定量观察卡铂引起的灰鼠耳蜗毛细胞和神经纤维的早期损害过程

目的：定量观察和比较灰鼠耳蜗毛细胞和缰孔内神经纤维数量在卡铂耳中毒早期损害过程中的变化。方法：在耳蜗基底膜铺片的基础上制备耳蜗毛细胞图；从骨性螺旋板切片上对缰孔内的神经纤维计数。结果：缰孔内的神经纤维数量在注射卡铂后24h内明显减少。而内毛细胞的缺失是发生在注射上学铂后72h。结论：在卡铂耳中毒早期，缰孔内神经纤维的破坏早于内毛细胞的缺失。     

灰鼠耳蜗毛细胞脱氢酶活性在卡铂耳中毒中的早期变化

采用组织化学技术观察了灰鼠耳蜗行细胞中脱氢酶卡铂耳中毒早期的活性变化。发现卡狂可选择性地抑制内毛细胞中的脱氢酶活性，并且这种影响是从底回开始向顶回发展。提示在内毛细胞变生之前，卡铂可通过抑制内毛细胞的脱氢酶干扰其能量的代谢。     

卡铂对灰鼠螺旋神经节的早期损害

实验中观察了卡铂(50mg/kg)对灰鼠耳蜗螺旋神经节细胞的急性损害.注药后6小时即可发现在Ⅰ型神经元胞体和神经纤维中有空泡形成.注药后1天可见Ⅰ型神经元内充满大空泡并开始发生坏死.其结果表明卡铂产生对螺旋神经节的毒害相当迅速.提示耳蜗传入神经系统可能是卡铂的第一个靶组织.     

卡铂引起的灰鼠耳蜗内毛细胞缺损模式

目的:研究卡铂损害灰鼠耳蜗内毛细胞(IHCs)的模式,即IHCs何时出现缺损、给药后不同时间基底膜上IHCs缺损的范围、基底膜上IHCs最大缺损部位以及致绝大部分IHCs缺损所需时间等。方法:对成年灰鼠采用一次性卡铂腹腔注射(100mg/kg),给药后不同时间处死动物,常规制备耳蜗铺片和耳蜗图,以定量观察灰鼠耳蜗毛细胞的缺损情况。结果:注射卡铂后从24h～3个月,灰鼠的外毛细胞基本无缺损。IHCs损伤模式的特征:①注射卡铂后24h耳蜗毛细胞完整无损,IHCs从注药后第3天开始出现缺损;②从注射卡铂后3d到3个月期间,IHCs的最大缺损率均出现在第1回和第2回交界处;③IHCs缺损从第1回和第2回交界处开始,范围逐渐向底回和顶回扩展,与顶回相比,底回的IHCs缺损出现早且更严重;④至注射后3个月基底膜上IHCs的缺损呈平坦型。结论:100mg/kg卡铂腹腔注射对各回IHCs的破坏是不均等的,对卡铂的敏感度不一致可能造成了各回IHCs缺损在时间上的差异。     

P物质抗体对豚鼠耳蜗核下丘核团内听觉诱发电位调谐曲线的影响

探索Ｐ物质在听觉脑干中枢中对声信号的频率分析作用。方法采用短音刺激,短纯音前掩蔽法和豚鼠耳蜗核、正丘、核团内电极,记录ＣＮ及ＩＣ核团内听觉诱发电位。观察核团内注射微量Ｐ物质抗体或对照注射等量兔血清后ＣＮ和ＩＣ核团内听觉诱发电位调谐曲线的变化。     

