PubMed Health⌕ Search

Biomedical subjects

N K Woolf

Publications and source records attributed to N K Woolf.

At least 19 recordsLinked to original sources
PubMed1996

Improved resolution of fibronectin mRNA expression in the inner ear using laser scanning confocal microscopy.

We describe a modified in situ hybridization protocol for localizing and quantifying fibronectin gene expression at the cellular level in paraffin sections of rat temporal bone. When combined with a novel analytical approach using laser scanning confocal microscopy (LSCM), this protocol significantly improved the resolution, sensitivity, and specificity of existing procedures for evaluating fibronectin synthesis in developing inner ear. For simultaneous viewing of cochlear anatomy and the autoradiographic signal, transmitted light images of the cochlea were collected separately from LSCM reflected light images of the autoradiographic silver grains and then the two images were electronically merged. Within the first 2 microns below the surface of the emulsion, silver grains were clustered specifically over hybridized cells. In contrast, nonspecific silver grain development (i.e., background noise) was confined primarily to the lower 5 microns of the emulsion adjacent to the tissue section. Limiting the volume of the emulsion examined in the LSCM analysis, i.e., restricting the range of optical sectioning to the first 2 micron below the surface of the emulsion, effectively minimized nonspecific background noise and maximized the specificity of the hybridization signal. The improvements offered by the described methodological approaches are equally appropriate for non-calcified tissues.

Animals↗
PubMed1994

Protection from noise-induced hearing loss by prior exposure to a nontraumatic stimulus: role of the middle ear muscles.

Recent evidence suggests that prior exposure to a moderate-level acoustic stimulus can reduce damage due to later exposure to the same stimulus at high intensity [Canlon et al., Hear. Res. 34, 197-200 (1988)]. To test the role of the middle ear muscles (MEMs) in this phenomenon, Mongolian gerbils were conditioned by exposure to a two-octave band of noise (1414-5656 Hz) at 81 dB SPL for 3 weeks. Either immediately afterward, or following a one week rest period, they were exposed to the same stimulus at 110 dB SPL for one hour. The ABR thresholds of these animals were compared to those seen in animals exposed at 110 dB SPL without conditioning. The MEMs of one ear in each subject were cut, to determine their role in any noise trauma protection effects. In the unoperated ears, conditioning without a recovery period did not alter the effects of the 110 dB stimulus. Conditioning followed by a one week recovery period reduced both temporary (TTS) and permanent (PTS) threshold shift. MEM section had no effect on either TTS or PTS in unconditioned subjects, and did not alter the reduction in TTS or PTS seen with conditioning. It is concluded that the noise trauma resistance provided by acoustic conditioning is not mediated by the MEMs.

Acoustic Stimulation↗
PubMed1993

Fibronectin-like immunoreactivity of the basilar membrane of young and aged rats.

Dysfunction of cochlear mechanics has been hypothesized to be a source of age-related hearing loss and the basilar membrane mass and stiffness contribute to normal cochlear mechanics. Fibronectin, a large, extracellular matrix protein and a major component of the basilar membrane, may contribute to both the mass and stiffness of the membrane. Mesothelial cells underlying the basilar membrane may produce the fibronectin and also contribute to the mass of the membrane. Changes in either the fibronectin or the mesothelial cells might, therefore, have an effect on cochlear mechanics. In order to assess basilar membrane changes in aged animals, young adult (2-4 months) and aged (24-26 months) Sprague-Dawley rats were evaluated for the presence of fibronectin-like protein and mesothelial cells. The basilar membrane in the young animals had strong fibronectin-like immunoreactivity throughout its length. The old animals, on the other hand, showed normal fibronectin immunoreactivity in the basilar membrane of the basal turn, but little or no reactivity in the apical cochlear turn. The number of mesothelial cells was reduced throughout the length of the membrane in aged animals, with the greatest loss in the basal turn (60% fewer cells). These two degenerative changes, which appear to be independent of each other, may contribute to the observed threshold shifts in aged cochleas.

Aging↗
PubMed1992

Immunohistochemical localization of fibronectin-like protein in the inner ear of the developing gerbil and rat.

