Biomedical subjects

D F Dolan

Publications and source records attributed to D F Dolan.

At least 19 recordsLinked to original sources

PubMed2004

Skeletal dysplasia and male infertility locus on mouse chromosome 9.

In mice and humans, growth insufficiency and male infertility are common disorders that are genetically and phenotypically complex. We describe a spontaneously arising mouse mutant, chagun, that is affected by both dwarfism and male infertility. Dwarfism disproportionately affects long bones and is characterized by a defect in the proliferative zone of chondrocytes in the growth plate. Gonads of mutant males are small, with apparent germ cell loss and no evidence of mature sperm. The locus responsible for chagun is recessive and maps to distal chromosome 9, in a region homologous to human chromosome 3. This location is consistent with chagun defining a novel locus. Identification of the mutant gene will uncover the basis for another type of skeletal dysplasia and male infertility.

Animals↗

PubMed2003

Frequency-specific effects on cochlear responses during activation of the inferior colliculus in the Guinea pig.

The inferior colliculus (IC) is a major processing center in the ascending auditory pathway. The role of the IC in the descending efferent auditory system is less clear. Although the IC central nucleus (ICC) is the major relay station for the ascending auditory pathways, the IC's cortex receives its main input from the neocortex and nonauditory sources. The goal of this study was to determine if the IC subdivisions had different functions in the descending efferent auditory system. IC subdivisions were identified by their tuning curves evoked by tone stimulation, and the effects of localized electrical stimulation on the cochlear whole-nerve action potential (CAP). Sharp tuning curves were obtained from ICC in contrast to broad tuning curves from the lateral, external cortex. Electrical stimulation within the central nucleus had a sharply tuned effect on the CAP. The frequency region affected within the cochlea closely matched the best frequency of local cells within the central nucleus. The effect of electrical stimulation within the lateral, external cortex on the CAP was smaller in comparison to central nucleus stimulation. Similar to the broad tuning of cells within the lateral cortex, electrical stimulation had a broad frequency effect on CAP thresholds.

Action Potentials↗

PubMed2001

Severe vestibular and auditory impairment in three alleles of Ames waltzer (av) mice.

The genetic and physiological characterization of circling, hearing-impaired mouse mutants has greatly facilitated our understanding of non-syndromic sensorineural deafness, the most common form of hereditary human hearing loss. Here we report the first phenotypic characterization of three alleles of Ames waltzer (av). Neither electrical potentials (auditory brainstem response) nor behavioral responses to sound could be evoked in any of the three alleles at any age or frequency. However, the endocochlear potential was found to be normal, indicating that the primary pathology is not in the stria vascularis. To determine the earliest changes and help identify the primary causes of deafness in av, we performed morphological studies in 15-16 day old mutants, just prior to the maturation of the cochlea. Although av(2J) is slightly more affected than the other two alleles, our studies show a high similarity between all three alleles. The first detectable changes are observed in the stereocilia and cytoplasm of hair cells, and in the cellular shape and microvilli of supporting cells. These changes are followed by degeneration of the cochlear and vestibular neuroepithelium.

Alleles↗

PubMed2000

Glial cell line-derived neurotrophic factor has a dose dependent influence on noise-induced hearing loss in the guinea pig cochlea.

We examined the effectiveness of glial cell line-derived neurotrophic factor (GDNF) to attenuate cochlear damage from intense noise stress. Subjects were exposed to 115 dB SPL one octave band noise centered at 4 kHz for 5 h. They received artificial perilymph with or without GDNF into the left scala tympani at 0.5 microliter/h from 4 days before noise exposure through 8 days following noise exposure. Different concentrations of GDNF (1 ng/ml, 10 ng/ml, 100 ng/ml, and 1 microgram/ml) were applied chronically directly into the guinea pig cochlea via a microcannula and osmotic pump. Noise-induced hearing loss was assessed with pure tone auditory brainstem responses (at 2, 4, 8 and 20 kHz), measured prior to surgery, 1 day before noise exposure, and 7 days following noise exposure. Subjects were killed on day 8 following exposure for histological preparation and quantitative assessment of hair cell (HC) damage. A dose-dependent protective effect of GDNF on both sensory cell preservation and hearing function was found in the treated ears. At 1 ng/ml, GDNF showed no significant protection; at 10 ng/ml, GDNF showed significant HC protection; and at 100ng/ml, it was greater and bilateral. At 1 microgram/ml, GDNF appeared to have a toxic effect under noise stress in some cochleae. These findings indicate that GDNF at certain concentrations can effectively protect the inner ear from noise-induced hearing loss.

