1] Department of Neuroscience, Physiology and Pharmacology, University College London, London, UK. [2].
Department of Neuroscience, Physiology and Pharmacology, University College London, London, UK.
Nat Protoc. 2014 Feb;9(2):323-36. doi: 10.1038/nprot.2014.019. Epub 2014 Jan 16.
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.
大脑循环在结构和功能上都高度专业化，为大脑活跃区域提供精细调节的氧气和营养供应。我们对脑小动脉血流调节的理解迅速发展。最近的工作为微血管研究开辟了新途径；例如，已经证明，在毛细血管壁上发现的收缩型周细胞对神经递质做出反应，诱导毛细血管直径发生变化，这表明周细胞在神经血管耦联中可能发挥作用。这一概念与传统的大脑血流调节模型不一致，传统模型假设只有小动脉控制脑血流。在毛细血管水平上研究神经血管耦联的机制需要一系列方法，这涉及到独特的技术挑战。在这里，我们提供了在脑切片和分离的视网膜中成功进行周细胞和毛细血管的生理学和免疫组织化学研究的详细方案，使研究人员能够探究毛细血管在神经血管耦联中的作用。该方案可在 6-8 小时内完成；然而，免疫组织化学实验可能需要 3-6 天。
