Biomedical subjects

S D Comis

Publications and source records attributed to S D Comis.

At least 19 recordsLinked to original sources

PubMed1999

Hair cell loss in the aged guinea pig cochlea.

The various effects of ageing on the auditory system, collectively termed presbycusis, are being studied across a wide range of animal species, including humans. One contributing factor to presbycusis is thought to be losses of the sensory hair cells in the cochlea. In this study, hair cell counts were obtained from cochleas of pigmented guinea pigs (Cavia porcellus) at ages ranging from 11 days to 4 years 7 months, using scanning electron microscopy to visualize the organ of Corti. Representative samples of the basal, middle and apical turn of the cochlea were photographed for analysis. Hair cell loss was observed, even in young animals. However, the loss was greater in the aged animals, but was not distributed evenly throughout the length of the cochlea. No significant loss of hair cells was seen in the basal (high frequency) or middle turn of the cochlea of the aged animals. In the apical (low frequency) turn, there was a significant loss of hair cells in all rows of outer hair cells (up to around 20%), and was most severe in the third row. There was no loss of apical inner hair cells in the aged animals.

Aging↗

PubMed1998

Ototoxicity resulting from intracochlear perfusion of Streptococcus pneumoniae in the guinea pig is modified by cefotaxime or amoxycillin pretreatment.

Acute changes in the electrophysiology and ultrastructure of the organ of Corti were studied after microperfusion of c. 5 x 10(6) CFU of serotype 2 Streptococcus pneumoniae D39 or Escherichia coli K-12 directly into the scala tympani of guinea pigs. Hearing loss was assessed by recording the auditory nerve compound action potential response to a 10 kHz tone pip. Mean hearing loss 3 h after pneumococcal perfusion (n = 4) was 44 dB, compared to 6 dB after E. coli perfusion (n = 4) (P<0.001). After pneumococcal perfusion, scanning electron microscopy revealed damage to hair cell stereocilia and cratering of the apical surface of supporting cells. Intraperitoneal injection of 100 mg/kg cefotaxime (n = 4) or 100 mg/kg amoxycillin (n = 4) 30 min before perfusion of pneumococci significantly reduced mean hearing loss to 23 dB (P=0.01) or 20 dB (P=0.01), respectively, and diminished ultrastructural damage. The data suggest that if pneumococci invade the inner ear during meningitis, cochlear deafness may rapidly ensue.

Amoxicillin↗

PubMed1998

The auditory brainstem response of aged guinea pigs.

The auditory brainstem response (ABR) technique was used to investigate potential dysfunctions in the auditory brainstem of the pigmented guinea pig (Cavia porcellus) associated with biological ageing. Animals aged from 58 days to 4 years 3 months were tested. ABRs were recorded at stimulation intensities from 85 dB HL to -10 dB HL. The auditory thresholds were found to undergo marked elevations in old animals, by an average of 32 dB. From the traces obtained, four positive deflection waves were reliably recorded. The latency of each of the four waves was evaluated at different stimulation intensities in guinea pigs of different ages. Although there was a trend for the latencies to increase in old age, these differences were not statistically significant. Similarly, there were no significant age-related changes in the inter-peak intervals. The latency/intensity functions of the four waves produced parallel curves. However, the curve from the old age group was shifted to the right, by an average of 35 dB, indicative of conductive hearing loss. There was no evidence of retro-cochlear hearing loss. Therefore, it appears that the threshold elevations in the old animals can be accounted for by conductive hearing loss, presumably in the middle ear. In 24% of the old animals tested, no ABR could be elicited. It would appear that these animals had suffered severe sensorineural hearing loss.

Aging↗

PubMed1997

A role for pneumolysin but not neuraminidase in the hearing loss and cochlear damage induced by experimental pneumococcal meningitis in guinea pigs.