Animals↗

PubMed2000

The effects of efferent activation on the acoustically and electrically evoked otoacoustic emission.

The effects of efferent activation on the otoacoustic emission were measured in anesthetized guinea pigs. The otoacoustic emission (2F(1)-F(2)) was evoked by the conventional method of presenting either two continuous tones or a sinusoidal current to the round window (RW) of the cochlea. The efferent effects on the acoustically evoked emission are greatest at low stimulus levels and least for high levels. The efferent effects on the electrically evoked emission (EEOAE) are relatively constant across current levels. In each case, efferent activation resulted in an initial large reduction in the emission amplitude followed by a smaller and more constant reduction. Strychnine eliminated the efferent effects independent of the method of emission activation. Strychnine had no effect on the EEOAE, suggesting that the RW current did not evoke a local efferent effect. Slow versus fast efferent effects were observed in the recovery of the emission amplitude at the termination of efferent activation. Only a fast recovery in the emission amplitude was observed for stimuli below 10 kHz while the amplitude recovery had fast and slow components for stimuli presented above 10 kHz.

Acoustic Stimulation↗

PubMed2000

Hair cells in the inner ear of the pirouette and shaker 2 mutant mice.

The shaker 2 (sh2) and pirouette (pi) mouse mutants display severe inner ear dysfunction that involves both auditory and vestibular manifestation. Pathology of the stereocilia of hair cells has been found in both mutants. This study was designed to further our knowledge of the pathological characteristics of the inner ear sensory epithelia in both the sh2 and pi strains. Measurements of auditory brainstem responses indicated that both mutants were profoundly deaf. The morphological assays were specifically designed to characterize a pathological actin bundle that is found in both the inner hair cells and the vestibular hair cells in all five vestibular organs in these two mutants. Using light microscope analysis of phalloidin-stained specimens, these actin bundles could first be detected on postnatal day 3. As the cochleae matured, each inner hair cell and type I vestibular hair cell contained a bundle that spans from the region of the cuticular plate to the basal end of the cell, then extends along with cytoplasm and membrane, towards the basement membrane. Abnormal contact with the basement membrane was found in vestibular hair cells. Based on the shape of the cellular extension and the actin bundle that supports it, we propose to name these extensions "cytocauds." The data suggest that the cytocauds in type I vestibular hair cells and inner hair cells are associated with a failure to differentiate and detach from the basement membrane.

Actin Cytoskeleton↗

PubMed1999

Acquired resistance to acoustic trauma by sound conditioning is primarily mediated by changes restricted to the cochlea, not by systemic responses.