卡铂及其耳毒性

卡铂作为一种铂制剂广泛应用于临床抗肿瘤的化学治疗，其毒副作用主要包括耳毒性，肾毒性，神经毒性．和抑制骨髓造血系统等等。卡铂可以选择性破坏灰鼠的耳蜗内毛细胞和前庭I型毛细胞以及与之相联系的传入神经元。在卡铂引起的灰鼠单纯内毛细胞部分损害动物模型，动物具有正常的耳声发射和微音器电位以及听神经动作电位阈值．但总和电位和听神经动作电位的振幅却明显减小。在卡铂造成60％-90％的内毛细胞缺损的动物模型中，灰鼠在安静条件下对纯音的听觉条件反射阈可以保持正常，但在宽带或窄带噪声的掩蔽作用下，其纯音听觉反射阈值却明显提高，同时对听觉瞬时清晰度的辨别能力也明显降低。卡铂引起的内毛细胞和前庭I型毛细胞及其传入数据元的破坏过程往往伴随着钙激活蛋白酶的活动和Caspase-8的启动以及p53的过度调控从而引起细胞凋亡，同时可见琥珀酸脱氢酶，钠钾ATP酶，碱性磷酸酶和葡萄糖-6-磷酸酶活性的减弱以及钙激活ATP酶活性的增强。无论在灰鼠的活体动物实验还是离体培养条件下．卡铂都对灰鼠耳蜗内毛细胞和前庭I型毛细胞具有特殊的选择性破坏作用。卡铂对灰鼠听觉系统的损害除了导致内毛细胞和I型螺旋神经节凋亡之外。还会破坏耳蜗核神经元。但对下丘和听皮层的神经元却不造成致命性损坏。在耳蜗内毛细胞和I型螺旋神经节以及耳蜗核神经元被卡铂破坏之后．下丘和听皮层神经元对微弱的输入声刺激信号却表现出超乎寻常的敏感性提高。提示中枢神经元可能通过降低其某些抑制性神经递质的释放使神经元的活动代偿性增强．亦可能通过加强兴奋性神经递质的释放．或者通过神经元新长出的轴突和末梢而接受到更多的刺激信号，从而弥补了因周边听觉输入信号减弱而引起的听功能障碍，因此，卡铂耳中毒灰鼠模型还可成为一个研究听觉中枢重组现象或听神经病的理想实验动物模型。     

灰鼠下丘近场记录未观察到掩蔽级差(Masking Level Difference)现象

目的 以往的实验中采用近场电极记录方法发现了听皮层电位的掩蔽级差(Masking level difference,MLD)样改变,本实验采用同样技术观察下丘的MLD是否存在。方法6只健康成年灰鼠,手术植入右侧下丘电极,术后恢复期7-10天,然后在无麻醉状态下进行下丘电位的测试。短纯音条件的测试(500 Hz短纯音)共设:双耳同相纯青(b0),双耳反相纯音(bπ),左耳纯音(le)和右耳纯音(re)四个条件,用以观察纯音本身参数对实验的影响。MLD条件的测试(500 Hz短纯音+白噪声)共设:双耳同相噪声反相纯音(n0sπ),双耳同相噪声同相纯音(n0s0),双耳同相噪声左耳纯音(n0sl),左耳噪声左耳纯音(nlsl),双耳同相噪声右耳纯音(n0sr)和右耳噪声右耳纯音(nrsr)六个条件。结果 纯音条件组的右侧声刺激(re)得到较弱的诱发电位。MLD条件组中下丘电位的噪声掩蔽阈,潜伏期和振幅的分析未发现各组数据间有明显差异。结论 采用近电场电极记录方法不能发现下丘的MLD样活动,但能发现皮层的MLD样活动。研究结果支持MLD完成于脑干以上水平的观点。     

灰鼠耳蜗毛细胞脱氢酶活性在卡铂耳中毒时的早期变化

采用组织化学技术观察了灰鼠耳蜗毛细胞中脱氢酶在卡铂耳中毒早期的活性变化.发现卡铂可选择性地抑制内毛细胞中的脱氢酶活性.并且这种影响是从底回开始向顶回发展.提示在内毛细胞变性之前.卡铂可通过抑制内毛细胞的脱氢酶干扰其能量的代谢.     

