Pigmented cells of the stria vascularis and spiral ligament of the chinchilla

This study was designed to investigate the cellular distribution and ultrastructure of melanin pigment in the stria vascularis and spiral ligament of the chinchilla cochlea. Typical dendritic melanocytes containing homogeneously distributed eumelanin granules were observed in the spiral ligament. In the stria vascularis melanin was found to occur in three types of cells (heavily pigmented cells that appear to be melanocytes, intermediate cells and basal cells). The melanocyte-like cells contain pigment globules composed of melanin granules, granular matrix and occasional lipid droplets in a configuration similar to that of neuromelanin. These cells are morphologically distinct from intermediate cells which contain sparsely distributed, small, dense granules apparently composed of melanin. The intermediate cells show a positive DOPA reaction and portions of the intermediate cell GERL system display intense acid phosphatase reactivity. The basal cells of the chinchilla stria were also found to contain occasional clusters of melanin granules. It is hypothesized that the basal cells may acquire melanin by donation from the other pigmented cells of the lateral cochlear wall.     

Ultrastructural localization of Ca2+-binding sites in the spiral limbus,the stria vascularis and Reissner's membrane of the guinea pig

Summary Cations were precipitated with potassium antimonate in the cochlea of the guinea pig and electron microscopy was used to analyze the distribution of the formed reaction products. Differences in precipitate density between neighboring cells in Reissner's membrane, in the stria vascularis and in the limbus are described. Electron spectroscopic imaging was performed to obtain information about the spatial distribution of the precipitates and their elemental composition.     

Ultrastructural localization of Ca2+-binding sites in the spiral limbus, the stria vascularis and Reissner's membrane of the guinea pig

Cations were precipitated with potassium antimonate in the cochlea of the guinea pig and electron microscopy was used to analyze the distribution of the formed reaction products. Differences in precipitate density between neighboring cells in Reissner's membrane, in the stria vascularis and in the limbus are described. Electron spectroscopic imaging was performed to obtain information about the spatial distribution of the precipitates and their elemental composition.     

Susceptibility of organ of Corti with or without melanin to acoustic overstimulation]

Albino and pigmented guinea pigs were compared in terms of susceptibility to acoustic trauma. The animals were exposed to a 4 kHz pure tone of 120 dB for 60 min. N1 thresholds of CAP were measured before and after the acoustic exposure. Changes in the outer hair cell and stria vascularis were studied using SEM and TEM. After acoustic trauma, N1 thresholds were more elevated in the albino than in the pigmented guinea pigs. Also, pathological changes in the outer hair cell and stria vascularis were more severe in the albino animals. A noteworthy finding in the stria vascularis was that the melanin in intermediate cells had moved into marginal cells. This melanin migration may be possibly involved in mechanisms underlying prevention of acoustic trauma.     

Alteration of loosely bound calcium in the guinea pig organ of Corti after treatment with diltiazem as calcium channel blocker

After oral administration of the organic calcium channel blocker diltiazem to guinea pigs for 7 days, calcium ions were precipitated with potassium antimonate in the cochleae. The spatial distribution of the precipitates was studied by energy-filtering transmission electron microscopy and the amount of the ultrastructural reaction products formed was determined semiquantitatively by an image processing system. Compared with untreated control ears, the number of the formed precipitates was reduced drastically in the inner hair cells after diltiazem treatment. In addition, electron microscopic analysis revealed that the number of calcium precipitates attached at the basolateral membrane of the outer hair cells was clearly reduced when compared with untreated control specimens. A large number of histochemical reaction products could be identified in the basilar membrane and were also observed in the untreated control specimens. The spatial distribution of the calcium precipitates in the lamina reticularis was not affected by diltiazem treatment and calcium precipitates could be identified within different cell membranes. The technique used was considered to be helpful for identifying calcium channels ultrastructurally in intact undissected tissues and to support light microscopic analyses and patch-clamp electrophysiological measurements.     