Hearing loss caused by intense sound exposure can be significantly reduced by pre-exposing subjects to moderate-level acoustic stimuli. This phenomenon occurs in a variety of mammals. We investigated whether sound conditioning provides acquired resistance to acoustic trauma through local mechanisms selectively in the conditioned ears or if systemic mechanisms are involved that would yield contralateral protection in unconditioned ears. Guinea pigs (group I) in which one external ear canal was occluded were exposed to conditioning sound (2-20 kHz, 85 dB SPL, 5 h/day, 10 days). After removing the occlusion, the animals were then subjected bilaterally to intense noise (2-20 kHz, 110 dB SPL, 5 h) 5 days after the last conditioning exposure. Animals without ear canal occlusion were also exposed to the intense sound without conditioning (group II) or following the same conditioning exposure (group III). Intense sound exposure caused significantly greater permanent ABR threshold shifts at all frequencies tested (4, 8, 12, 16 and 20 kHz) in group II than in group III. In group I, the occluded ears showed significantly greater threshold shifts at all frequencies compared to the unoccluded ears. The threshold shifts in the occluded ears in group I were identical to those observed in group II; and the shifts in unoccluded ears in group I were identical to those in group III. Protective effects provided by sound conditioning were almost the same in group III and in the unoccluded ears in group I. The extent of hair cell damage supported the physiological findings. These results indicate that acquired resistance to acoustic trauma provided by sound conditioning is restricted to the cochlea exposed to conditioning sound, suggesting that conditioning protection is mediated primarily by the changes that occur locally within the conditioned cochlea. This animal model, with unilateral external ear canal occlusion during sound conditioning, is useful for studies of the mechanisms of conditioning protection.

Acoustic Stimulation↗

PubMed1999

KHRI-3 monoclonal antibody-induced damage to the inner ear: antibody staining of nascent scars.

Intracochlear infusion of the KHRI-3 monoclonal antibody results in in vivo binding to guinea pig inner ear supporting cells, loss of hair cells and hearing loss. To further characterize the basis for KHRI-3-induced hearing loss, antibody was produced in a bioreactor in serum-free medium, affinity purified, and compared to conventionally prepared antibody by infusion into the scala tympani using mini-osmotic pumps. In vivo antibody binding was observed in 10 of 11 guinea pigs. A previously unreported pattern of KHRI-3 antibody binding to cells involved in scar formation was noted in five guinea pigs. All but one of the KHRI-3-infused animals demonstrated a hearing loss of > 10 dB in the treated ear. In five of 11 animals the threshold shift was 30 dB or more, and all had hair cell losses. In one guinea pig infused with 2 mg/ml of antibody, the organ of Corti was absent in the basal turn of the infused ear. This ear had a 45-50 dB threshold shift but, curiously, no detectable antibody binding in the residual organ of Corti. Organ of Corti tissue was fragile in antibody-infused ears. Breaks within the outer hair cell region occurred in 5/11 infused ears. The contralateral ears were normal except for one noise-exposed animal that demonstrated hair cell loss in the uninfused ear. Three animals were exposed to 6 kHz noise (108 dB) for 30 min on day 7. Antibody access to the organ of Corti may be increased in animals exposed to noise, since the strongest in vivo binding was observed in noise-exposed animals. Loss of integrity of the organ of Corti seems to be the primary mechanism of inner ear damage by KHRI-3 antibody. The binding of KHRI-3 antibody in new scars suggests a role of the antigen in scar formation. Antibodies with binding properties similar to KHRI-3 have been detected in 51% of patients diagnosed with autoimmune sensorineural hearing loss; thus, it seems likely that such autoantibodies also may have pathologic effects resulting in hearing loss in humans.

Animals↗

PubMed1999

Morphological and physiological effects of long duration infusion of strychnine into the organ of Corti.

Acute strychnine administration has long been used as a method to eliminate the effects of efferent activity. It has been shown that long after termination of chronic strychnine infusion into the cochlea, the ear becomes more susceptible to acoustic trauma suggesting that chronic strychnine infusion results in long lasting or permanent disruption of efferent function. Much research has been directed towards the functional significance of the olivocochlear system. However, there is little information concerning the effect of long duration inactivation of the medial olivocochlear system in an awake behaving animal. This study was designed to determine the structural and functional consequences of inactivation of the efferents by chronic infusion of strychnine into the cochlear perilymph of guinea pigs for two weeks via an osmotic pump. Physiological evaluations showed that the strychnine infusion eliminated the efferent induced reduction of the cochlear whole-nerve action potential three weeks after cessation of strychnine infusion. Contralateral efferent function remained unaltered. Histological evaluation at the light and electron microscopic levels revealed disoriented efferent synapses under the outer hair cells.