We investigated the roles of pneumolysin and neuraminidase in the pathogenesis of deafness and cochlear damage during experimental pneumococcal meningitis. Anesthetized guinea pigs were inoculated intracranially with 7.5 log10 CFU of either (i) wild-type Streptococcus pneumoniae D39 (n = 8), (ii) PLN-A, a defined isogenic derivative of D39 deficient in pneumolysin (n = 5), or (iii) deltaNA1, a new derivative of D39 deficient in neuraminidase constructed by insertion-duplication mutagenesis of the nanA gene (n = 5). To quantify hearing loss, the auditory nerve compound action potential evoked by a tone pulse was recorded from the round window membrane of the cochlea every 3 h for 12 h. The organ of Corti was intravitally fixed for subsequent examination by high-resolution scanning and transmission electron microscopy. All animals sustained similar meningeal inflammatory responses. PLN-A induced significantly less hearing loss than D39 over the frequency range of 3 to 10 kHz. Levels of mean hearing loss at 10 kHz 12 h postinoculation were as follows: D39, 50 dB; deltaNA1, 52 dB (P = 0.76 versus D39), and PLN-A, 12 dB (P < 0.0001 versus D39). The mean rates of hearing loss at 10 kHz were 4.4 dB/h for D39, 4.3 dB/h for deltaNA1, and just 1.0 dB/h for PLN-A (P < 0.0001 versus D39). Suppurative labyrinthitis was universal. PLN-A induced the accumulation of less protein in the cerebrospinal fluid (P = 0.04 versus D39). Infection with D39 and deltaNA1 induced significant damage to the reticular lamina, the sensory hair cells, and supporting cells of the organ of Corti. By contrast, after infection with PLN-A, the organ of Corti appeared virtually intact. Pneumolysin seems to be the principal cause of cochlear damage in this model of meningogenic deafness. No clear pathogenic role was demonstrated for neuraminidase.

Animals↗

PubMed1997

Possible involvement of nitric oxide in the sensorineural hearing loss of bacterial meningitis.

Microperfusion of scala tympani with the NO donors, sodium nitroprusside (SNP) and S-nitroso-N-acetylpenicillamine (SNAP), produced marked depression of the compound action potential (CAP) and cochlear microphonic (CM) together with severe and widespread morphological damage to hair cells and supporting cells in the organ of Corti. In addition, direct perfusion of N-methyl-D-aspartate (NMDA) into scala tympani, which probably induces excess stimulation of NMDA receptors within the cochlea and which is known to lead to the release of NO, was found to elicit similar electrophysiological and structural lesions in the cochlea. Pre-perfusion of scala tympani with L-methyl arginine (L-MA), which inhibits the release of NO, or superoxide dismutase (SOD), an O2-scavenger, conferred marked protection upon the cochlea from the lesions caused by NO donors. These observations indicate that enhanced NO production is likely to be an important factor responsible for pathological insult of the cochlea. The possibility is discussed that this factor is involved in the chain of events leading to hearing loss caused by bacterial meningitis. Such hearing loss is a major sequela of bacterial meningitis in children.

Action Potentials↗

PubMed1995

NG-methyl-L-arginine protects the guinea pig cochlea from the cytotoxic effects of pneumolysin.

Sensorineural hearing loss is a major sequela of the bacterial meningitis associated in particular with Streptococcus pneumoniae. Recent studies have shown pneumolysin, a toxin elaborated by S. pneumoniae, to be cytotoxic to the guinea pig cochlea. The mechanisms of this cytotoxicity are, however, not fully understood. In the present study this deleterious action of pneumolysin has been shown to be blocked by pretreating the cochlea with NG-methyl-L-arginine, a known inhibitor of nitric oxide synthesis. Furthermore, pretreatment of the cochlea with MK-801, an NMDA receptor antagonist, was also found to confer marked protection from the action of pneumolysin. This latter finding is consistent with previous reports that excess stimulation of NMDA receptors within the cochlea, an event known to lead to excess nitric oxide release, have similar effects on the cochlea as pneumolysin perfusion. It would therefore appear that nitric oxide may represent a significant link in the chain of events leading to the deafness of bacterial meningitis.

Action Potentials↗

PubMed1995

The cochlear lesion in experimental bacterial meningitis of the rabbit.

