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实时快速循环伏安法测量预缺血预处理对大鼠尾状核脑片缺氧去极化引起的多巴胺外排的影响。

Effect of pre-ischaemic conditioning on hypoxic depolarization of dopamine efflux in the rat caudate brain slice measured in real-time with fast cyclic voltammetry.

机构信息

Division of Biomedical Science, St George's, University of London, London SW17 0RE, United Kingdom.

出版信息

Neurochem Int. 2011 Oct;59(5):714-21. doi: 10.1016/j.neuint.2011.06.020. Epub 2011 Jul 5.

Abstract

Fast cyclic voltammetry can be used to measure dopamine release after oxygen and glucose deprivation (OGD) induced anoxic depolarization in vitro. Here we measure dopamine efflux with 1s time resolution, which is appropriate to measure OGD-evoked dopamine efflux accurately. In the present study, we examined whether OGD-evoked dopamine efflux could be used to show pre-ischaemic conditioning in the rat caudate brain slice. Caudate slices were exposed to 0, 2, or 10 min OGD pre-ischaemic conditioning, then 60 min later exposed to a second OGD event of 15 min duration. We measured the OGD-evoked dopamine efflux using fast cyclic voltammetry and in some experiments caudate dopamine and DOPAC tissue levels were measured using HPLC and 20 μm cryostat sections were Nissl stained to indicate neuronal loss. We found that 10 but not 2 min OGD pre-ischaemic conditioning resulted in a longer time to onset of OGD-evoked dopamine efflux on the main OGD event (475 ± 31 and 287 ± 30 s for 10 Vs 0 min pre-ischaemic conditioning respectively). Further, 10 min OGD pre-ischaemic conditioning resulted in less dopamine efflux on the second OGD event (4.23 ± 1.12 and 8.14 ± 0.82 μM for 10 Vs 0 min pre-ischaemic conditioning respectively), despite these slices having similar tissue dopamine content and DOPAC/DA ratio, and the rate of dopamine release was slower in the main OGD event (21 ± 5 and 74 ± 8 nM/s for 10 Vs 0 min pre-ischaemic conditioning respectively). These data suggest that 10 min OGD pre-ischaemic conditioning can evoke tolerance to a second OGD event and that voltammetric recording of OGD-evoked dopamine efflux is a useful model of pre-ischaemic conditioning in neuronal tissue.

摘要

快速循环伏安法可用于测量体外氧葡萄糖剥夺(OGD)诱导缺氧去极化后多巴胺的释放。在这里,我们以 1s 的时间分辨率测量多巴胺外溢,这适合准确测量 OGD 诱发的多巴胺外溢。在本研究中,我们研究了 OGD 诱发的多巴胺外溢是否可用于显示大鼠尾状核脑片的缺血预处理。尾状核切片暴露于 0、2 或 10min 的 OGD 缺血预处理,然后 60min 后暴露于 15min 的第二次 OGD 事件。我们使用快速循环伏安法测量 OGD 诱发的多巴胺外溢,在一些实验中使用 HPLC 测量尾状核多巴胺和 DOPAC 组织水平,并使用 20μm 冷冻切片进行尼氏染色以指示神经元丢失。我们发现,10min 而非 2min 的 OGD 缺血预处理导致在主要 OGD 事件中 OGD 诱发的多巴胺外溢的起始时间延长(分别为 475±31 和 287±30s 用于 10 与 0min 缺血预处理)。此外,10min 的 OGD 缺血预处理导致第二次 OGD 事件中的多巴胺外溢减少(分别为 4.23±1.12 和 8.14±0.82μM 用于 10 与 0min 缺血预处理),尽管这些切片具有相似的组织多巴胺含量和 DOPAC/DA 比值,并且主要 OGD 事件中的多巴胺释放速度较慢(分别为 21±5 和 74±8nM/s 用于 10 与 0min 缺血预处理)。这些数据表明,10min 的 OGD 缺血预处理可以引发对第二次 OGD 事件的耐受,并且 OGD 诱发的多巴胺外溢的伏安法记录是神经元组织缺血预处理的有用模型。

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