Acoustic Stimulation↗

PubMed1998

The medial cochlear efferent system does not appear to contribute to the development of acquired resistance to acoustic trauma.

Noise-induced hearing loss (NIHL) was compared between sound conditioned and unconditioned guinea pigs, in which the left ear in both groups had been perfused with strychnine. Animals in the conditioned group were subjected to moderate sound (85 dB SPL broadband, 5 h/day, 10 days) and then exposed to intense sound (110 dB SPL broadband, 5 h). Unconditioned animals were exposed only to the intense sound. Following intense sound exposure, strychnine-treated ears showed greater NIHL than untreated ears in both unconditioned and conditioned animals, demonstrating the role of the medial efferents to reduce NIHL. Conditioned animals, however, showed smaller hearing loss and cochlear damage in both strychnine-treated and untreated ears compared to unconditioned animals; the protective effects given by conditioning were equivalent between the strychnine-treated and untreated ears. These results suggest that, although the medial efferent system acts to attenuate NIHL, it may not be necessary for the acquired resistance to NIHL provided by conditioning.

Acoustic Stimulation↗

PubMed1998

Genetic mapping refines DFNB3 to 17p11.2, suggests multiple alleles of DFNB3, and supports homology to the mouse model shaker-2.

The nonsyndromic congenital recessive deafness gene, DFNB3, first identified in Bengkala, Bali, was mapped to a approximately 12-cM interval on chromosome 17. New short tandem repeats (STRs) and additional DNA samples were used to identify recombinants that constrain the DFNB3 interval to less, similar6 cM on 17p11.2. Affected individuals from Bengkala and affected members of a family with hereditary deafness who were from Bila, a village neighboring Bengkala, were homozygous for the same alleles for six adjacent STRs in the DFNB3 region and were heterozygous for other distal markers, thus limiting DFNB3 to an approximately 3-cM interval. Nonsyndromic deafness segregating in two unrelated consanguineous Indian families, M21 and I-1924, were also linked to the DFNB3 region. Haplotype analysis indicates that the DFNB3 mutations in the three pedigrees most likely arose independently and suggests that DFNB3 makes a significant contribution to hereditary deafness worldwide. On the basis of conserved synteny, mouse deafness mutations shaker-2 (sh2) and sh2J are proposed as models of DFNB3. Genetic mapping has refined sh2 to a 0.6-cM interval of chromosome 11. Three homologous genes map within the sh2 and DFNB3 intervals, suggesting that sh2 is the homologue of DFNB3.

Alleles↗

PubMed1998

Interactive effects of aging with noise induced hearing loss.

Aging is associated with a down-regulation in metabolism, which may underlie an increased sensitivity to stress agents and a decreased repair of tissues following stress. In the auditory system this could lead to increased sensitivity to noise induced hearing loss (NIHL) with age. This study examines the effects of high intensity noise exposure (0.5-40 kHz noise at 108 dB SPL for 45 min) on young and old normal (CBA/Ca) mice and young premature presbycusis (C57BL/6) mice. Tone evoked auditory brainstem response (ABR) audiograms were obtained before and after the exposure. The animals were sacrificed and cytocochleograms were performed. The ABR threshold shifts and hair cell losses which followed noise exposure increased with increasing age in the normal (CBA/Ca) mice. Subjects which showed early presbycusis (C57BL/6) associated with vascular pathology, showed an increased sensitivity to noise induced hearing loss over normal subjects. Interestingly, in some cases the physiological loss in C57BL/6 mice was not associated with hair cell loss. These findings support the view that aging with or without hearing loss increased the sensitivity of the ear to NIHL. However, it seems that other factors may contribute to NIHL in the animals with premature presbycusis.

Aging↗

PubMed1997

Chronic strychnine administration into the cochlea potentiates permanent threshold shift following noise exposure.