Sensorineural hearing loss was studied in a rabbit model of experimental bacterial meningitis using electrophysiological and ultrastructural techniques. Hearing impairment was monitored by auditory brain-stem evoked responses (ABERs) and concomitant structural lesions were identified by both transmission (TEM) and scanning (SEM) electron microscopy. Meningitis was induced by intra-cerebrospinal fluid injection of either Escherichia coli (strain 2073 and type K-12) or Haemophilus influenzae type b. Auditory loss of approximately equal to 10 dB occurred in all rabbits by about 10 hours post infection and progressed in severity until by 20 h following infection, hearing losses up to and > 60 dB were obtained. At levels of hearing loss < 20 dB ultrastructural damage to the organ of Corti was barely detectable. With greater levels of hearing loss, patchy structural damage to hair cells, synaptic nerve terminals, supporting cells and inner spiral sulcus cells and cells of the stria vascularis was clearly evident. Bacteria were found in scala tympani, the basilar membrane, the organ of Corti, scala media, the spiral ligament and at the margin of the stria vascularis. Evidence of bleeding was found in some cochleas; erythrocytes were found in scala tympani, scala media, amongst hair cells and beneath the tectorial membrane. The results show that hearing loss is associated with bacterial invasion and damage to the organ of Corti and that the cause of hearing loss is likely to result from multiple lesions within the cochlea. Lesions to sensory cells almost certainly will produce permanent hearing loss. Lesions to supporting cells, nerve terminals and to stria vascularis may well produce only temporary hearing loss.

Animals↗

PubMed1993

Ependymal cells of the choroid plexus express tumour necrosis factor-alpha.

Tumour necrosis factor-alpha (TNF alpha) is a major proinflammatory cytokine which appears in the cerebrospinal fluid very early after endotoxin challenge, and is likely to be produced locally. Following in vivo and in vitro challenge with endotoxin, we have demonstrated immunocytochemically and by in situ hybridization that pig and guinea-pig choroid plexus ependymal cells can produce TNF alpha. Immuno-electron microscopy shows that this protein is localized within ependymal cells to the cytoplasm and microvilli. We suggest that this TNF alpha may be important in the initiation of the inflammatory response in bacterial meningitis.

Animals↗

PubMed1993

Cytotoxic effects on hair cells of guinea pig cochlea produced by pneumolysin, the thiol activated toxin of Streptococcus pneumoniae.

The cytolytic toxin, pneumolysin, from the gram positive bacterium, Streptococcus pneumoniae, when perfused through the scala tympani of the guinea pig cochlea reduced the amplitude of both the compound action potential and the cochlear microphonic potential. When the surface of the organ of Corti was examined by scanning electron microscopy, both inner and outer hair cells and supporting cells were found to be damaged. Inner hair cells and outer hair cells of row 3 were the most susceptible to damage by pneumolysin, followed by row 2 and then by row 1 of the outer hair cells. Damage to hair cells included disruption and splaying of stereocilia, loss of stereocilia and complete dissolution of hair bundles. Apical surfaces of hair cells and supporting cells were torn, pitted and cratered with shrinkage and tearing of cell boundaries. Within the dose range perfused (0.05-1 micrograms/microliters in a 10 microliters aliquot), the magnitude of the physiological and anatomical lesions was concentration dependent. The cytotoxic effects of pneumolysin reported here may be clinically significant factors in deafness caused by meningitis and otitis media in humans.

Animals↗

PubMed1991

Endotoxin induced damage to the cochlea in guinea pigs.

An apparently unique form of cochlear damage was produced in guinea pigs by perfusing the cochlea or injecting the cerebrospinal fluid with bacterial endotoxin. This developed rapidly (within two hours) and was characterised by swelling of the tectorial membrane and damage to both inner and outer hair cells, with parallel functional damage demonstrable electrophysiologically. All these changes could be attenuated by pretreatment with dexamethasone. Such endotoxin mediated lesions may be the mechanism by which hearing loss occurs in bacterial meningitis.

Action Potentials↗

PubMed1991

The effect of bacterial endotoxin upon the morphology of the tectorial membrane and stereocilia in the guinea pig cochlea.

Endotoxin of E coli was microperfused into scala tympani or injected into the cerebrospinal fluid in anaesthetised pigmented guinea pigs. The effects of endotoxin on the cochlea were studied using electrophysiological techniques and scanning electron microscopy. We found a drop in the amplitude of the cochlear microphonics and compound action potentials 2 to 2.5 hours after injection. There were also changes in the morphology of stereocilia and the tectorial membrane. The stereocilia lost their rigidity and the tectorial membrane appeared swollen. These effects were less severe in animals which were pretreated with dexamethasone.