庆大霉素对豚鼠血管纹黑色素的影响及其机制

OBJECTIVE: To investigate the effect and its mechanism of gentamicin(GM) on melanin in stria vascularis of guinea pig. METHODS: The differences of auditory thresholds between pigmented and albino guinea pigs, given GM of 150 mg/kg for 7 days, were studied. Moreover, the content of melanosomes, activity of tyrosinase and expression of proliferating cell nuclear antigen(PCNA) in intermediate cells of stria vascularis in gentamicin-treated pigmented guinea pigs were compared with those in control animals by electron microscope and immunohistochemistry, respectively. RESULTS: After gentamicin exposure, the auditory thresholds of all animals increased significantly (P < 0.001), whereas threshold shifts averaged across all frequencies of pigmented animals were much less than those of the albinos(P < 0.001). The number of melanosomes of each examined area (300 microns 2) in intermediate cells was obviously increased from 19.83 +/- 2.74 to 58.33 +/- 16.22. The ratio of tyrosinase reaction products area to the total measured area was significantly increased from 1.65% +/- 0.40% to 3.45% +/- 0.41% after gentamicin exposure. However, the numbers of positive intermediate cells expressing PCNA were 14.08 +/- 2.76 and 13.58 +/- 2.09 before and after gentamicin treatment, respectively. But there was no statistically significant difference between them (P > 0.05). CONCLUSION: The increase of content of melanin in stria vascularis after GM exposure does not result from the change of proliferating activity of melanocytes, but from the enhanced tyrosinase activity. Melanins in stria vascularis may possess the ability to protect the inner ear from ototoxicity of gentamicin.     

The development of melanin in the stria vascularis of the gerbil.

One factor that influences noise susceptibility is pigmentation. The aim of this study was to investigate the development of melanocytes, other melanin-containing cells and the amount of melanin in stria vascularis from birth to adult age in the gerbil which has a uniform pigmentation of the fur and eyes, is born without hearing but establishes hearing function at 14-18 days after birth. Changes in the melanin morphology, concentration and distribution have been correlated to the development of the inner ear and to the time period during which hearing function is established, which indicates that the melanocytes in stria vascularis are of importance for the hearing function.     

Characteristics of stria vascularis melanocytes of viable dominant spotting (Wv/Wv) mouse mutants.

The Wv mutation lies in the kinase domain of the proto-oncogene c-kit which is expressed in a variety of cells including neural crest derived melanoblasts. The mutation results in the abnormal migration, proliferation, survival and/or differentiation of melanoblasts. Viable Dominant Spotting (Wv/Wv) mouse mutants have a white coat due to the absence of melanocytes. The majority of these animals have no melanocytes within the stria vascularis and no endocochlear potential (EP). A proportion of homozygous mutants partially escape the effects of the mutation: 47.2% of pinnae and 21% of vestibular regions were pigmented and 10.8% of ears had an EP. All ears with an EP that were available for histology had some pigmentation of the stria. There was no obvious correlation between external and internal spotting in Wv/Wv mice, and asymmetrical pigmentation of the ears was common. Both light and dark intermediate cells (which are derived from melanocytes) were present in the middle and/or basal turns of these cochlear ducts and they appeared to function normally in enabling the stria to produce an EP (although the EP was usually lower than normal). This suggests that the c-kit gene product is needed only during development of the stria, and not for mature melanocyte function because the melanocytes present in the mutant strias were carrying the mutant version of the c-kit gene. Melanocytes were similar in appearance in controls and mutants, except that fewer melanin granules were observed in the strias of Wv/Wv mice. The observations that strial melanocytes with very few melanin granules in Wv/Wv mutants are able to support EP production, together with previous observations that albino animals with strial melanocytes but no melanin have a normal EP, suggest that melanocytes but not melanin are essential for normal strial function.     

Comparison of the quantity of cochlear melanin in young and old C57BL/6 mice

OBJECTIVE: To elucidate the functional relationship between cochlear melanin and aging. DESIGN: Melanin has been described in the cochlear labyrinth and has been suggested to protect the cochlea from various types of trauma. The quantity of melanin has been shown to change with aging in several organs; however, to our knowledge, aging changes in the cochlea have not been documented. Therefore, we chemically quantified cochlear eumelanin and pheomelanin contents and compared these in young and old C57BL/6 mice using high-performance liquid chromatography. Because melanin deposits in the cochlea present most extensively in the stria vascularis, we morphologically examined the stria using transmission electron microscopy. SUBJECTS: Cochleae from an inbred strain of C57BL/6 male and female mice; 6 at the age of 10 weeks and 5 at the age of 100 weeks were studied. RESULTS: The quantities of cochlear eumelanin and pheomelanin were 421 and 480 ng per cochlea in young mice, and 2060 and 765 ng per cochlea in old mice, respectively. Under transmission electron microscopy, the number of pigmented granules seemed to be greater in older mice compared with younger mice, especially in marginal cells. CONCLUSION: To our knowledge, our findings are the first quantitative evidence to show an age-related overexpression of cochlear melanin and an alteration in the proportion of eumelanin and pheomelanin with aging, suggesting a possible otoprotective function of eumelanin against age-related cochlear deterioration.     

