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A comparison of the cholinergic properties of the leopard frog vestibular organs 豹蛙前庭器官胆碱能特性的比较
J. C. Holt, P. Guth, P. Perin, Charles A. Norris
Previous data from this group indicated that the main cholinergic effect on the afferent resting firing rate of the posterior canal is an atropine-sensitive, strychnine-resistant facilitation, while the main effect on the saccule is a strychnine-sensitive inhibition. In the present research we compared the effect of acetylcholine (ACh) on the afferent whole-nerve resting discharge of all vestibular organs of the frog. All three semicircular canals, utricle and lagena responded to ACh and carbachol (CCh) perfusion with an increase in resting discharge, while the saccular afferent discharge was mainly inhibited. The perfusion with 10 μ M physostigmine potentiated both facilitatory and inhibitory responses to ACh in all organs tested. On the other hand, CCh was much more potent than ACh in producing the facilitatory response but not the inhibitory response. As already observed in the posterior canal, the facilitation observed in the other vestibular organs was reversibly blocked by 1 or 10 μ M atropine but not by 1 μ M strychnine. In addition, the irreversible non-selective muscarinic antagonist propylbenzilylcholine mustard irreversibly abolished the facilitatory response but not the inhibitory one, while 50 μ M tetraethylammonium blocked the inhibitory response without affecting the facilitatory response or producing inhibition by itself. These results suggest that the predominant cholinergic response on the resting discharge is muscarinic in all vestibular organs of the frog except the saccule.
本组前期数据表明,胆碱能对后管传入静息放电率的主要影响是阿托品敏感、士的宁抗性促进作用,而对囊的主要影响是士的宁敏感抑制作用。本研究比较了乙酰胆碱(ACh)对青蛙前庭各器官传入全神经静息放电的影响。3条半规管、胞室和筋膜对乙酰胆碱和碳水化合物(CCh)灌注均有反应,静息放电增加,而囊状传入放电主要受到抑制。灌注10 μ M的豆蔻碱可增强各器官对乙酰胆碱的促进和抑制反应。另一方面,CCh在产生促进反应方面比ACh更有效,而不是抑制反应。如前所述,1或10 μ M阿托品可逆转其他前庭器官的促进作用,而1 μ M士的宁则不能。不可逆非选择性毒蕈碱拮抗剂丙基苄基胆碱芥菜不可逆地消除了促进反应,但不消除抑制反应,而50 μ M四乙基铵阻断了抑制反应,但不影响促进反应或自身产生抑制作用。这些结果表明,除囊外,蛙的所有前庭器官在静息放电时的主要胆碱能反应是毒蕈碱。
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引用次数: 8
Novel calcium sensor in the outer hair cells: quantitation of oncomodulin 外毛细胞中的新型钙传感器:肿瘤调节素的定量
I. Thalmann, M. Henzl, R. Thalmann
The parvalbumins are small vertebrate-specific Ca 2+ -binding proteins. The mammalian α -parvalbumin is expressed in diverse cell types, including the inner hair cell, where it is proposed to serve as a cytosolic Ca 2+ buffer. By contrast, the mammalian β -parvalbumin (oncomodulin) has a highly restricted distribution and harbors a 'Ca 2+ -specific' site — properties consistent with Ca 2+ -dependent regulatory behavior. The widespread perception of oncomodulin as an oncofetal protein was recently shattered by our discovery that the protein is present in the organ of Corti. Immunohistochemical data for the rat, mouse and gerbil indicate that expression is in fact confined to the outer hair cell. We have sought to confirm and extend this work through biochemical quantitation studies on organ of Corti from rat, mouse, guinea pig and chinchilla. We find that the protein is expressed at comparably high levels (approximately 0.5 mM) in the outer hair cells in all four species. This finding contrasts sharply with the expression pattern in prenatal and neoplastic settings. We have also extended the immunohistochemical analysis to the guinea pig and performed preliminary analysis of oncomodulin mRNA levels by in situ hydridization. On the basis of our data, we suggest that the outer hair cell is the sole site of physiologically relevant oncomodulin expression. Furthermore, we propose that the protein functions in the efferent control of the electromotile response.