Immunohistochemistry was used to demonstrate the distribution of fibronectin-like protein within the developing inner ear of two species of altricial rodents: gerbils and rats. While there were temporal differences between the two species, the developmental sequence of immunostaining was virtually identical. Most notably, in rats from embryonic day 18 through day 1 postpartum, and in gerbils from birth through day 4 postpartum, intense, discrete fibronectin-like immunoreactivity was observed in the cochlea immediately beneath the inner and outer hair cells, sites of active auditory nerve fiber growth and nerve-hair cell synaptogenesis at these ages. The results suggest that fibronectin is appropriately positioned spatially and temporally to play a significant role in promoting, guiding and/or maintaining neural innervation within the developing organ of Corti. The temporo-spatial pattern of immunostaining in Schwann cells and auditory (VIIIth cranial) nerve neurons implies that fibronectin also plays a significant role in the early formation of myelin. In non-neural elements of the cochlea, fibronectin is a major structural component within the basilar membrane at all of the developmental stages investigated.

Animals↗
PubMed1992

Central auditory metabolic activity induced by intense noise exposure.

Neural activity in the central auditory system was mapped by measuring 2-deoxyglucose (2-DG) uptake during a one hour exposure to a two-octave (1414-5656 Hz) band of noise. Gerbils were exposed to 100, 110 or 120 dB SPL, intensities which can produce only temporary (100 dB) or both temporary and permanent (120 dB) hearing loss. Exposure to 100 dB SPL evoked high levels of neural activity throughout responsive regions of auditory nuclei. At 110 dB SPL, a central region of low neural activity was surrounded by areas exhibiting increased activity. At 120 dB SPL, neural activity was low in almost all areas of auditory nuclei. To study the effects of permanent hearing loss on auditory neuronal activity, other animals were given 2-DG during exposure to 65 dB SPL broad band noise as a test stimulus, two months after exposure to the noise band at 110 dB SPL. Central auditory nuclei showed a tonotopic region of low neural activity corresponding to an approximately 3 kHz pure tone, surrounded by regions of evoked activity. The deficits in evoked metabolic activity observed both during and long after noise exposure appear to exceed those predicted from the degree of temporary and permanent threshold shift produced by the same noise exposures.

Acoustic Stimulation↗
PubMed1990

Spatial distribution of neural activity evoked by electrical stimulation of the cochlea.

Activity in the central auditory system was mapped with 2-deoxyglucose (2-DG) autoradiography, using either pure tones or electrical stimulation of the normal cochlea. Electrical stimulation with both monopolar (distant reference electrode) and bipolar prostheses near threshold increased 2-DG uptake in auditory nuclei in a manner similar to that seen with a pure tone: increased 2-DG uptake was restricted to a small frequency region of brainstem and mid-brain auditory nuclei. The position of this area was related to the cochlear location of the prosthesis. At higher current amplitudes only the bipolar prosthesis retained spatial restriction of evoked neural activity, while stimulation through a monopolar prosthesis produced evoked activity in all frequency regions of auditory nuclei, and in non-auditory nuclei. Activation of non-auditory structures was consistent with spread of current through the brainstem, rather than activation of peripheral nerves. At all current amplitudes, a monopolar prosthesis evoked higher levels of 2-DG uptake than a bipolar prosthesis. The results suggest that while a bipolar prosthesis provides greater spatial restriction of evoked neural activity and a greater dynamic range, a monopolar prosthesis produces higher levels of evoked activity.

Acoustic Stimulation↗
PubMed1989

Auditory neural activity evoked by pure-tone stimulation as a function of intensity.

