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Biomedical subjects

K E Rarey

Publications and source records attributed to K E Rarey.

At least 37 records · Page 2Linked to original sources

De novo synthesis of glucocorticoid hormone regulated inner ear proteins in rats.

Changes of rat inner ear de novo protein synthesis in response to dexamethasone (DEX), a synthetic glucocorticoid, have been analyzed by high resolution two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (2D-SDS-PAGE) and fluorography. Two proteins (M(r) 41,000 and 35,000) were amplified and one protein (M(r) 47,000) was suppressed by DEX in a cochlear culture medium. In the culture medium conditioned by vestibular tissue, three proteins (M(r) 67,000, 57,000 and 50,000) were amplified after DEX administration. In cochlear and vestibular tissues, glucocorticoid-responsive protein synthesis was down-regulated by DEX, including two proteins (M(r) 39,000 and 35,000) in the cochlea and five proteins (M(r) 80,000, 64,000, 59,000, 56,000 and 40,000) in the vestibule. The regulation of these inner ear proteins by DEX suggests that glucocorticoid may play an important role in normal inner ear microhomeostasis, as well as in the treatment of some inner ear disorders.

Animals↗

Glucocorticoid receptor expression in the postnatal rat cochlea.

The glucocorticoid receptor (GR) expression in the neonatal rat cochlea was investigated by utilization of a polyclonal antibody against GR, the immunoreactivity of which exhibited a distinct, age-dependent developmental pattern in tissues of the spiral ligament (SL). Immunostaining of GR appeared initially at the 7th postnatal day (PND), increased rapidly between the 14th and 21st PND, and reached adult-like expression levels by the 21st PND. Less pronounced, developmentally regulated expression patterns of GR were observed in cells of the spiral limbus (SLi), spiral ganglion (SG), organ of Corti (OC), and cochlear nerve (CN). For example, high expression levels of GR were observed in the SLi, SG and OC at 3 PND; subsequently, GR immunoreactivity levels decreased from 7 to 14 PND, and then GR immunoreactivity intensified in these regions by 21 PND. No remarkable changes in GR expression were observed in stria vascularis (SV). These data indicate that GR expression in the inner ear is tissue and age-specific, and that GR expression parallels both Na,K-ATPase expression and endocochlear potential development.

Aging↗

Combined effects of adrenalectomy and noise exposure on compound action potentials, endocochlear potentials and endolymphatic potassium concentrations.

The effects of removal of endogenous corticosteroids via bilateral adrenalectomy in combination with noise exposure (30 min at 100 dB) were determined by recording compound action potential (CAP) and endocochlear potentials (EP), and by measuring potassium concentrations (K+e) within the endolymph. Thirty-eight Long-Evans rats were divided into groups according to experimental treatments: adrenalectomy (ADX) or non-ADX and noise exposure or non-noise exposure. CAP thresholds, EP and K+e values were subjected to repeated-measures analysis of variance with group and time as factors classifying the measurements. Noise exposure resulted in significant elevations of CAP thresholds in both the ADX and non-ADX animals, but had no effect on either EP or endolymphatic K+e. Recovery was noted during all post-exposure measurement periods and was significantly faster for ADX animals. EP and K+e did not change during or after noise exposure. ADX animals showed a non-significant reduction of EP and a statistically significant increase of K+e during all measurement periods as compared to non-ADX animals.

Acoustic Stimulation↗

Effect of stress on cochlear glucocorticoid protein. II. Restraint.