细小蛋白是脊椎动物特有的ca2 +结合蛋白。哺乳动物α -小白蛋白在多种细胞类型中表达,包括内毛细胞,它被认为是细胞质ca2 +缓冲液。相比之下,哺乳动物β -小白蛋白(肿瘤调节蛋白)具有高度受限的分布,并且具有“ca2 +特异性”的位点-与ca2 +依赖的调节行为一致。人们普遍认为肿瘤调节蛋白是一种癌胎蛋白,最近我们发现这种蛋白存在于Corti器官中,这一发现打破了人们对这种蛋白的普遍认识。大鼠、小鼠和沙鼠的免疫组织化学数据表明,表达实际上仅限于外毛细胞。我们试图通过对大鼠、小鼠、豚鼠和栗鼠Corti器官的生化定量研究来证实和扩展这一工作。我们发现,在所有四个物种的外毛细胞中,该蛋白的表达水平相当高(约0.5 mM)。这一发现与产前和肿瘤环境中的表达模式形成鲜明对比。我们还将免疫组织化学分析扩展到豚鼠,并通过原位氢化对肿瘤调节素mRNA水平进行了初步分析。根据我们的数据,我们认为外毛细胞是生理上相关的肿瘤调节素表达的唯一部位。此外,我们提出该蛋白在电运动反应的传出控制中起作用。
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引用次数: 7
Transformation of pressure- and heat-induced discharges of feline cutaneous C-fiber mechano-heat-sensitive units by lidocaine and N-propylajmaline 利多卡因和n -丙基马林对猫皮肤c -纤维机械热敏单元压力和热诱导放电的影响
Revenko, Ermishkin, Borovikov, Borovikova
Subcutaneous lidocaine (0.1%) and N-propylajmaline (NPA, 0.01%), inhibited pressure- and heat-evoked discharges in feline cutaneous C-fiber mechano-heat-sensitive (CMH) units. Initial suppression of discharges was followed by the period of resting and use-dependent (UD) inhibition, evaluated with double thermal (50 °C, 10 s) or triple mechanical (0.5 N/mm2, 3 s) stimuli, repeatedly applied after prolong pauses. Resting inhibition of pressure responses decreased the maximum discharge rate (MDR) evoked by the first stimulus in a train from the control value of 10.3 ± 1.5 Hz (n = 15, SEM) to 4.1± 1.0 Hz (n = 7) for lidocaine and to 6.3± 1.0 Hz (n = 15) for NPA. UD inhibition further decreased MDR evoked by the second and the third stimulus in a train: for lidocaine to 1.0 ± 0.7 Hz (n = 7) and 0.2 ± 0.1 Hz (n = 7); for NPA to 2.1 ± 0.8 Hz (n = 15) and 0.2 ± 0.1 Hz (n = 15). Resting inhibition of heat discharges decreased MDR evoked by the first stimulus from the control value of 6.0 ± 1.0 Hz (n = 28) to 2.4 ± 0.5 Hz (n = 8) for lidocaine and to 2.8 ± 0.3 Hz (n = 20) for NPA. UD inhibition further decreased MDR evoked by the second stimulus: in percentage to the first one, MDR was 57±14% (n = 8) for lidocaine and 56 ± 7% (n = 20) for NPA. Lidocaine and NPA transformed discharges evoked by a ramp heat stimulus, maintaining low frequencies (MDR < 2-3 Hz) and eliminating the higher ones. Tertiary amine lidocaine, but not quaternary NPA, elevated the thermal threshold by 2.8± 0.9 °C (n = 24, P < 0.01). UD inhibition of CMH unit termination indicates that it contains ionic channels with slow kinetics, thought to be tetrodotoxin-resistant sodium channels. Such UD inhibition allows us to explain the analgesic action of local anesthetics in low (subblocking) concentrations and propose local analgesia, which cuts the nociceptive signals off, but maintains the non-nociceptive ones.