内耳色素对爆震性听损伤保护作用的实验观察

目的　探讨内耳色素与爆震性听损伤的关系。方法　利用耳蜗铺片、石蜡切片、透射电镜及扫描电镜观察爆震前后白化豚鼠和杂色豚鼠耳蜗形态结构的变化 ,并对爆震前后白化豚鼠和杂色豚鼠听性脑干反应(ABR)反应阈进行测定。结果　白化豚鼠的耳蜗形态损伤和听功能损伤均较杂色豚鼠严重。结论　爆震后白化豚鼠耳蜗形态学损伤及ABR反应阈的改变均较杂色豚鼠明显。提示内耳血管纹色素颗粒与爆震性内耳损伤有关 ,机理可能为色素颗粒参与调节爆震后内淋巴液中钙离子浓度的平衡及参与清除爆震后耳蜗产生的氧自由基     

鼠尾胶原在原代培养耳蜗血管纹边缘细胞中的应用

目的：建立利用自制的鼠尾胶原培养大鼠耳蜗边缘细胞的方法。方法：制作鼠尾胶原,并观察利用自制的鼠尾胶原所培养出大鼠耳蜗边缘细胞的效果。结果：自制的鼠尾胶原能正常地培养出大鼠耳蜗边缘细胞,细胞角蛋白18表达阳性,免疫组织化学结果和扫描电镜证实所培养的细胞具有典型的分泌上皮细胞特征。结论：成功建立了利用自制鼠尾胶原培养大鼠耳蜗边缘细胞的方法,并且制作简便,成本低廉。     

Noise-induced hearing loss: the effect of melanin in the stria vascularis.

Conflicting investigations regarding the potential protective effect of melanin against noise-induced sensorineural hearing loss have suggested that eumelanin and pheomelanin may have differing effects within the stria vascularis. Three strains of C57BL/6J mice, (+/+, a/a) wild-types (dark coats/black eyes), (c2j/c2j, a/a), albinos (white coats/pink eyes), and (+/+, Ay/Ay) yellow mice (yellow coats/black eyes), were subjected to five consecutive days of broad band noise exposure at 112 dB(A) SPL for 3 h/day. Cochlear function was evaluated with auditory brainstem response audiometry to pure tones immediately pre-exposure, 5-6 h postexposure, and 14 days post-exposure. No significant difference in the degree of sensorineural hearing loss induced in the three strains of mice was identified. The eumelanin and pheomelanin content of each stria vascularis and amount of protein per stria for both mouse and guinea pig (2/NCR) were determined via high performance liquid chromatography. No pheomelanin was found in the stria of yellow mice, suggesting that coat color is not an accurate predictor of strial melanin content. The melanin content per mg of strial protein was higher in mice than in guinea pigs. A species-specific difference in melanin content does not explain the absence of a protective effect in mice.     

Cochlear degeneration in aged rats of four strains.

Animals with various degrees of inbreeding, some of which are albino, are frequently used in biological research. Albinos do not produce melanin and it is therefore absent from the cochlea. While the function of melanin is unknown, it has been hypothesized that it is involved in cochlear homeostasis. It is possible then, that age-related degeneration may be affected by the presence or absence of melanin. We therefore evaluated young (2-6 months old) and aged (24-36 months old) cochleas in 4 different rat strains: albino Fischer 344 and Lewis rats and pigmented Lewis-Brown Norway F1 rats and Brown Norway rats. Cochlear morphology was the same across all strains of young adult animals with the exception that the pigmented animals had small, darkly stained granules in the stria vascularis. The aged pigmented animals all had large granules as well as small ones. Degeneration of spiral ganglion cells in the apical region of the ganglion had occurred in the old animals of all strains. Strial degeneration at the apex was also present in aged animals. There was no correlation between the presence or absence of melanin and the magnitude of cochlear degenerative changes in the aged animals. The presence or absence of melanin therefore, appears to have no effect on cochlear degeneration in the aged rat cochlea.     

