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首页> 外文期刊>The European Journal of Neuroscience >Glutamate transporters in the guinea-pig cochlea: partial mRNA sequences, cellular expression and functional implications.
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Glutamate transporters in the guinea-pig cochlea: partial mRNA sequences, cellular expression and functional implications.

机译：豚鼠耳蜗中的谷氨酸转运蛋白：部分mRNA序列，细胞表达和功能含义。

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摘要

In the cochlea, glutamate plays a major role in synaptic transmission between the inner hair cell and the primary auditory neurons. Extracellular glutamate concentration must be regulated to prevent excitotoxicity. This regulation is mediated by excitatory amino acid transporters, membrane proteins that remove glutamate from the synaptic cleft. In this study, we investigated the distribution and activity of three excitatory amino acid transporters subtypes in the guinea-pig cochlea: glutamate aspartate transporter, glutamate transporter and excitatory amino acid carrier. A partial messenger ribonucleic acid sequence was determined for each of these transporters, by polymerase chain reaction with degenerate primers, using guinea-pig brain complementary deoxyribonucleic acid as the template. Primers specific for each transporter were then designed and used to screen a dissected organ of Corti complementary deoxyribonucleic acid library. The cellular distribution of each transporter was examined by immunocytochemistry. We investigated the functional consequences of inhibiting glutamate uptake by recording cochlear potentials during intracochlear perfusion with either l-trans-pyrrolidine-2,4-dicarboxylic acid or dihydrokainate. At the end of the electrophysiological session, cochleas were processed for electron microscopy. Only the glutamate aspartate transporter messenger ribonucleic acid was detected in the organ of Corti. Consistently, glutamate aspartate transporter protein was detected in the inner hair cell-supporting cells and in the ganglion of Corti satellite cells. Glutamate transporter and excitatory amino acid carrier were found in the afferent auditory neurons. Only intracochlear perfusions with l-trans-pyrrolidine-2,4-dicarboxylic acid resulted in a dose-dependent decrease in the amplitude of the cochlear compound action potential, leaving cochlear microphonic potential unaffected. After l-trans-pyrrolidine-2,4-dicarboxylic acid perfusion, cochleas displayed a swelling of the afferent endings typical of excitotoxicity. [(-)1-(4-aminophenyl)-4-methyl-7,8-methylenedioxy-4,5-dihydro-3-methylcarb amyl-2,3-benzodiazepine], a selective alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptor antagonist protects the cochlea against l-trans-pyrrolidine-2,4-dicarboxylic acid effect.
机译：在耳蜗中，谷氨酸在内部毛细胞和初级听觉神经元之间的突触传递中起主要作用。必须调节细胞外谷氨酸的浓度以防止兴奋性中毒。这种调节是由兴奋性氨基酸转运蛋白介导的，该蛋白是从突触间隙去除谷氨酸的膜蛋白。在这项研究中，我们调查了豚鼠耳蜗中三种兴奋性氨基酸转运蛋白亚型的分布和活性：谷氨酸天冬氨酸转运蛋白，谷氨酸转运蛋白和兴奋性氨基酸载体。使用豚鼠脑互补脱氧核糖核酸作为模板，通过与简并引物的聚合酶链反应，确定这些转运蛋白各自的部分信使核糖核酸序列。然后设计针对每种转运蛋白的特异性引物，并将其用于筛选Corti互补脱氧核糖核酸文库的解剖器官。通过免疫细胞化学检查每个转运蛋白的细胞分布。我们通过在耳蜗内灌注1-反-吡咯烷-2,4-二羧酸或二氢海藻酸酯来记录耳蜗电位来研究抑制谷氨酸摄取的功能后果。在电生理学阶段结束时，对耳蜗进行电子显微镜检查。在Corti的器官中仅检测到谷氨酸天冬氨酸转运蛋白信使核糖核酸。一致地，在内部毛细胞支持细胞和Corti卫星细胞的神经节中检测到谷氨酸天冬氨酸转运蛋白。在传入听觉神经元中发现了谷氨酸转运蛋白和兴奋性氨基酸载体。仅用1-反式-吡咯烷-2，4-二羧酸进行的耳蜗内灌注导致耳蜗复合动作电位幅度的剂量依赖性降低，而使耳蜗微音电位不受影响。在l-反式-吡咯烷-2,4-二羧酸灌注后，耳蜗显示出典型的兴奋性毒性传入末梢的肿胀。 [（-）1-（4-氨基苯基）-4-甲基-7,8-亚甲基二氧基-4,5-二氢-3-甲基碳戊基-2,3-苯并二氮杂]]，选择性α-氨基-3-羟基- 5-甲基异恶唑-4-丙酸受体拮抗剂可保护耳蜗免受1-反-吡咯烷-2,4-二羧酸的影响。

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