Evidence for increased NADPH-diaphorase-positive neurons in the central auditory system of the aged rat

Conclusions: The age-related increase in the production of nitric oxide (NO) suggests that this increase was related to neuron aging. Additional studies may provide information regarding aging-related changes in the central auditory system. Objectives: Although NO has been associated with aging, it is unclear whether specific areas of the central auditory system are involved. We therefore assayed aging-related changes in NADPH-diaphorase (NADPH-d), a selective histochemical marker for NO, in the neurons of the central auditory system and other brain regions. Materials and methods: The numbers of NADPH-d-stained neurons and the area and staining density of cell bodies were examined in aged (24 months old) and younger (4 months old) Wistar rats. Results: The number of NADPH-d-positive neurons in the inferior colliculus was significantly increased in aged rats (p<0.05), whereas the area of NADPH-d-positive neurons in all areas did not differ significantly between aged and younger rats (p>0.05). The staining densities of NADPH-d-positive neurons in the inferior colliculus, the auditory cortex, and the visual cortex were significantly greater in aged compared with younger rats (p<0.05).     

等调制深度时间调制转换曲线评估豚鼠下丘和听皮层的时间分辨率

目的探讨以听觉系统对调幅信号响应幅值随调制频率的改变计算等调制深度时间调制转换函数（temporal modulation transfer function，TMTF）来客观评估听觉系统时间分辨率的可行性。方法豚鼠下丘和听皮层分别埋植慢性电极，记录正弦调幅纯音（载频为8kHz，调制深度固定为100％）诱发电位，反应幅值经快速傅立叶变换（fast Fourier transform，F丌）得出相对反应幅值，并以相对反应幅值和调制频率绘制出等调制深度TMTF。记录正弦调幅纯音每个调制频率的调制深度从100％至10％的诱发电位，得出与传统的调制深度阈值TMTF方法相当的等幅值TMTF，与等调制深度TMTF相比较，判断等调制深度TMTF方法的有效性。结果豚鼠下丘和听皮层的等调制深度TMTF与等幅值TMTF都分别表现为带通和低通特性；等调制深度TMTF的截止频率与等幅值TMTF的截止频率差异无统计学意义（P值均〉0．05）。豚鼠听皮层等调制深度TMTF的截止频率与传统的行为学结果基本一致。结论以100％调制深度的正弦调幅纯音诱发反应幅值与调制频率绘制等调制深度TMTF是一种有效的客观评估听觉系统时间分辨率的方法，其中豚鼠听皮层的等调制深度TMTF可用于行为学预测。     

Late onset of auditory deprivation does not affect brainstem auditory neuron soma size

Neuronal soma sizes from normal mice, mice with conductive losses from 45 days, and avocal mice subjected to sound deprivation from 45 days, were studied at 90 days, revealing no significant differences between groups for cochlear nuclear globular cells, or cells of medial nucleus of trapezoid body or inferior colliculus.     

Single-unit responses to cholinergic agents in the rat inferior colliculus

Single units were recorded from the inferior colliculi of adult male rats anesthetized with urethane. Units were driven with tonal stimuli, and changes in unit responses to the tones were monitored during iontophoretic application of cholinergic drugs. The cholinergic agonists acetylcholine and carbamylcholine potentiated responses in about 50% and suppressed responses in about 35% of units tested. Cholinergic antagonists typically produced effects when delivered alone. Both d-tubocurarine and atropine methyl nitrate excited over 80% of units tested, while mecamylamine and scopolamine inhibited the majority of tested units. Dihydro-beta-erythroidine was generally ineffective. Alpha-bungarotoxin was generally ineffective when delivered alone, but blocked agonist effects. Post-stimulus time histogram (PSTH) patterns, response-intensity functions and response areas were examined for changes during drug delivery. Cholinergic agents did not differentially affect either time periods within the PSTH or frequency bands of response areas, but were especially effective for those intensities producing larger response rates. Taken together with evidence from biochemical studies, our results suggest the presence of a functional cholinergic input into the inferior colliculus which acts to modulate acoustic processing.     