The 2-deoxyglucose (2-DG) autoradiographic technique was employed to map activation of the central auditory pathway in the mongolian gerbil during stimulation with a 3.0 kHz tone at several intensities. In most auditory nuclei, the tone produced restricted areas in which 2-DG uptake was markedly higher than that of adjacent tissue, at locations consistent with the known tonotopic organization of the structure. The size of these regions changed relatively little with increasing stimulus intensity from 25 to 65 dB SPL (re 0.0002 dyne/cm2). At higher intensities, evoked uptake spreads into locations which represent frequencies above 3.0 kHz. In the inferior colliculus, relative 2-DG uptake decreased with increasing stimulus intensity in bands on either side of the 3.0 kHz region. These bands of reduced uptake became wider with increasing stimulus intensity from 25 to 85 dB SPL. The optical densities of auditory structures were normalized by the density of non-auditory white matter to derive optical density ratios. In the cochlear nuclei, optical density ratios in the 3.0 kHz region increased monotonically with increasing stimulus intensity, to a plateau at 45 dB SPL. In higher auditory nuclei, relative 2-DG uptake increased to a peak at 45 dB SPL and then declined at higher intensities.

Action Potentials↗
PubMed1989

Spiral ganglion cell density in young and old gerbils.

The Mongolian gerbil, like other mammalian species, has a decreased number of spiral ganglion cells as a function of age. This loss of cells was first seen in 24- to 30-month old animals in the basal end of the ganglion. In the oldest individuals the apical end of the ganglion was also affected. There were approximately 15-25% fewer cells in the affected areas in the 36- to 42-month old animals. In the oldest animals degeneration of the stria vascularis was seen in the apical turn and some degenerative changes in the organ of Corti were seen throughout the length of the cochlear duct. The aging pattern in the gerbil cochlea, is similar to that described for other species. Vacuoles, previously described in the gerbil cochlear nucleus, were also seen in the auditory nerve within the modiolus, but central to the Schwann-glial border in all animals. Vacuoles were not present within the spiral ganglion or the peripheral processes of the ganglion cells. Because the ganglion cell axons should be similar on either side of the Schwann-glial border, but the vacuoles were confined to the central nervous system, it is concluded that the degenerative process affects glial cells as opposed to neurons.

Aging↗
PubMed1989

Congenital cytomegalovirus labyrinthitis and sensorineural hearing loss in guinea pigs.

Cytomegalovirus (CMV) is the leading cause of human nonhereditary congenital deafness. The pathogenesis of congenital CMV infection in the auditory system is poorly understood and no suitable animal model is currently recognized. In this study primary maternal CMV infection in guinea pigs during the first or second trimester of pregnancy resulted in congenital infection in 64% of the offspring. Of the congenitally infected neonates, 28% had significant auditory deficits. Within the inner ear, CMV infection was localized in auditory nerve spiral ganglion cells. These findings indicate that congenital CMV infection of the guinea pig results in physiologic and anatomic neuropathology similar to that seen in human infection and provide the first experimental model for congenital CMV-induced sensorineural hearing loss.

Action Potentials↗
PubMed1989

Development of morphological and physiological changes in the cochlea induced by cytomegalovirus.

The effect of viral infection in the cochlea was investigated by inoculation of live cytomegalovirus or inactivated virus. Auditory thresholds were measured on the day of inoculation and on the terminal day. Two to 8 days following inoculation, the animals were killed and the cochleas were evaluated histologically. The compound nerve potential showed an increase in threshold prior to the cochlear microphonic, indicating the nerve was affected prior to the outer hair cells. All experimental cochleas contained inflammatory and cytomegalic inclusion cells and showed degenerative changes. The number of infected cells was small relative to the histopathology. Control cochleas had normal structure and function. The degeneration, therefore, might be mediated by inflammation as well as by the cytopathic effect of the virus. Viral infections, therefore, might be better managed with anti-inflammatory therapy in addition to antiviral agents.

Animals↗
PubMed1988

Contributions of the middle ear to the development of function in the cochlea.

The contribution of middle ear immaturities to the development of cochlear microphonic potential (CM) responses was studied throughout the ontogeny of auditory function in the Mongolian gerbil. CM produced by direct mechanical stimulation of the stapes was compared with CM generated by acoustic stimulation of the intact ear at various postnatal ages. The results indicated that during development acoustically generated CM reflects middle ear as well as inner ear maturational factors. With direct stapes driving, CM was first elicited at 10 days after birth (DAB), two days earlier than with acoustic stimulation. Controlling for the contributions of middle ear immaturity, maturation of the inner ear accounted for approximately a 75 dB improvement in CM thresholds between 10 and 18 DAB. Comparisons between the results obtained with the acoustic and stapes driving stimulation protocols suggested that the major maturational changes in the middle ear conduction apparatus occurred between 14 and 16 DAB. This period was associated with the final stages of resorption of middle ear mesenchyme and ossicular ossification. Before 16 DAB, acoustically evoked CM thresholds reflect approximately a 25 dB loss in sensitivity due to middle ear immaturity.