The effect of restraint stress via immobilization on rat cochlear glucocorticoid receptor (GR) levels was determined using an enzyme-linked immunosorbent assay (ELISA). Results demonstrated that GR levels in cochlear tissues exhibited tissue-specific and time-dependent responses to immobilization (6 hours daily). Similar responses of the GR were observed in rats restrained during two different times of the day. A significant quadratic trend (P = 0.019, R2 = 0.58) was observed in levels of GR in spiral ligament tissues of rats restrained from 10:00 to 16:00 h; levels of GR were elevated by day 2, and by day 21 GR levels had returned to near normal levels. GR levels in the spiral ligament tissues also were found to increase significantly after 2 days in response to repeated restraint stress administered from 06:00 to 12:00 h (P = 0.017, R2 = 0.34). Interestingly, a subtle, but statistically significant, decreasing trend in the organ of Corti's GR levels was detected when the daily restraint stress was applied from 06:00 to 12:00 h for up to 7 days. No significant trends (P > 0.05) were observed in GR levels of stria vascularis tissues regardless of the time of day of the restraint protocol. Stress has been implicated as an etiological factor in Ménière's disease and other ear pathologies. The data presented here indicate that the effect of stress is specific to tissue region and that, as in tissues of other systems, the GR of cochlear tissues are responsive to stress.

Analysis of Variance↗

Clinical viral infections and temporal bone histologic studies of patients with AIDS.

The brain, eye, and inner ear are each protected from blood-borne infectious agents by a barrier that has some anatomic and functional differences. In patients with AIDS, opportunistic infections of the central nervous system and eye are frequent. Little is known about the incidence of middle and inner ear infections in patients with AIDS, but deafness and severe vertigo are uncommon. We studied 14 homosexual men with AIDS, aged 28 to 55 years, for 1 to 2 years until death. No patient had deafness, but one had vertigo. Adenovirus type 6 and cytomegalovirus were isolated from the middle ear cavity in four patients. Temporal bone histology demonstrated acute otitis media in four, chronic otitis media in two, and serous otitis media in three. Adenovirus type 6 and cytomegalovirus, either alone or with herpes simplex virus type 1, were isolated from inner fluids of three patients. Histologic inner ear findings were abnormal in only one patient. Viruses were isolated or histologically identified in the brains of four patients and in the eyes of five patients. In our patients viral infections were nearly as common in the inner ears as in the brain and eye, suggesting that protection from the blood-labyrinth barrier was similar to that from the other barriers. Because the inner ear viral infections were asymptomatic and there was an absence of pathologic damage and inflammation, we suggest that some viral inner ear infections in patients with AIDS are nonpathogenic and elicit no inflammation or that the viral infections occur terminally and elicit no inflammation because of immunosuppression from the AIDS.

AIDS-Related Opportunistic Infections↗

Expression of Na, K-ATPase alpha and beta isoforms in the neonatal rat cochlea.

The postnatal expression of five Na, K-ATPase alpha (alpha 1, alpha 2, alpha 3) and beta (beta 1, beta 2) subunit isoforms in the rat cochlea was investigated by immunocytochemistry. High levels of expression of the alpha 1 and beta 2 isoforms were observed in stria vascularis (SV) at all developmental stages. alpha 1 and beta 1 isoforms showed a distinct time-dependent developmental expression pattern in tissues of the spiral ligament (SL) and spiral limbus (SLi). Limited, temporary expression of alpha 2 and alpha 3 subunit isoforms were found in SV and SL. Expression of each isoform was also seen in organ of Corti (OC), spiral ganglion (SG), cochlear nerve (CN) and Kölliker's Organ (KO). These observations suggest that individual isoforms may exert specific actions postnatally during final cochlear maturation.

Animals↗

Effects of low-sodium, high-potassium dietary intake on cochlear lateral wall Na+,K(+)-ATPase.

The effect of a low Na+, high K+ diet on Na+,K(+)-ATPase levels in cochlear lateral wall tissues was investigated in laboratory rats by using an enzyme-linked immunosorbent assay. The low Na+, high K+ diet induced high aldosterone plasma levels in the animals as well as changes in plasma cation levels. Animals that received a low Na+, high K+ diet demonstrated a statistically significant (97%) increase in Na+,K(+)-ATPase levels in the stria vascularis when compared to animals that received a control diet. This increase in strial Na+,K(+)-ATPase levels was blocked only 70% by administration of the aldosterone antagonist, spironolactone. Findings therefore indicate that strial Na+,K(+)-ATPase may be modulated by both aldosterone and Na+,K+ plasma levels. Na+,K(+)-ATPase levels in the spiral ligament were not affected by the experimental treatment. These findings suggest that spiral ligament Na+,K(+)-ATPase levels may be regulated by factors other than aldosterone and Na+,K+ plasma levels. This study provides further insight into the mechanisms of the beneficial effects of salt restriction and potassium loading in patients with Méniére's disease.