皮下利多卡因(0.1%)和n -丙基马林(NPA, 0.01%)抑制猫皮肤c -纤维机械热敏(CMH)单元的压力和热诱发放电。最初的放电抑制之后是休息和使用依赖(UD)抑制,通过双热(50°C, 10 s)或三重机械(0.5 N/mm2, 3 s)刺激进行评估,在延长暂停后反复施加。静息抑制压力反应可使第一刺激引起的最大放电率(MDR)从对照值10.3±1.5 Hz (n = 15, SEM)降至利多卡因组的4.1±1.0 Hz (n = 7)和NPA组的6.3±1.0 Hz (n = 15)。UD抑制进一步降低了第二和第三种刺激引起的多药耐药:利多卡因为1.0±0.7 Hz (n = 7)和0.2±0.1 Hz (n = 7);为2.1±0.8 Hz (n = 15)和0.2±0.1 Hz (n = 15)。静息抑制热放电使第一刺激引起的MDR从利多卡因的6.0±1.0 Hz (n = 28)降低到2.4±0.5 Hz (n = 8), NPA的2.8±0.3 Hz (n = 20)。UD抑制进一步降低了第二种刺激引起的MDR:利多卡因的MDR为57±14% (n = 8), NPA的MDR为56±7% (n = 20)。利多卡因和NPA改变斜坡热刺激引起的放电,维持低频(MDR < 2 ~ 3hz),消除高频。叔胺利多卡因使大鼠热阈值升高2.8±0.9℃(n = 24, P < 0.01)。UD抑制CMH单元终止表明其含有缓慢动力学的离子通道,被认为是抗河豚毒素的钠通道。这种UD抑制使我们能够解释局部麻醉剂在低浓度(亚阻断)下的镇痛作用,并提出局部镇痛,它切断了伤害性信号,但保持了非伤害性信号。
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引用次数: 2
An intact peripheral nerve preparation for examining the central actions of single kinaesthetic afferent fibres arising in the wrist joint of the cat 完整的外周神经准备,用于检查猫手腕关节产生的单个动觉传入纤维的中枢作用
P. D. Mackie, Rowe, Coleman, Zhang
Intraneural microstimulation of single tactile or kinaesthetic afferent fibres arising in the hand of conscious human subjects has revealed marked differences among the different classes in their capacity to generate a perceptual response. In order to test the hypothesis that these different capacities might reflect a differential security in the transmission of singnals across synaptic junctions in the dorsal column nuclei or other levels of the somatosensory pathway we have previously developed a paired, simultaneous recording paradigm in the cat to analyze transmission characteristics within the dorsal column nuclei for single identified tactile and muscle sensory fibres of the forearm. These studies have depended upon the use of a fine peripheral nerve or nerve fascicle preparation in which it is possible to monitor the activity of single sensory fibres while the nerve remains in continuity with the central nervous system. Although we have previously described a preparation that allows the activity of single joint afferent fibres from the hindlimb to be monitored in the intact medial articular nerve, these hindlimb kinaesthetic afferents fail to project directly to the dorsal column nuclei. In the present study we report a forearm nerve preparation in the cat that permits the simultaneous recording of activity from individual wrist joint afferent fibres and their target neurones of the dorsal column nuclei. When this nerve is freed from nearby tissue over a distance of 2-4 cm and left in continuity, it is possible with a silver hook electrode to monitor the impulse activity of each group II joint afferent fibre with an excellent signal-to-noise ratio. The preparation should prove ideal for examining the central actions and security of transmission across the dorsal column nuclei for single, identified joint afferent fibres of the forearm.