Acoustic Stimulation↗
PubMed1988

Development of tonotopic representation in the Mongolian gerbil: a 2-deoxyglucose study.

The spatial representation of frequency in the central auditory system of the neonatal gerbil was mapped with the 2-deoxyglucose (2-DG) autoradiographic technique. At 14 days after birth (DAB), pure tone stimulation produced recognizable patterns of 2-DG uptake. However, at this age, tone-induced areas of increased 2-DG uptake occurred at locations which in the adult respond to higher frequencies. The degree of shift in tonotopic representation was approximately two octaves. The normal adult tonotopic organization of auditory nuclei was achieved by 18 DAB, consistent with the rapid development of auditory function in the gerbil. The results suggest that the spatial distribution of frequency in the cochlea of neonatal animals is different from that in adults. Stimulus-evoked 2-DG uptake occurred first in brainstem auditory nuclei, and was observed in midbrain and forebrain auditory structures only at later ages. This is consistent with a sequential development of function in the central auditory pathway.

Acoustic Stimulation↗
PubMed1988

The spatial representation of frequency in the rat dorsal cochlear nucleus and inferior colliculus.

The spatial distribution of neural activity produced by tones was assessed in the rat dorsal cochlear nucleus (DCN) and inferior colliculus (IC), using the 2-deoxyglucose (2-DG) technique. Eight pure tones, spanning the range of reported single unit characteristic frequencies in the rat, were presented at 40 dB above behavioral threshold. The relationship between frequency of stimulation and location of neural activity within each nucleus was evaluated quantitatively. Based on the 2-DG uptake pattern across animals, a tonotopic axis in the transverse plane was defined for each nucleus. This axis transected the centers of regions of evoked 2-DG uptake for each frequency. There was an orderly relationship between stimulus frequency and the location of evoked neural activity along the axis. Each pure tone stimulus activated an approximately equal proportion of this axis, for all frequencies tested, in both the DCN and IC. This suggests the existence of equal 'spatial bandwidths, in rat central auditory structures, across its entire frequency range. Equal spatial bandwidths could facilitate signal analysis strategies which require interaction between neurons with closely-related CFs. In the horizontal plane, however, the proportion of stimulated tissue was not equal across frequency. High-frequency (greater than 8 kHz) tones produced increased neural activity along a much greater extent of the anterior-to-posterior axis of the IC than did low-frequency tones.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Ganciclovir prophylaxis for cochlear pathophysiology during experimental guinea pig cytomegalovirus labyrinthitis.

The effectiveness of the antiviral agent ganciclovir (9-[1,3-dihydroxy-2-propoxymethyl]guanine) against guinea pig cytomegalovirus was tested in vitro in guinea pig embryonic fibroblasts and in vivo in an experimental guinea pig cytomegalovirus labyrinthitis model. In vitro, ganciclovir completely prevented guinea pig cytomegalovirus infection of guinea pig embryonic fibroblasts at concentrations above 32.6 micrograms/ml. In vivo, antibody-negative animals had an average 17-dB elevation in their auditory nerve compound action potential thresholds (P less than 0.01, t test) and showed signs bilaterally of guinea pig cytomegalovirus labyrinthitis 8 days after intrathecal inoculation of virus. Ganciclovir administration starting 1 day before inoculation prevented the development of both cochlear histopathologic change and hearing loss. Guinea pig cytomegalovirus meningitis was observed in both the drug-treated and untreated groups. High-pressure liquid chromatography confirmed the presence of ganciclovir in the serum, perilymph, and cerebrospinal fluid of the drug recipients. Prophylactic ganciclovir thus can protect the cochlea from the histopathologic changes and hearing loss normally associated with experimental guinea pig cytomegalovirus labyrinthitis.

