Effects of glucose and glutamine concentration in the formulation of the artificial cerebrospinal fluid (ACSF)
Effects of glucose and glutamine concentration in the formulation of the artificial cerebrospinal fluid (ACSF)
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DOI：
10.1016/j.brainres.2008.04.007
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发表时间：
2008-07-07
期刊：
影响因子：
2.9
通讯作者：
Bikson, Marom
中科院分区：
文献类型：
--
作者：
An, Je Hi;Su, Yuzhuo;Bikson, Marom
ACSF的组成是控制脑切片制备的细胞外环境的基础。“典型的”制剂缺乏氨基酸，并且含有比脑脊髓液（0.47 - 4.4 mM）更高浓度的葡萄糖（10 mM）。我们研究了不同浓度的谷氨酰胺（CSF中最丰富的氨基酸）和葡萄糖对大鼠海马脑片生理学的影响。将双极配对脉冲刺激应用于Schaffer侧支，并在CA 1锥体层中监测群体尖峰，持续约1小时。添加谷氨酰胺（0.5 m4）与10 mM的葡萄糖灌流的切片没有提高人口尖峰幅度。较高浓度的谷氨酰胺（2-5 mM）导致扩散抑制。在不存在谷氨酰胺的情况下，将葡萄糖浓度从10 mM降低至5 mM，减弱了群体峰值。在没有谷氨酰胺的情况下，将葡萄糖降低至2 mM，抑制了诱发的群体尖峰。用含有“生理”浓度葡萄糖（2 m4）和谷氨酰胺（0.S mm）的ACSF超灌注脑切片同样抑制了群体峰值。在单独的实验中，在高钾诱导的癫痫样活动，谷氨酰胺（0.5 mM）不影响爆发的持续时间，频率或波形。这些结果表明，ACSF中的葡萄糖浓度应保守地为10 mM，以使成对脉冲群体反应最大化，而生理浓度的谷氨酰胺（0.5 mM）的存在对成对脉冲反应和高K '诱导的癫痫素活性的影响最小。这些结果在体外脑切片灌流和体内脑灌流之间的根本差异的背景下进行了讨论。(c)2008 Elsevier B. V.保留所有权利。
The composition of the ACSF is fundamental in controlling the extracellular environment of the brain slice preparation. 'Typical' formulations lack amino acids and contain a higher concentration of glucose (10 mM) than in the cerebrospinal fluid (0.47 - 4.4 mM). We examined the effects of different concentrations of glutamine, the most abundant amino acid in the CSF, and glucose on rat hippocampal slice physiology. Bipolar paired-pulse stimulation was applied to the Schaffer collaterals and population spikes were monitored in the CA1 pyramidal layer for similar to 1 hour. Addition of glutamine (0.5 m4 to slices superfused with 10 mM of glucose did not enhance population spike amplitude. Higher concentration of glutamine (2-5 mM) resulted in spreading- depression. Decreasing glucose concentration from 10 mM to 5 mM, in the absence of glutamine, attenuated population spikes. Decreasing glucose to 2 mM, in the absence of glutamine, suppressed evoked population spikes. Superfusing brain slices with ACSF containing 'physiological' concentrations of both glucose (2 m4 and glutamine (O.S mm) similarly suppressed population spikes. In separate experiments, during high-K' induced epileptiform activity, glutamine (0.5 mM) did not affect the burst duration, frequency or waveform. These results suggest that the concentration of glucose in ACSF should conservatively be 10 mM in order to maximize paired-pulse population responses while the presence of physiological concentration of glutamine (0.5 mM) has minimal effects on pairedpulse responses and high-K' induced epileptiforin activity. These results are discussed in the context of fundamental differences between in vitro brain slice superfusion and in vivo brain perfusion. (c) 2008 Elsevier B.V. All rights reserved.