对有意识的人的手部产生的单个触觉或动觉传入纤维进行神经内微刺激,揭示了不同类别的人在产生知觉反应的能力上的显著差异。为了验证这一假设,即这些不同的能力可能反映了信号在通过背柱核突触连接或其他水平的体感通路传递时的不同安全性,我们之前在猫身上开发了一种成对的同时记录范式,以分析前臂单个识别的触觉和肌肉感觉纤维在背柱核内的传递特征。这些研究依赖于使用精细的周围神经或神经束准备物,这样可以在神经与中枢神经系统保持连续性的同时监测单个感觉纤维的活动。尽管我们之前已经描述了一种制备方法,可以在完整的内侧关节神经中监测来自后肢的单个关节传入纤维的活动,但这些后肢动觉传入神经不能直接投射到背柱核。在本研究中,我们报告了猫的前臂神经准备,允许同时记录单个手腕关节传入纤维及其背柱核的目标神经元的活动。当该神经与附近组织分离2-4厘米并保持连续性时,可以用银钩电极监测每组II关节传入纤维的脉冲活动,并具有良好的信噪比。这种制备方法对于检查前臂单个、可识别的关节传入纤维的中枢作用和通过背柱核传递的安全性是理想的。
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引用次数: 5
The frequency dependence of receptor potentials in hair cells of the mouse utricle 小鼠小囊毛细胞受体电位的频率依赖性
A. Rüsch, J. R. Holt, R. Eatock, Melissa A. Vollrath
The mechanoelectrical transduction currents of hair cells in the mouse utricle adapt at varying rates to step deflections of the hair bundles. We consider contributions of this adaptation process and of input resistance and membrane capacitance to the frequency dependence of the receptor potential. Whole-cell recordings of transduction current and receptor potential were made from hair cells in the excised epithelium of the mouse utricle. Hair bundles were deflected by a fluid jet with step and sinusoidal waveforms. In type II cells, the receptor potential was a bandpass function of stimulus frequency. The adaptation rate of the transduction current, measured in response to step bundle deflections, accounted for much of the roll-off in the receptor potential at low frequencies of sinusoidal deflections. Corner frequencies predicted from the adaptation time course varied from 2 to 60 Hz. Voltage-gated conductances also contributed. Roll-off of the receptor potential at the high-frequency end may largely reflect input resistance and capacitance. Corner frequencies predicted by estimated membrane time constants varied from 30 to 150 Hz. In type I cells, slower or no adaptation and shorter membrane time constants predict larger response bandwidths. Frequency tuning in vivo will reflect other factors, including the mechanical response of the otolith and otolithic membrane to head movements.
小鼠胞内毛细胞的机电转导电流以不同的速率适应毛束的阶跃偏转。我们考虑了这种适应过程以及输入电阻和膜电容对受体电位频率依赖性的贡献。在小鼠胞室切除上皮毛细胞中记录了传导电流和受体电位的全细胞记录。毛束被具有阶跃和正弦波形的流体射流偏转。在II型细胞中,受体电位是刺激频率的带通函数。根据阶跃束偏转测量的转导电流的适应率,在低频正弦偏转下占受体电位滚转的大部分。根据自适应时间过程预测的角频率在2 ~ 60hz之间变化。电压门控电导也起了作用。高频端受体电位的滚降在很大程度上反映了输入电阻和电容。估计膜时间常数预测的角频率在30 ~ 150hz之间变化。在I型细胞中,较慢或不适应和较短的膜时间常数预示着较大的响应带宽。体内的频率调整将反映其他因素,包括耳石和耳石膜对头部运动的机械响应。
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引用次数: 8
Amiloride high- and low-sensitivity, as well as insensitive sites in the blowfly: implications for sugar, water and anion taste reception mechanisms 阿米洛利在苍蝇体内的高、低敏感性和不敏感部位:对糖、水和阴离子味觉接受机制的影响
Majone, A. L. P.Solari, Crnjar
Among chemoreceptor cells of the blowfly ( Protophormia terraenovae , the so-called 'sugar' and 'water' cells respond to bovine serum albumin (BSA) and L -alanine (the C-terminal amino acid of the BSA molecule) with the highest spike frequency. In both cell types the response to either BSA or L -alanine was unaffected by 0.1 mM amiloride. However, 0.5 mM amiloride decreased the response of the 'water' cell to both BSA and L-alanine significantly, and similarly inhibited the response of the 'sugar' cell to L -alanine, but in contrast to the 'water' cell, 0.5 mM amiloride did not affect the 'sugar' cell response to BSA. The 'sugar' cell responses to both BSA and L-alanine were unaffected by the putative cell second messengers, cAMP and cGMP. The so-called 'anion' cell unexpectedly gave responses to BSA and L-alanine that were enhanced by amiloride at both concentrations. However, like the 'sugar' cell, the 'anion' cell was also unaffected by cAMP and cGMP.We conclude that the 'sugar' cell must have at least three types of receptor sites: the previously described 'F' (amiloride sensitive) and the 'P' (amiloride insensitive) sites, and a low-sensitivity 'T' site that mediates, at least in part, the response to BSA and L -alanine. The effects of amiloride on the responses of the 'water' and 'anion' cells are more difficult to interpret because fundamental information on their chemoreception mechanisms is still lacking.