Spiral ligament and stria vascularis changes in cochlear otosclerosis: effect on hearing level.

OBJECTIVE: To investigate the effect of changes within the spiral ligament and stria vascularis on hearing in cochlear otosclerosis, we examined spiral ligament hyalinization, stria vascularis atrophy, and sensory hearing loss in cochlear otosclerosis and described changes in ion transport molecule expression. STUDY DESIGN: Retrospective. SETTING: Tertiary referral center. PATIENTS: Thirty-two cochleae from 24 temporal bone donors with histologic evidence of cochlear otosclerosis, including spiral ligament hyalinization. INTERVENTION: Audiography. MAIN OUTCOME MEASURES: Measurements of spiral ligament width, stria vascularis, and bone-conduction thresholds were compared by the amount of hyalinization. Expression of the ion transport molecules Na,K-ATPase, connexin 26, and carbonic anhydrase II were assessed by immunohistochemical techniques. RESULTS: Hyalinization most often involved the posterior basal turn (88%) and the posterior middle turn (27%). Spiral ligament hyalinization correlated significantly with stria vascularis atrophy in the posterior middle turn of the cochlea (rho = -0.63, p < 0.01). There was a trend toward a significant association in the posterior basal turn (rho = -0.31, p < 0.08). Bone-conduction thresholds at 2,000 and 4,000 Hz were significantly associated with the amount of stria vascularis atrophy (rho = -0.44, -0.40, p < 0.05). In addition, we observed decreased immunostaining for both carbonic anhydrase II with Type I fibrocytes and Na,K-ATPase with stria vascularis and Type II and Type IV fibrocytes of the spiral ligament in cochlear otosclerosis sections compared with normal cochlea. Na,K-ATPase staining within the stria vascularis was further decreased in the presence of spiral ligament hyalinization. No significant differences were seen with connexin 26 immunostaining. However, immunostaining results were somewhat inconsistent. CONCLUSION: These data suggest that spiral ligament structure and function are essential for stria vascularis survival. In addition, dampened expression of ion transport molecules within the spiral ligament and stria vascularis may disrupt potassium ion recycling, resulting in loss of endocochlear potential and sensory hearing loss.     

Effects of gadolinium injected into the middle ear on the stria vascularis

CONCLUSION: The concentration of gadolinium (Gd) used clinically showed no remarkable effects on the stria vascularis; however, a higher concentration had adverse effects. The concentration of Gd must be borne in mind when injecting Gd into the tympanic cavity. OBJECTIVE: Endolymphatic hydrops has been visualized using high resolution MRI with the intratympanic administration of Gd in patients with Meniere's disease. We attempted to investigate the effects of Gd on the stria vascularis. MATERIALS AND METHODS: Gd hydrate diluted eightfold with saline or non-diluted Gd or saline was injected into the tympanic cavity of guinea pigs. To investigate the effects of Gd on the stria vascularis, we measured endocochlear DC potential (EP) and observed the stria vascularis using transmission electron microscopy. RESULTS: Intratympanic injections of Gd hydrate diluted eightfold with saline (1/8 Gd) and saline did not cause apparent changes in the EP. Moreover, the amplitude of the EP decreased significantly 60 min after non-diluted Gd was injected. Transmission electron micrographs of the stria vascularis revealed no significant morphological difference between the ears injected with 1/8 Gd and those injected with saline. There was significant morphological change in the ear injected with non-diluted Gd. The intercellular spaces were markedly enlarged.     

Results of an ultrastructural study comparing stria vascularis with organ of Corti in guinea pigs treated with kanamycin

Ultrastructural study of ototoxicity is well documented with two points of interest: organ of Corti for aminoglycosides and stria vascularis for loop diuretics. As a previous study suggested initial lesions of stria vascularis, an attempt of comparison and of chronological study was made between the organ of Corti and stria vascularis lesions by kanamycin intoxication. The method was devised by J. M. ARAN, with electrophysiological control. We failed to find in the stria vascularis a radial or longitudinal pattern of lesions. We could not discern a chronological injury between the organ of Corti and stria vascularis because both were damaged even in the less deafened animals. Nevertheless, two facts were clarified: hair cell lesions are lysosomial as for the kidney lesions, while stria vascularis lesions are mitochondrial, melanine granulations play a part in drug metabolism (increased number, secretory aspect) and deserve further study.     