Aldosterone↗

Expression of Na+,K(+)-ATPase alpha 1 subunit mRNA in the developing rat cochlea.

The distribution of Na+,K(+)-ATPase alpha 1 subunit mRNA was identified using in situ hybridization in the developing rat cochlea. The expression of alpha 1 subunit mRNA in stria vascularis (SV) was observed in all time points studied, 1 to 30 postnatal day (pnd) rats. The adult expression level was attained between 11 to 14 pnd. Surprisingly, alpha 1 subunit mRNA in spiral ligament (SL) and spiral limbus (SLi) was expressed in a more distinct time-dependent manner. At 7 pnd, the alpha 1 subunit mRNA expression was observed initially in the tissues of the SL. At 11 pnd, alpha 1 subunit expression appeared in SLi. Between 11 and 14 pnd, an adult-like pattern of Na+,K(+)-ATPase alpha 1 subunit mRNA expression was attained in the SL and SLi. These data suggest that the expression of Na+,K(+)-ATPase alpha 1 subunit mRNA in these areas are closely related to the development of the rat EP, as its expression in the stria vascularis.

Animals↗

Na,K-ATPase alpha and beta subunit isoform distribution in the rat cochlear and vestibular tissues.

The distribution of five Na,K-ATPase subunit isoforms (alpha 1, alpha 2, alpha 3, beta 1 and beta 2) in rat cochlear and vestibular tissues was determined by immunocytochemical techniques using subunit isoform specific polyclonal antibodies. The expression of Na,K-ATPase alpha and beta subunit isoforms varied among different cell regions of the inner ear. The alpha 1 subunit isoform was more extensively distributed in all inner ear tissues than the alpha 2 or alpha 3 subunit isoforms. The beta 1 subunit isoform was distributed primarily in spiral ligament and inner hair cells of the cochlea, and in crista ampullaris and macula of the saccule. The beta 2 subunit isoform was most abundant in the stria vascularis, dark cells of the ampullae and utricle. The alpha 1 beta 1 subunit combination of Na,K-ATPase was most commonly found in the spiral ligament, while the alpha 1 beta 2 combination was most abundant in the stria vascularis. Similarly, alpha 1 beta 2 was confined more to the dark cells of the ampullae and utricle. The alpha 3 beta 1 subunit combination of Na,K-ATPase was identified in the inner hair cells of the cochlea and the sensory regions of the vestibular end organs. These observations may reflect functional diversity of Na,K-ATPase in the individual inner ear regions and may provide insight into the differences between fluid and ion transport in the inner ear and that of other transporting tissues. Overall, the distribution pattern further indicates that the different isoform combinations have specific roles.

Animals↗

11 beta-Hydroxysteroid dehydrogenase in the rat inner ear.

11 beta-Hydroxysteroid dehydrogenase (11 beta-HSD) was demonstrated specifically in the spiral ligament of the cochlear membranous labyrinth by enzyme assay, Western blotting, and immunocytochemistry. Other cochlear regions and the vestibular membranous labyrinth were devoid of 11 beta-HSD. Spiral ligament 11 beta-HSD exerted predominantly an oxidative activity and was NADP specific, which is similar to 11 beta-HSD in most other tissues. 11 beta-HSD was colocalized with mineralocorticoid and glucocorticoid steroid receptors in the spiral ligament. 11 beta-HSD may control steroid binding to these inner ear steroid receptors and, in addition, may regulate steroid receptor binding in the adjacent stria vascularis in paracrine fashion.

11-beta-Hydroxysteroid Dehydrogenases↗

Density of marginal cell basolateral membranes in basal, middle, and apical turns of the rat cochlea.

