea.

Cross-links between stereocilia on guinea pig cochlear hair cells have been examined using high resolution scanning (SEM) and transmission electron microscopy (TEM), confirming recent descriptions of these structures. Links from the tips of shorter stereocilia to the sides of the adjacent taller stereocilia (upward-pointing links), between stereocilia of the same row (side-to-side links) and between adjacent rows (row-to-row links), have been observed on inner and outer hair cells. These links have been seen in material fixed using (1) glutaraldehyde only, (2) glutaraldehyde/osmium and (3) glutaraldehyde/osmium/thiocarbohydrazide (a technique which makes gold coating unnecessary). Upward-pointing links were seen less frequently, and the surfaces of stereocilia and microvilli were smoother after fixation (3) compared with fixations (1) and (2) in which they were usually roughened in appearance. In TEM, side-to-side and row-to-row links form a regular lattice between stereocilia, and consist of a number of strands. Upward-pointing links consist of a single strand, the ends of which are associated with electron-dense material. This lies between the stereociliary membrane and the actin filament bundle, at the tip of the shorter stereocilium and the side of the taller stereocilium.

Animals↗