Action Potentials↗

PubMed1991

Preparation of inner ear sensory hair bundles for high resolution scanning electron microscopy.

Chemical fixation techniques for preservation of sensory hair bundles in the mammalian inner ear for scanning electron microscopy (SEM) are reviewed. Fixatives employed were glutaraldehyde, glutaraldehyde-picrate, glutaraldehyde-tannic acid, glutaraldehyde-formaldehyde, glutaraldehyde followed by postfixation with osmium tetroxide and the osmium thiocarbohydrazide (OTOTO) method. Dehydration was routinely accomplished with ascending grades of acetone followed by critical point drying with liquid CO2 or fluorocarbon sublimation. Specimens other than those prepared by the OTOTO method were metal coated with gold, gold-palladium or platinum. Material was viewed at high resolution (2-3 nm) in a transmission electron microscope (TEM) fitted with a scanning system and an LaB6 filament. A few specimens, which were either coated with platinum, carbon or uncoated, were examined in a field emission SEM. We have concluded that glutaraldehyde fixation followed by critical point drying with CO2 and coating with platinum gives the best general preservation of stereocilia and their cross-links for routine high resolution SEM, but that carbon-coated or uncoated specimens offer potentially better results free from metal coating artifacts when viewed with field emission SEM. These methods have enabled us to make novel observations upon the surface detail and cross-links of stereocilia which have helped considerably in understanding the mechanical properties of hair bundles particularly in relation to sensory transduction. We have found that stereocilial surface detail and cross-links are sensitive to fixation regimens. In particular they are degraded by exposure to osmium tetroxide; they are also highly labile since deleterious changes in their appearance can be detected as early as 15 minutes following death.

Animals↗

PubMed1990

High resolution scanning electron microscopy of stereocilia in the cochlea of normal, postmortem, and drug-treated guinea pigs.

The morphology of hair bundles has been studied by high resolution scanning electron microscopy using a variety of fixatives, including glutaraldehyde, glutaraldehyde-picrate, glutaraldehyde-tannic acid, glutaraldehyde followed by post-fixation in osmium tetroxide, and the osmium thiocarbohydrazide technique. Critical evaluation of several metal coatings, gold, gold-palladium, and platinum has been carried out. Both the surface texture of stereocilia and their cross-links are sensitive to fixation and metal coating. We are of the opinion that glutaraldehyde gives the best general quality of fixation and preservation for all types of cross-links. We have described three major sets of cross-links: first, lateral links connecting stereocilia within the same row; second, lateral links connecting stereocilia of adjacent rows; and third, upward-pointing links, one per stereocilium, connecting the tip of each shorter stereocilium to the lateral surface of the adjacent taller stereocilium. Current physiological and anatomical evidence suggests that the lateral links couple the individual stereocilia within the hair bundle so that they function as a single mechanical unit. The upward-pointing tip links are ideally placed to respond to mechanical deformation of the hair bundle, being stretched when the stereocilia are deflected in the excitatory direction towards the tallest row and relaxed when deflected in the opposite, inhibitory direction. Postmortem morphological changes are detected within 15 minutes of cardiac arrest and become progressively more pronounced in time. These results enabled us to distinguish specific drug-induced changes which could not be attributed simply to cell death. Effects of cisplatin and kanamycin upon hair bundles are described. The work reported here is based on studies using the guinea pig cochlea. Some of the postmortem changes described have also been confirmed in human cochleas. It is stressed that many of the postmortem and drug-induced effects can only reliably be studied by high resolution scanning electron microscopy coupled with appropriate preparation procedures.

Animals↗

PubMed1989

Tip-link organization in anomalously-oriented hair cells of the guinea pig cochlea.

Stereocilia are described in a group of guinea pigs with abnormal conformation of the outer hair cell stereociliary bundles. Rows of stereocilia were found which were circular instead of V- or W-shaped, and rotated or even reversed in orientation with respect to normal. The stereocilia have however a normal gradation in heights of the rows of stereocilia, and have tip links running in the direction of gradation in height.

Animals↗

PubMed1989

The organization of tip links and stereocilia on hair cells of bird and lizard basilar papillae.