在苍蝇的化学感受器细胞中,所谓的“糖”和“水”细胞对牛血清白蛋白(BSA)和L -丙氨酸(BSA分子的c端氨基酸)的反应频率最高。在这两种细胞类型中,对BSA或L -丙氨酸的反应均不受0.1 mM阿米洛利的影响。然而,0.5 mM amiloride显著降低了“水”细胞对BSA和L-丙氨酸的反应,同样抑制了“糖”细胞对L-丙氨酸的反应,但与“水”细胞相比,0.5 mM amiloride不影响“糖”细胞对BSA的反应。糖细胞对BSA和l -丙氨酸的反应不受假定的细胞第二信使cAMP和cGMP的影响。所谓的“阴离子”细胞意外地对两种浓度的阿米洛利都增强的BSA和l -丙氨酸产生反应。然而,像糖细胞一样,阴离子细胞也不受cAMP和cGMP的影响。我们得出结论,“糖”细胞必须至少有三种类型的受体位点:先前描述的“F”(阿米洛利敏感)和“P”(阿米洛利不敏感)位点,以及至少部分介导对BSA和L -丙氨酸反应的低敏感性“T”位点。阿米洛利对“水”和“阴离子”细胞反应的影响更难解释,因为它们的化学接受机制的基本信息仍然缺乏。
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引用次数: 7
Chicken (Gallus domesticus) inner ear afferents. 鸡(家鸡)内耳传入。
Hirota Hara, Xiangdong Chen, Jershonda F. Hartsfield, J. Hara, D. Martin, C. Fermin
Neurons from the vestibular (VG) and the statoacoustic (SAG) ganglion of the chick (Gallus domesticus) were evaluated histologically and morphometrically. Embryos at stages 34 (E8 days), 39 (E13 days) and 44 (E18 days) were sacrificed and temporal bones microdissected. Specimens were embedded in JB-4 methacrylate plastic, and stained with a mixture of 0.2% toluidine blue (TB) and 0.1% basic Fuschin in 25% ethanol or with a mixture of 2% TB and 1% paraphenylenediamine (PDA) for axon and myelin measurement study. Images of the VIIIth nerve were produced by a V150 (R) color imaging system and the contour of 200-300 neuronal bodies (perikarya) was traced directly on a video screen with a mouse in real time. The cross-sectional area of VG perikarya was 67.29 micrometers2 at stage 34 (E8), 128.46 micrometers2 at stage 39 (E13) and 275.85 micrometers2 at stage 44 (E18). The cross-sectional area of SAG perikarya was 62.44 micrometers2 at stage 34 (E8), 102.05 micrometers2 at stage 39 (E13) and 165.02 micrometers2 at stage 44 (E18). A significant cross-sectional area increase of the VG perikarya between stage 39 (E13) and stage 44 (E18) was determined. We randomly measured the cross-sectional area of myelin and axoplasm of hatchling afferent nerves, and found a correspondence between axoplasmic and myelin cross-sectional area in the utricular, saccular and semicircular canal nerve branches of the nerve. The results suggest that the period between stage 34 (E8) and 39 (E13) is a critical period for afferent neuronal development. Physiological and behavioral vestibular properties of developing and maturing hatchlings may change accordingly. The results compliment previous work by other investigators and provide valuable anatomical measures useful to correlate physiological data obtained from stimulation of the whole nerve or its parts.