Acoustic Stimulation↗
PubMed1987

Pentobarbital and ketamine alter the pattern of 2-deoxyglucose uptake in the central auditory system of the gerbil.

Relative 2-deoxyglucose (2-DG) uptake was investigated during pentobarbital and/or ketamine anesthesia, when animals were either kept in silence or stimulated with wide band noise at 85 dB SPL. In the absence of anesthesia, noise stimulation produced a large increase in relative 2-DG uptake, when compared to silence, in all auditory nuclei up to and including the inferior colliculus. Much more modest noise-induced increases were seen in the medial geniculate nucleus and auditory cortex. These effects were markedly altered by anesthesia. Pentobarbital, and especially pentobarbital plus ketamine, enhanced stimulus-evoked increases in relative 2-DG uptake in lower auditory nuclei: the cochlear nuclei, superior olivary complex and ventral nucleus of the lateral lemniscus. At the same time, stimulus-evoked increases were decreased in the dorsal nucleus of the lateral lemniscus and inferior colliculus, and virtually eliminated in the medial geniculate and auditory cortex. The results of this study permit more meaningful comparison of 2-DG techniques with electrophysiological measures of central auditory activity, and illuminate the utility and limitations of each method. The data indicate that 2-DG observations from barbiturate-anesthetized preparations should be interpreted with some caution. They further suggest that the 2-DG technique is inappropriate for the study of stimulus-evoked activity in the medial geniculate and auditory cortex of barbiturate-anesthetized animals.

Acoustic Stimulation↗
PubMed1986

Development of mammalian endocochlear potential: normal ontogeny and effects of anoxia.

The development of the positive endocochlear potential (EP), the negative anoxic EP, and the organ of Corti potential were measured at various postnatal ages in the Mongolian gerbil, beginning at 8 days after birth (DAB). The organ of Corti potential (OCP) was present at 8 DAB but averaged 21% less than the adult value. OCP increased regularly with age, reaching adult values of -90 mV by 14 DAB. The positive EP was first observed at 10 DAB, at which age it averaged only 2-3 mV. This potential increased monotonically between 10 and 20 DAB, by which time it had reached the adult value of 75 mV. Anoxia did not result in a negative EP until 12 DAB, at which age this potential averaged -7 mV. The negative anoxic EP matured more rapidly than the positive EP, achieving the adult value of 40 mV by 18 DAB. During development the positive EP appeared to closely parallel the maturation of glucose metabolism in the stria vascularis. The negative anoxic EP was more closely related temporally to the development of cochlear microphonic potential (CM) thresholds. It is hypothesized that changes which occur between 10 and 16 DAB in the apical membranes of the cochlear hair cells contribute to the maturation of both CM and the negative anoxic EP.

Animals↗
PubMed1985

Ventral cochlear nucleus neural discharge characteristics in the absence of outer hair cells.

The role of the cochlear outer hair cell (OHC) in auditory processing remains poorly understood. The OHCs possess an independent afferent innervation which constitutes 5-10% of cochlear afferent neurons and which appears to project to the cochlear nucleus (CN). Whether the OHCs contribute to the processing of auditory signals in the CN has not been determined. To address this question, kanamycin ototoxicity was used to produce selective OHC loss while leaving the inner hair cell (IHC) population largely intact, in the basal portion of the cochlea of chinchillas. Single unit responses were then recorded in the ventral cochlear nucleus (VCN), and compared to responses in untreated subjects. Many of the changes observed in VCN neural responses reflected changes which have previously been reported in the VIIIth nerve. However, frequency tuning curve tip segments which were normal in both bandwidth and length were observed in approximately 22% of the units associated with regions of complete OHC loss and preservation of IHCs. This has not been reported in previous OHC lesion studies. Also, first spike latency was found to be significantly lengthened for units associated with the OHC free regions. Those features of VCN neural responses which first arise within the CN, such as non-primary-like post-stimulus-time histogram response patterns, were unaffected by OHC loss. These results suggest that afferent fibers associated with OHCs do not play a major role in signal processing in the VCN.

Animals↗