The effect of noise-induced sloping high-frequency hearing loss on the gap-response in the inferior colliculus and auditory cortex of guinea pigs

Gap detection has been used as an evaluation tool for temporal processing in subjects with sensorineural hearing loss (SNHL). However, the results from other reports are varied making it difficult to clearly define the impact of SNHL on the temporal processing ability of the auditory system. Specifically, we do not know if and how a high-frequency hearing loss impacts, presumably through off-channel interaction, the temporal processing in low-frequency channels where hearing sensitivity is virtually normal. In this experiment, gap-evoked responses in a low-frequency band (0.5–8 kHz) were recorded in the inferior colliculus (IC) and auditory cortex (AC) of guinea pigs through implanted electrodes, before and after a slopping high-frequency hearing loss, which was induced by over-stimulation using a 12-kHz-tone. The results showed that the gap thresholds in the low-frequency region increased gradually and became significantly higher 8 weeks after the induced high-frequency hearing loss. In addition, the response latency was slightly increased in the IC but this was not true for the AC. These results strongly indicate that a high-frequency hearing loss exerted an off-channel impact on temporal processing in the low-frequency region of the auditory system.     

Central auditory plasticity after carboplatin-induced unilateral inner ear damage in the chinchilla: Up-regulation of GAP-43 in the ventral cochlear nucleus

Inner ear damage may lead to structural changes in the central auditory system. In rat and chinchilla, cochlear ablation and noise trauma result in fiber growth and synaptogenesis in the ventral cochlear nucleus (VCN). In this study, we documented the relationship between carboplatin-induced hair cell degeneration and VCN plasticity in the chinchilla. Unilateral application of carboplatin (5 mg/ml) on the round window membrane resulted in massive hair cell loss. Outer hair cell degeneration showed a pronounced basal-to-apical gradient while inner hair cell loss was more equally distributed throughout the cochlea. Expression of the growth associated protein GAP-43, a well-established marker for synaptic plasticity, was up-regulated in the ipsilateral VCN at 15 and 31 days post-carboplatin, but not at 3 and 7 days. In contrast, the dorsal cochlear nucleus showed only little change. In VCN, the high-frequency area dorsally showed slightly yet significantly stronger GAP-43 up-regulation than the low-frequency area ventrally, possibly reflecting the high-to-low frequency gradient of hair cell degeneration. Synaptic modification or formation of new synapses may be a homeostatic process to re-adjust mismatched inputs from two ears. Alternatively, massive fiber growth may represent a deleterious process causing central hyperactivity that leads to loudness recruitment or tinnitus.     

The effect of gap-marker spectrum on gap-evoked auditory response from the inferior colliculus and auditory cortex of guinea pigs

The objective of this study is to verify the effects of gap marker spectrum on gap-evoked auditory responses. The gap-evoked potentials were recorded using electrodes implanted in the inferior colliculus (IC) and auditory cortex (AC) of guinea pigs. The gap markers were noise bursts in four frequency bands (500–8?000?Hz, 500–16?000?Hz, 500–32?000?Hz, and 16?000–32?000?Hz), and were tested at three sound levels. The onset response to the post-gap marker was measured to obtain the gap response threshold, and to establish input-output functions for latency and amplitude. Similar to previous behavioural studies, it was found that the gap-response threshold decreased with increasing marker bandwidth. This change was more significant at the cortical level in which the averaged gap-threshold decreased by approximately 2?ms with the bandwidth change. However, the gap threshold in the high frequency region (16?000–32?000?Hz) was comparable to that of the low frequency region (500–16?000?Hz). These results suggest that the total bandwidth of all auditory channels that are recruited determine the temporal resolution measured in gap-evoked potentials.