对鸡(Gallus domesticus)前庭神经节(VG)和静声神经节(SAG)的神经元进行了组织学和形态学评价。处死第34期(E8 d)、第39期(E13 d)和第44期(E18 d)胚胎,显微解剖颞骨。将标本包埋在j -4甲基丙烯酸酯塑料中,用0.2%甲苯胺蓝(TB)和0.1%碱性Fuschin在25%乙醇中或2% TB和1%对苯二胺(PDA)的混合物染色,用于轴突和髓鞘的测量研究。采用V150 (R)彩色成像系统对第viii神经进行成像,用鼠标直接在视频屏幕上实时追踪200-300个神经元体(核周)的轮廓。第34期(E8)、第39期(E13)和第44期(E18)的横截面积分别为67.29微米、128.46微米和275.85微米。第34期(E8)、第39期(E13)和第44期(E18)核周截面积分别为62.44、102.05和165.02微米。在第39期(E13)和第44期(E18)之间,VG核周的横截面积显著增加。我们随机测量了幼雏传入神经的髓磷脂和轴质横截面积,发现在神经的室、囊和半规管神经分支中,轴浆和髓磷脂横截面积具有对应关系。提示第34期(E8)至第39期(E13)是传入神经元发育的关键时期。发育和成熟的幼雏前庭系统的生理和行为特性可能会发生相应的变化。该结果补充了其他研究者先前的工作,并提供了有价值的解剖学测量,有助于将整个神经或其部分刺激获得的生理数据联系起来。
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引用次数: 3
The effect of tetraethylammonium and 4-aminopyridine on mechano- and electrosensitive channels of the Pacinian corpuscle 四乙基铵和4-氨基吡啶对太平洋小体机械和电敏通道的影响
Krylov, Dick, Akoev
The action of tetraethylammonium (TEA) (20 mM) and 4-aminopyridine (4-AP) (5 mM) on mechano- and electrically excitable membranes of the Pacinian corpuscles was studied by means of the air gap technique in constantly perfused preparations. Extracellular recordings of receptor potential have shown that application of TEA causes 1.5-fold prolongation of the receptor potential and a decrease of its amplitude by 40%. 4-AP has no effect on the mechanosensitive membrane. The difference in the blockers effect testifies that repolarization of the receptor potential is not regulated by the voltage-dependent potassium channels. The blockers considered are likely effect on the calcium-dependent potassium currents activated by Ca 2+ ions passing through the non-selective channels opening in response to mechanical stimulus. TEA and 4-AP induce a 2- to 3-fold increase of the action potential duration in the electrically excitable membrane of the Ranvier node. Computations based on the Dodge model help reveal that inhibition of the voltage-dependent potassium channels accounts for this increase.
采用气隙法研究了四乙基铵(TEA) (20 mM)和4-氨基吡啶(4-AP) (5 mM)对太平洋小体机械和电兴奋膜的作用。受体电位的细胞外记录表明,TEA的应用导致受体电位延长1.5倍,其振幅下降40%。4-AP对机械敏感膜无影响。阻滞剂效应的差异证明受体电位的复极化不受电压依赖性钾通道的调节。所考虑的阻滞剂可能影响钙依赖性钾电流,钙离子通过机械刺激下开放的非选择性通道激活钾电流。TEA和4-AP诱导Ranvier结电兴奋膜的动作电位持续时间增加2- 3倍。基于道奇模型的计算有助于揭示电压依赖性钾通道的抑制作用导致了这种增加。
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引用次数: 0
Glutamic acid decarboxylase in the rat ampullary cristae: immunocytochemical and immunoblotting studies 大鼠壶腹嵴谷氨酸脱羧酶:免疫细胞化学和免疫印迹研究
G. Meza, Jang-Yen Wu, H. Zepeda, C. José
In order to investigate a putative neurotransmitter function of γ-aminobutyric acid (GABA) in the rat vestibule we chose to study the cellular localization and properties of glutamic acid decarboxylase (GAD), the rate-limiting enzyme for the synthesis of GABA, in the rat ampullary cristae, with respective immunocytochemical and immunoblotting techniques, using an extensively characterized rabbit anti-GAD serum. With these procedures we found a GAD-like immunopositivity in the sensory cells and in fibers of the crista ampullaris stroma. In ampullary cristae homogenates, a GAD composed of at least two subunits (53 and 67 kDa which were present also in rat brain and cerebellum homogenates) was encountered. These findings suggest that GAD present in the rat vestibule is homologous to the brain enzyme and it has the appropriate localization to synthesize GABA to be used as a neurotransmitter in the rat vestibule.