To investigate whether elimination of the medial efferent system influences permanent threshold shift following noise exposure, we developed an animal model in which strychnine was chronically delivered into the cochlea via an osmotic pump. Pigmented female guinea pigs were allocated into three groups: group I was treated with strychnine (50 microM, 0.5 microl/h, 14 days) in the left ear and exposed to noise (105 dB SPL broadband, 3 h) 3 weeks after the cessation of the strychnine perfusion; group II received strychnine in the left ear but no noise exposure; group III was treated with Ringer's solution in the left ear and exposed to noise. Animals in group II developed no hearing loss after the strychnine perfusion. The operated ears in group I demonstrated greatest hearing threshold shift 3 h after noise exposure. Hearing recovered during 2 weeks after noise exposure in both operated and non-operated ears in groups I and III. Two weeks after noise exposure, the operated ears in group I showed significantly greater threshold shift at 12, 16, and 20 kHz compared to the operated ears in group III and non-operated ears in groups I and III. These findings suggest that chronic strychnine administration into the cochlea inactivates the medial efferents without changing hearing threshold and that the medial efferents help to protect against permanent threshold shift following noise exposure.

Animals↗

PubMed1997

Basilar membrane velocity noise.

Basilar membrane (BM) noise, measured as a velocity signal under the quiet acoustic condition, was investigated in the guinea pig. The cochleas of anesthetized young healthy guinea pigs were surgically exposed and a hole was made on the lateral wall of the scala tympani of the first cochlear turn for visualization of the BM and measurement of the BM velocity with a laser interferometer. The amplitude and frequency of the BM velocity noise were analyzed by a spectrum analyzer under different conditions. The spectrum of the BM velocity noise was a band limited function with a peak velocity at the topographic best frequency of the measured location on the BM. The peak velocity ranged to about 8 microm/s and depended on the physiological condition of the cochlea. Saline blockage of the external auditory canal or the middle ear did not change the BM noise. BM noise was much smaller, or was not evident, when the cochlear sensitivity decreased. The suppression tuning curve of the BM velocity noise indicates that the maximum suppression caused by an acoustic pure tone occurred at the best frequency location. A low sound level wide band acoustic noise given to the external ear canal produced a spectrum function having the same frequency and amplitude response as the BM noise. Electrical stimulation of the crossed olivocochlear bundle significantly depresses the BM velocity noise. These data demonstrate that the BM noise is a representation of internal rather than external noise. The amplitude and frequency of the BM noise reflect the usual cochlear sensitivity and frequency selectivity. Since the organ of Corti in the sensitive cochlea is a highly sensitive and tuned mechanical system, the internal (to the animal) noise responsible for the BM noise may originate from mechanical vibrations remote from the cochlea and propagated to the ear, or may be caused by Brownian motion of cellular structures in the cochlea.

Acoustic Stimulation↗

PubMed1997

Frequency-dependent enhancement of basilar membrane velocity during olivocochlear bundle stimulation.

Basilar membrane (BM) velocity responses were measured in the presence of olivocochlear bundle (OCB) stimulation. Frequency threshold tuning curves (FTCs) were derived from tone-evoked input-output (I/O) functions. Efferent nerve activation produced decreases in velocity amplitude for frequencies around best frequency (BF) at low stimulus levels with little or no effect for stimuli well below the BF. A level-dependent efferent reduction/enhancement of BM velocity was found for certain stimulus frequencies above the BF. Efferent activation either had no effect or caused small reductions in the velocity response produced by low level sound, whereas, at higher stimulus levels, efferent activation increased the velocity response. The derived FTCs, therefore, showed criterion-dependent changes with efferent activation. For low BM criterion velocities, FTCs showed the classic desensitization of the tip region without a shift of BF. Some BM velocity criterion values showed FTCs with an expanded high-frequency response area, also without a shift of BF. The results suggest that the effect of OCB activation changes the gain of the voltage-dependent outer hair cell motility such that BM velocity response near BF is decreased while increasing the response for tones well above BF.

Acoustic Stimulation↗

PubMed1997

The mechanism and site of action of lidocaine hydrochloride in guinea pig inner ear.