Sumario

El objetivo de este estudio es verificar los efectos del espectro de marcadores de brecha sobre las respuestas auditivas evocadas por una brecha. Se registraron los potenciales evocados por brecha utilizando electrodos implantados en el colículo inferior (IC) y la corteza auditiva (AC) de cobayos. Los marcadores de brecha fueron bursts de ruido en cuatro bandas de frecuencia (500–8000?Hz, 500–16?000?Hz, 500–32?000?Hz and 16?000–32?000?Hz), y se evaluaron a tres niveles sonoros. Se midió la respuesta de inicio del marcador post-brecha para obtener el umbral de respuesta de brecha, y establecer las funciones de ingreso-salida para latencia y amplitud. Al igual que en estudios conductuales previos, se encontró que el umbral de respuesta de brecha disminuyó con el incremento de los anchos de banda del marcador. Este cambio fue más significativo a nivel cortical en donde el umbral promediado de brecha disminuyó en aproximadamente 2 mseg con el cambio de ancho de banda. Sin embargo, el umbral de brecha en la región de alta frecuencia (16?000–32?000?Hz) fue comparable al umbral de la región de baja frecuencia (500–16?000?Hz). Estos resultados sugieren que el ancho de banda total de todos los canales auditivos reclutados determina la resolución temporal medida en los potenciales evocados por brecha.     

Projections from auditory cortex contact cells in the cochlear nucleus that project to the inferior colliculus

Anterograde and retrograde tracing techniques were combined to determine whether auditory cortical axons contact cells in the cochlear nucleus that project to the inferior colliculus. FluoroRuby or fluorescein dextran was injected into auditory cortex to label cortical axons by anterograde transport. Different fluorescent tracers (Fast Blue, FluoroGold, FluoroRuby or fluorescein dextran) were injected into one or both inferior colliculi to label cells in the cochlear nucleus. After 12-15 days, the brain was processed for fluorescence microscopy and the cochlear nuclei were examined for apparent contacts between cortical axons and retrogradely labeled cochlear nucleus cells. The results suggest that axons from the ipsilateral or contralateral cortex contact fusiform and giant cells in the dorsal cochlear nucleus and multipolar cells in the ventral cochlear nucleus that project directly to the inferior colliculus. The contacts occur on cell bodies and dendrites. The target cells in the cochlear nucleus include cells that project ipsilaterally, contralaterally or bilaterally to the inferior colliculus. The results suggest that auditory cortex is in a position to exert direct effects on the monaural pathways that ascend from the cochlear nucleus.     

Low frequency neurons in the lateral central nucleus of the cat inferior colliculus receive their input predominantly from the medial superior olive

When iontophoretic injections of horseradish peroxidase were made in the vicinity of inferior colliculus units in the cat responding to low sound frequencies, retrograde labelling occurred in the ipsilateral medial superior olive. Both bipolar and multipolar cells of the medial superior olive participated in this projection, and the focus of labelling shifted topographically within the olive as the injection site best frequency varied. These observations confirm previous anatomical findings and link them to the tonotopic organization of the central nucleus of the inferior colliculus. However, the fact that the medial superior olive alone provides between 50 and 98% of labelled cells in the brain stem projecting to this region of the central nucleus is an unexpected observation. This study gives further support to an hypothesis of 'core zones' within the central nucleus that receive preferentially input from specific brain stem auditory nuclei.     

Isolation and chemical analysis of neuron soma samples from the cat cochlear nucleus

Pieces of neuron somata were dissected from the cochlear nuclei of two cats and analyzed for activity of malic dehydrogenase, an important enzyme of oxidative metabolism. Based on location, size and shape the somata were identified as those of spherical bushy cells, octopus cells, and fusiform cells. Somata of cerebellar Purkinje cells were analyzed for comparison. The results suggest that the large cochlear nucleus neurons have malic dehydrogenase activities higher than those of most other neurons so far studied. Also, the octopus and fusiform somata had higher malic dehydrogenase activities than those of the spherical bushy cells.