Imaging pericytes and capillary diameter in brain slices and isolated retinae
Imaging pericytes and capillary diameter in brain slices and isolated retinae
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DOI：
10.1038/nprot.2014.019
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发表时间：
2014-02-01
期刊：
影响因子：
14.8
通讯作者：
Attwell, David
中科院分区：
文献类型：
--
作者：
Mishra, Anusha;O'Farrell, Fergus M.;Attwell, David
脑循环在结构和功能上都是高度专业化的，它为大脑的活动区域提供了微调的氧气和营养供应。我们对脑小动脉血流调节的理解发展迅速。最近的研究为微血管研究开辟了新的途径;例如，已经证明毛细血管壁上发现的收缩性周细胞会诱导毛细血管直径变化以响应神经递质，这表明周细胞可能在神经血管耦合中发挥作用。这一概念与传统的脑血流调节模型不一致，后者认为只有小动脉控制脑血流。在毛细血管水平的神经血管耦合的机制的调查需要一系列的方法，这涉及到独特的技术挑战。在这里，我们提供了详细的协议，成功的生理和免疫组织化学研究周细胞和毛细血管的脑切片和离体视网膜，使研究人员探测毛细血管的作用，神经血管耦合。该方案可在6-8小时内完成;然而，免疫组化实验可能需要3-6天。
The cerebral circulation is highly specialized, both structurally and functionally, and it provides a fine-tuned supply of oxygen and nutrients to active regions of the brain. Our understanding of blood flow regulation by cerebral arterioles has evolved rapidly. Recent work has opened new avenues in microvascular research; for example, it has been demonstrated that contractile pericytes found on capillary walls induce capillary diameter changes in response to neurotransmitters, suggesting that pericytes could have a role in neurovascular coupling. This concept is at odds with traditional models of brain blood flow regulation, which assume that only arterioles control cerebral blood flow. The investigation of mechanisms underlying neurovascular coupling at the capillary level requires a range of approaches, which involve unique technical challenges. Here we provide detailed protocols for the successful physiological and immunohistochemical study of pericytes and capillaries in brain slices and isolated retinae, allowing investigators to probe the role of capillaries in neurovascular coupling. This protocol can be completed within 6-8 h; however, immunohistochemical experiments may take 3-6 d.