Department of Neurology, University of Toledo College of Medicine, 3000 Arlington Avenue, Toledo, OH 43614, USA.
Neurochem Res. 2012 Apr;37(4):732-9. doi: 10.1007/s11064-011-0666-0. Epub 2011 Dec 7.
There is evidence from immunohistochemistry, quantitative microchemistry, and pharmacology for several amino acids as neurotransmitters in the vestibular nuclear complex (VNC), including glutamate, γ-aminobutyrate (GABA), and glycine. However, evidence from measurements of release has been limited. The purpose of this study was to measure depolarization-stimulated calcium-dependent release of amino acids from the VNC in brain slices. Coronal slices containing predominantly the VNC were prepared from rats and perfused with artificial cerebrospinal fluid (ACSF) in an interface chamber. Fluid was collected from the chamber just downstream from the VNC using a microsiphon. Depolarization was induced by 50 mM potassium in either control calcium and magnesium concentrations or reduced calcium and elevated magnesium. Amino acid concentrations in effluent fluid were measured by high performance liquid chromatography. Glutamate release increased fivefold during depolarization in control calcium concentration and twofold in low calcium/high magnesium. These same ratios were 6 and 1.5 for GABA, 2 and 1.3 for glycine, and 2 and 1.5 for aspartate. Differences between release in control and low calcium/high magnesium ACSF were statistically significant for glutamate, GABA, and glycine. Glutamine release decreased during and after depolarization, and taurine release slowly increased. No evidence for calcium-dependent release was found for serine, glutamine, alanine, threonine, arginine, taurine, or tyrosine. Our results support glutamate and GABA as major neurotransmitters in the VNC. They also support glycine as a neurotransmitter and some function for taurine.
有证据表明,几种氨基酸在前庭神经核复合体(VNC)中作为神经递质,包括谷氨酸、γ-氨基丁酸(GABA)和甘氨酸。然而,关于释放的证据一直有限。本研究的目的是测量脑片 VNC 中氨基酸在去极化刺激下钙依赖性释放的情况。从大鼠中制备包含主要 VNC 的冠状切片,并在界面室中用人工脑脊液(ACSF)灌注。使用微吸管从 VNC 下游的腔室中收集流体。在对照钙镁浓度或低钙高镁条件下,用 50mM 钾诱导去极化。通过高效液相色谱法测量流出液中氨基酸的浓度。在对照钙浓度下,谷氨酸的释放在去极化时增加了五倍,在低钙高镁条件下增加了两倍。GABA 和甘氨酸的释放比分别为 6 和 1.5,2 和 1.3。谷氨酸、GABA 和甘氨酸在对照和低钙高镁 ACSF 中的释放差异具有统计学意义。谷氨酰胺在去极化期间和之后减少,牛磺酸缓慢增加。丝氨酸、谷氨酰胺、丙氨酸、苏氨酸、精氨酸、牛磺酸或酪氨酸没有发现钙依赖性释放的证据。我们的结果支持谷氨酸和 GABA 作为 VNC 中的主要神经递质。它们还支持甘氨酸作为神经递质和牛磺酸的某些功能。