The Na-K-ATPase activity within stria vascularis tissues has been observed to decrease from basal to apical cochlear turns. This reduction in Na-K-ATPase activity in more apical regions of the cochlea may be accounted for, at least in part, by a lower density of Na-K-ATPase-containing basolateral membranes of the marginal cells. This study was performed to compare the surface density of marginal cell basolateral membranes in standardized basal, middle, and apical turns of the rat cochlea. Montages of transmission electron microscopic micrographs of standardized regions of the rat stria vascularis were morphometrically analyzed. Surface area of marginal cell basolateral membrane was determined per unit stria vascularis volume and per unit marginal cell volume. Results demonstrated that the surface density of marginal cell basolateral membrane per stria vascularis volume of the standardized basal region increased 20 percent compared to that of the standardized apical region (p < or = .1). Surface density of marginal cell basolateral membranes per marginal cell volume was similar in all three standardized turns. The observed longitudinal differences in surface density of marginal cell basolateral membranes per stria vascularis volume may account, in part, for the biochemical cochlear Na-K-ATPase activity gradient from base to apex.

Animals↗

Dynamics of Na,K-ATPase sites in lateral cochlear wall tissues of the rat.

The objective of this study was to determine whether varying levels of either the synthetic glucocorticoid, dexamethasone, or the mineralocorticoid, aldosterone, modulate the quantity of Na,K-ATPase sites in stria vascularis and spiral ligament tissues. Surgically adrenalectomized male rats were administered different dosages of dexamethasone or aldosterone for 7 days and subsequently were sacrificed. Na,K-ATPase alpha-subunits of stria vascularis and spiral ligament homogenates were determined quantitatively by enzyme-linked immunosorbent assay, using a monoclonal antibody shown to react with lateral wall Na,K-ATPase alpha-subunit. Elevated serum levels of dexamethasone were correlated with significantly increased Na,K-ATPase alpha-subunit levels in both the stria vascularis and spiral ligament (P < 0.05). Elevated serum levels of aldosterone were correlated with increased Na,K-ATPase alpha-subunits in the stria vascularis, but not in the spiral ligament (P < 0.1). These data further indicate a positive correlation between increased serum levels of adrenal steroids and induction of Na,K-ATPase synthesis by such steroids, particularly by dexamethasone.

Adrenalectomy↗

Tissue specific levels of glucocorticoid receptor within the rat inner ear.

Individual, rat inner ear tissues were isolated and processed for determination of levels of glucocorticoid (GR) receptor by an Enzyme Linked Immuno-Sorbant Assay (ELISA). Differing levels of GR receptor between seven sampled inner ear regions were measured. Levels of GR receptors in the spiral ligament tissues were found to be significantly higher compared to all other tissue samples. GR levels in the tissues of stria vascularis and organ of Corti were different from one another but both were statistically higher than those detected in the vestibular tissue samples (dark cell regions, cristae ampullares and maculae utriculi), which had the lowest GR receptor levels measured. Intermediate levels of GR receptor were found in the endolymphatic sac region. It is suggested that the varying levels of inner ear GR receptors may be indicative of differing biological responses among the given tissues, as well as differences in the magnitudes of such responses to circulating glucocorticoids.

Analysis of Variance↗

Localization of glucocorticoid receptors and glucocorticoid receptor mRNAs in the rat cochlea.

The distribution of glucocorticoid (GR) receptor messenger RNAs (mRNAs) and GR receptors was studied by in situ hybridization histochemistry and immunocytochemistry, respectively. In situ hybridization histochemistry was performed with a biotin-labeled riboprobe complementary to rat GR receptor mRNA. GR receptor mRNAs were demonstrated in spiral ligament cells, spiral limbus cells, and spiral ganglion cells. GR receptor mRNAs were demonstrated neither in cells of the stria vascularis nor in cells of the organ of Corti region. With the use of a monoclonal and a polyclonal antibody, GR receptors were observed in the spiral ligament cells, stria vascularis cells, spiral limbus cells, and spiral ganglion cells by immunocytochemistry. Binding of anti-GR-receptor antibodies to a lesser extent was observed in the organ of Corti region; however, cellular distribution of the GR receptors could not be resolved with the applied techniques. These results suggest that the GR receptor is expressed differently in the heterogeneous cochlear tissues.