豚鼠耳蜗血管纹边缘细胞钾离子通道特性的研究

目的 研究单个豚鼠耳蜗血管纹边缘细胞的基本电生理特性。方法 应用耳蜗血管纹组织块培养技术和全细胞膜片钳技术，观察单个培养的豚鼠血管纹边缘细胞在电压钳模式下的电流特性。结果 边缘细胞上记录到钾离子电流，该钾通道有以下特性：①具有电压依赖性；②通道的活动可被4-氨基吡啶（4-aminopyridine，4-AP）和四乙铵（tetraethylammonium，TEA）在相对高浓度下阻滞；③细胞外钙离子浓度的减少可导致该钾通道电流的降低。结论 豚鼠耳蜗血管纹边缘细胞钾离子通道电流包括钙离子依赖性钾电流、外向延迟整流钾电流和A型钾电流。     

Effect of steroids on increased permeability of blood vessels of the stria vascularis after auditory ossicle vibration by a drill in otologic surgery

OBJECTIVES: The vibration caused by drills used for middle ear surgery is considered one of the causes of postoperative sensorineural deafness. Seki et al reported that when drill-induced damage was created in the auditory ossicles of guinea pigs, permeability across the capillary vessels in the stria vascularis increased significantly with the duration of drill-induced vibration. The present study was undertaken to examine changes in permeability across the stria vascularis capillaries following vibration in experimental animals pretreated with steroids, with the goal of developing a method of preventing a vibration-induced increase in permeability across these capillaries. METHODS: After an intravenous dose of hydrocortisone and horseradish peroxidase, the auditory ossicles of guinea pigs were vibrated with a drill for 60 seconds. RESULTS: Intravenous steroid administration before vibration reduced the leakage of horseradish peroxidase from the stria vascularis capillaries after vibration. CONCLUSIONS: The findings suggested that steroids suppress the increase in permeability across the stria vascularis capillaries that results from drill-induced vibration.     

耳蜗血管纹组织块的缘细胞培养

目的 :建立豚鼠耳蜗血管纹 (SV)组织块缘细胞 (MCs)的培养方法 ,为进一步研究药物耳毒性及其作用机制奠定基础。方法 :2 6只豚鼠按SV培养时间随机分成 4组 :2 4h组 (n =8) ;72h组 (n =8) ;>72h组 (n =8) ;对照组 (新鲜SV固定组 ,n =2 )。显微解剖数段连同螺旋韧带的SV组织块 ,置于 5 %CO2 / 95 %空气的二氧化碳恒温 (37℃ )培养箱中进行培养 ,分别进行形态学和组织学观察。结果 :培养 2 4hSV组织块保持良好活性 ,其组织学结构与新鲜固定的SV结构无明显差异 ;培养 72hSV组织块与新鲜固定的SV在组织学结构方面有显著性差异 ,不能观察到正常的SV结构 ,组织结构松散 ,缘细胞从组织块离心性生长出来 ;从SV组织块培养出的缘细胞能在培养皿内存活 13d。结论 :采用组织块培养技术 ,成功地建立了豚鼠耳蜗SV组织块的缘细胞培养方法 ;培养 2 4h的SV组织块光镜下保持了良好活性和正常组织学结构 ,可用来进一步研究药物耳毒性及其作用机制。     

Calmodulin binding sites in the endolymphatic sac and stria vascularis of the human fetus and the guinea pig.

Calmodulin (CaM) binding sites, which are available and unoccupied by CaM at the time of fixation, were analyzed in the guinea pig endolymphatic sac and stria vascularis using CaM conjugated tetramethyl rhodamine isothiocyanate. The endolymphatic sac epithelium and stria vascularis both showed strong fluorescence. Weak CaM binding sites were seen in the endolymphatic sac in the 12-week-old human fetus. In contrast, the fluorescent reaction in the stria vascularis and endolymphatic sac in 14- and 15-week-old human fetuses was strong. These results suggest that CaM may play an important role for the maintenance of ionic stability in the endolymph.