Auditory papillae from three species of bird (pigeon, starling, and chick), and two species of European lizard (Podarcis muralis and Podarcis sicula) were examined by scanning electron microscopy. Hair bundles from all papillae showed tip links oriented along the direction of gradation in heights of the stereocilia (i.e. parallel to the hair-cell axis of bilateral symmetry, and so parallel to the excitatory-inhibitory axis for mechanotransduction). This orientation was seen irrespective of the overall orientation of the hair bundle within the papilla. The stereocilia formed columns, joined by the tip links, which ran parallel to the hair-cell axis of bilateral symmetry. The stereocilia within the same column tended to stay together, while those in different columns tended to separate during preparation. In many columns all the stereocilia tended to be a little taller, or a little shorter, than the equivalent stereocilia in adjacent columns, suggesting that all the stereocilia within one column had been affected by a common height determinant during development. In addition, links running laterally between stereocilia were seen, in a band near the base of the stereocilia. The results are consistent with the hypothesis that tip links are a universal feature of mechano-transducing acousticolateral hair cells, and that they are involved in sensory transduction. The results also support suggestions that the tip links may play a role in determining the heights of the stereocilia during development.

Animals↗

PubMed1988

Further observations on the fine structure of tip links between stereocilia of the guinea pig cochlea.

Stereocilia of the guinea pig organ of Corti were examined by transmission electron microscopy, after fixation in glutaraldehyde and tannic acid, and postfixation and en bloc staining in osmium tetroxide, tannic acid, uranyl acetate, and phosphotungstic acid. Tip links were observed between the stereocilia. The links emerged from the tips of the shorter stereocilia in the hair bundle, running nearly at right angles to the cuticular plate, to join the side-wall of the adjacent taller stereocilium of the next row. The tip links had a fine filamentous core, approximately 6 nm in diameter. The core was surrounded by positively-staining amorphous material, which had a variable appearance from link to link. The central filament inserted into membrane specialisations at both its upper and lower ends. The results suggest that tip links have two components, and that the central filament, which has the same diameter as an actin filament, is suitable for transmitting stimulus-induced movements to the transducer channels of the stereocilium. The central filament would therefore concentrate the stimulus-induced forces onto a small area of cell membrane.

Animals↗

PubMed1987

Vulnerability of tip links between stereocilia to acoustic trauma in the guinea pig.

The cochleae of anaesthetized guinea pigs were prepared for scanning electron microscopy, immediately after exposure to an intense tone. Stereocilia on hair cells showing relatively small degrees of disruption were analyzed. If the bundles of stereocilia showed no or only a very slight degree of disorganization, the fine links emerging from the tips of the shorter stereocilia remained intact. If the stereocilia were separated more than a very little, the tip links between stereocilia were no longer visible. However, it was possible for tip links to remain intact in some parts of the hair bundle, while tip links in other, more disrupted parts, were lost. In outer hair cells, tip links did not seem any more vulnerable in one position than in another. In inner hair cells, it was commonly found that the tip links running between the tallest stereocilia and the next row of shorter stereocilia had broken, while the tip links running between the other shorter rows of stereocilia remained intact. The results suggest that tip links between stereocilia are preserved as long as the other links between the stereocilia and the cytoskeleton of the stereocilium remain intact. When the latter are damaged the tip links fracture. The results also suggest that, if the tip links are indeed involved in transduction, some degree of stimulus transduction can continue in damaged inner hair cells, albeit with a reduced sensitivity.

Acoustic Stimulation↗

PubMed1987

The effect of chronic application of kanamycin on stereocilia and their tip links in hair cells of the guinea pig cochlea.

Albino guinea pigs were treated with kanamycin (400 mg/kg i.p.) daily for 10 days. After a 2-week recovery period their cochleae were analyzed by scanning electron microscopy. Attention was paid to those outer hair cells which had been less severely damaged. Stereocilia of the outer hair cells were often constricted at the root, and in some cases had become detached at the root. In stereocilia showing any degree of abnormality, the tip links were usually missing. However, in a few exceptional cases, tip links could remain on stereocilia showing other abnormalities, such as constriction at the root. Where hair cells were otherwise apparently unaffected, a much higher proportion of tip links remained, even on cells situated in an area of extensive hair cell loss. The results give further information on the process of kanamycin poisoning. They also suggest that substantial loss of tip links, and therefore perhaps of transduction, is one of the preliminary consequences of kanamycin poisoning.

Animals↗