Animals↗

Temporal bone histopathology 14 years after cytomegalic inclusion disease: a case study.

Temporal bones were examined from a 14-year-old male who died of sequelae of congenital cytomegalic inclusion disease (CID). Cytomegalovirus (CMV) was not isolated from inner ear fluid or multiple systemic tissues at the time of death. Examination of temporal bones revealed chronic pathology of both cochlear and vestibular sensory and nonsensory tissues. Endolymphatic hydrops was observed in the basal turn of the cochlear duct, while Reissner's membrane was collapsed in the more apical turns. Strial atrophy and a loss of cochlear hair cells were observed along the entire length of the basilar membrane. Vestibular neuroepithelial regions were degenerated and fibrosis was seen within the vestibular perilymphatic tissue spaces, suggesting prior labyrinthitis within the perilymph compartment in addition to the more typical pattern of endolabyrinthitis associated with human CMV infection. Distention of the saccular membrane was evident. In both cochlear and vestibular tissues, there were isolated regions of calcifications that appeared characteristic to that reported in other organ systems of individuals with CID. Collectively, these chronic, pathological findings in this case of CID demonstrate more extensive injury than has been identified in the previously reported acute temporal bone pathology of CID.

Cochlea↗

Immunochemical detection of glucocorticoid receptors within rat cochlear and vestibular tissues.

A monoclonal antibody, BuGR2, to liver glucocorticoid receptor sites was tested for its reactivity and specificity in inner ear tissue supernatants by an Enzyme Linked Immuno Sorbant Assay (ELISA) and a Western blotting technique. Results demonstrated that this antibody specifically recognized a protein of 93 kDa in inner ear supernatant fractions, which conformed to the reported molecular weights of the glucocorticoid receptor in other tissues. Antigenic sites were determined to be higher within cochlear supernatant fractions compared to vestibular supernatant fractions by ELISA. This anti-glucocorticoid receptor antibody combined with the quantitative ELISA provides a sensitive means to further investigate the inner ear glucocorticoid receptor system.

Animals↗

Planum semilunatum of the rat: new light and electron microscopy observations.

Planum semilunatum (PSL) cells of the rat ampullae were studied by light, transmission, and scanning electron microscopy. The observed ultrastructure of rat PSL cells was similar to that described in other species, but is in disagreement with earlier reports of rat PSL cells, most probably because of previous divergent definitions of the PSL. Regions of PSL cells were easily distinguished from other nonsensory epithelia at the lateral ends of the crista ampullaris. The PSL region consisted of irregular-shaped columnar to cuboidal pentagonal or hexagonal cells that interdigitated with one another by lateral membrane infoldings. In the PSL region the subepithelial reticular layer appeared thickened and formed wartlike impressions in the basal surface of the PSL cells. These morphological characteristics of the subepithelial reticular layer were unique to the PSL region in the ampulla and may reflect special adaptations of the PSL region to mechanical stress. Furthermore, the thick subepithelial reticular layer may have implications for transport across the PSL region.

Animals↗

Plasma membrane modulation of ampullar dark cells by corticosteroids.

Individual effects of corticosteroids on the ultrastructure of rat ampullar dark cells were quantitatively determined. Adrenalectomized rats subsequently received either aldosterone or dexamethasone or only hormone solvent via miniosmotic pumps for a period of 14 days. Ampullar tissues of the animals were processed for transmission electron microscopy, and standardized regions of ampullar dark cells were photographed and morphometrically analyzed. Surface density and boundary length of basolateral membrane increased significantly after hormone substitution with aldosterone, but not after replacement with dexamethasone, as compared with animals that received only solvent. No significant differences were observed for other morphological parameters between the groups. This response of ampullar dark cells to aldosterone appears similar to that of principal cells of the collecting duct in comparable experimental conditions. These morphological observations suggest that the mineralocorticoid, aldosterone, modulates ampullar dark-cell membranes.

Adrenal Glands↗