为了研究γ-氨基丁酸(GABA)在大鼠前庭中的神经递质功能,我们选择利用免疫细胞化学和免疫印迹技术,研究大鼠壶腹嵴中合成GABA的限速酶谷氨酸脱羧酶(GAD)的细胞定位和性质,使用广泛表征的兔抗GAD血清。通过这些程序,我们发现壶腹嵴间质感觉细胞和纤维中的gad样免疫阳性。在壶腹嵴匀浆中,发现了至少由两个亚基(53和67 kDa)组成的GAD,它们也存在于大鼠脑和小脑匀浆中。这些发现表明,GAD存在于大鼠前庭中,与脑酶同源,并且在大鼠前庭中具有合适的定位,可以合成GABA作为神经递质。
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引用次数: 1
Hair cells of a congenitally hearing impaired canary have abnormal distribution of filamentous proteins 先天性听力受损金丝雀毛细胞中丝状蛋白分布异常
T. Park, P. Weisleder, Ying Lu
Previous investigations into the anatomy of the inner ear of Belgian Waterslager canaries (BWC) have demonstrated myriad malformations associated with dysgenesis of the pars inferior of the otocyst. These malformations are associated with a congenital hearing impairment primarily affecting mid to high frequencies. Specific hair cell abnormalities include irregular cell boundaries, deformed cuticular plate, and multiple and deformed stereocilia tufts. This constellation of structural deformities lead us to hypothesize that the content of non-muscle filamentous protein in BWC hair cells need be abnormal. As a first step in exploring this hypothesis, we have used immunocytochemical techniques to assess the presence and distribution of the major components of the actin microfilament system, i.e. actin, tropomyosin and α -actinin, at the apical end of hair cells from the BWC basilar papilla. Our results show qualitative differences between control and BWC hair cells. First, label of the BWC cuticular plate was scattered and ill-matched with that of control birds for each of the three proteins examined. Second, labeling of BWC cilia revealed abnormal distribution of the assessed proteins. Specifically, anti-actin and anti- α -actinin antibodies were clustered over the cilia, while immunostaining against tropomyosin was not seen. These findings support a link between the proteins associated with cochlear hair cell structural integrity and the dysmorphologies associated with the congenital hearing impairment identified in the BWC model.
先前对比利时水斯莱格金丝雀(BWC)内耳解剖结构的调查已经证明了与耳囊肿下部发育不良相关的无数畸形。这些畸形与先天性听力障碍有关,主要影响中高频。特定的毛细胞异常包括细胞边界不规则,角质层变形,多发和变形的立体纤毛簇。这些结构畸形使我们推测BWC毛细胞中非肌丝状蛋白的含量可能异常。作为探索这一假设的第一步,我们使用免疫细胞化学技术评估了肌动蛋白微丝系统的主要成分,即肌动蛋白、原肌球蛋白和α -肌动蛋白在BWC基底乳头毛细胞顶端的存在和分布。我们的结果显示了对照组和BWC毛细胞之间的质量差异。首先,BWC角质层板上的三种蛋白质的标签分散,与对照鸟的标签不匹配。其次,BWC纤毛的标记显示评估蛋白的异常分布。具体来说,抗肌动蛋白和抗α -肌动蛋白抗体聚集在纤毛上,而未见针对原肌球蛋白的免疫染色。这些发现支持与耳蜗毛细胞结构完整性相关的蛋白质与BWC模型中发现的先天性听力障碍相关的畸形之间的联系。
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Primary sensory neuron : the international interdisciplinary journal reporting basic and clinical research on sensory receptors and primary afferent neurons
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