Lidocaine was applied to the round window (RW) in order to localize its site of action in the cochlea. Cochlear microphonic (CM), summating potential (SP), and compound action potential (CAP) input/output functions were measured to a 16 kHz tone burst to assess the functional changes of the cochlea. In separate experiments, the effect of lidocaine on the whole cell current of isolated outer hair cells (OHC) was studied. A dose of 2 microliters of 40 mM lidocaine in saline solution, when applied to the RW, caused a small change in all measured variables, indicating a passage of the drug through the RW membrane to sites of action. However, 160 mM of lidocaine further decreased CM, SP, and CAP by a total of 40% from the control. A partial recovery occurred for CM during the 30 min follow-up period. CAP and SP continued to decline. In isolated OHCs, lidocaine decreased the whole cell current in a dose-dependent fashion. The KD for lidocaine effect on OHCs was 7 mM. Our in vivo results indicate that lidocaine affects OHCs and reduces CM, causing a subsequent reduction in SP and CAP. The increased effect of lidocaine on CAP and SP, while CM is recovering, suggests an additional specific effect of lidocaine on the cochlear nerve and/or on inner hair cells. Considering that lidocaine alters OHC current (in isolated hair cells) and that lidocaine does not affect endocochlear potential [Laurikainen et al. Acta Otolaryngol (Stockh) 1991: 112: 800-9], the observed CM changes are most likely due to an in vivo effect on OHCs. Thus, the early effect of lidocaine on the cochlea appears to be due to a significant change in organ of Corti function, rather than to direct anesthesia of the cochlear nerve. Later, an independent effect of the drug may occur on neural tissues in the inner ear.

Animals↗

PubMed1996

Age-related differences in cochlear microcirculation and auditory brain stem response.

OBJECTIVE: To examine possible age-related differences in auditory sensitivity and cochlear vascular properties. DESIGN: This study is designed to provide information regarding cochlear function using physiological and audiological measures. Each animal underwent intravital microscopic evaluation of red blood cell velocity, vessel diameter, and vascular permeability in the second turn of the cochlear lateral wall. Auditory brain stem responses were used to determine hearing sensitivity. SUBJECTS: Four age ranges of male Fischer rats were studied: young, 2 to 4 months (n = 9); mid-young, 9 to 11 months (n = 8); mid-old, 18 to 20 months (n = 6); and old, 30 to 34 months (n = 10). RESULTS: Auditory brain stem response testing showed an age-related decrease in auditory sensitivity. Intravital microscopic analysis showed age-related statistically significant decreases in red blood cell velocity and increased vascular permeability with a trend for reduced capillary diameters. CONCLUSIONS: The process of aging is associated with many biochemical and physiological changes that include decrease in cellular water concentration, ionic changes, and decreased elasticity of cellular membranes. One contributing factor to this process may be altered vascular characteristics, such as reduced flow and vascular plasticity, as well as increased vascular permeability. These age-related changes may result in reductions in oxygen and nutrient delivery, and also waste elimination. Our results suggest that progressive age-associated vascular compromise may be a contributing factor in presbycusis.

Age Factors↗

PubMed1996

Steady-state sinusoidal velocity responses of the basilar membrane in guinea pig.

This report provides a detailed analysis of tone-evoked velocity responses of basilar membrane (BM) motion measured in the basal turn of the guinea pig cochlea. A laser Doppler vibrometer, coupled to a compound microscope, measured the velocity of reflective microbeads placed onto the scala tympani surface of the BM. The velocity responses of the stapes footplate were also determined, allowing the calculation of a BM transfer function. The normal transfer function is compared to that seen with cochlear "insensitivity" and postmortem. This comparison results in measures of the active process contribution to the magnitude and phase of the BM transfer function. It was found that the active process contributed as much as 65 dB of "gain" and 270 degrees of phase lag at the best frequency. Other details about features of input/output velocity functions and the derived mechanical frequency-tuning curve at a criterion of 50 micron/s are analyzed.

Acoustics↗