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多模原子力显微镜研究星形胶质细胞的电学性质

A case study of the electrical properties of astrocytes by multimode AFM.

机构信息

Chinese Academy of Sciences, Ningbo Institute of Materials Technology & Engineering, Ningbo, Zhejiang, China.

College of Food and Pharmaceutical Sciences, Ningbo University, Ningbo, China.

出版信息

J Microsc. 2019 Aug;275(2):75-81. doi: 10.1111/jmi.12804. Epub 2019 May 17.

DOI:10.1111/jmi.12804
PMID:31074501
Abstract

Astrocytes play an important role in the physiological functions of the central nervous system. In this study, contact potential differences (CPD) and capacitance gradients of the cell bodies and glial filaments of astrocytes were measured. Charge propagation properties in the astrocyte gap junctions were also studied using multimode Atomic Force Microscopy (AFM) at nanometre resolution. The CPD of the cell bodies and glial filaments were 324.2 ± 138.4 and 119.1 ± 31.7 mV, respectively. The measured capacitance gradients were 1.51 ± 0.31 and 1.98 ± 0.32 zF nm , respectively. The gap junctions in the astrocytes showed no charge propagation and were not electrically sensitive. This furthers our understanding of astrocytes and other types of neuroglia. LAY DESCRIPTION: Neuroglia cells play important structural and functional roles in central nervous system (CNS). Neuroglia cells exceed the number of neurons by 10∼50 and can be divided into macroglia and microglia. Astrocytes are macroglia and are the largest and most abundant cells in the CNS. Astrocytes lack axons and dendrites and do not propagate action potentials. They have few cytoplasmic organelles, but possess abundant glial filaments, the main components of the cytoskeleton. Glial filaments are composed of the glial fibrillary acidic protein (GFAP). Astrocytes produce intercellular calcium waves in their gap junctions mediated through receptor activator (such as glutamate) to permit signal transduction._ENREF_5 In addition to their role in the support and nutrition of neurons, astrocytes are involved in various types of CNS activity including: (1) cytokine secretion for neuronal survival, growth and differentiation; (2) protection from brain injury; (3) modulation of the blood brain barrier; and (4) neuronal immunity. Bidirectional crosstalk between the astrocytes and neurons exists. Astrocytes can be activated by neurotransmitters released and can themselves release gliotransmitters to act upon neurons. Astrocytes are closely related to various disease states, including epilepsy and Alzheimer's disease. In this study, the electrical properties of astrocytes, including the contact potential difference (CPD) and capacitance gradients of the cell bodies and glial filaments, and charge propagation in the gap junctions were investigated at the nanometer level using quantitative Kelvin Probe Force Microscopy (KPFM) and Electrostatic Force Microscopy (EFM). The CPD of the cell bodies and glial filaments of the astrocytes were 324.2 mV and 119.1 mV, respectively. Capacitance gradients of the cell bodies and glial filaments of the astrocytes were 1.51 zF/nm and 1.98 zF/nm, respectively. Gap junctions in the astrocytes do not perform charge propagation functions and the astrocytes are not electrically sensitive. One should note that these results from KPFM and EFM were measured on dried cell and the situation might be different when studying in operando environment, still these findings aid our understanding of the electrical properties and functions of astrocytes, and further our knowledge of the electrical properties of the CNS.

摘要

星形胶质细胞在中枢神经系统的生理功能中发挥着重要作用。在这项研究中,测量了星形胶质细胞胞体和神经胶质丝的接触电势差(CPD)和电容梯度。还使用多模原子力显微镜(AFM)在纳米分辨率下研究了星形胶质细胞缝隙连接中的电荷传播特性。胞体和神经胶质丝的 CPD 分别为 324.2±138.4mV 和 119.1±31.7mV。测量的电容梯度分别为 1.51±0.31zF·nm 和 1.98±0.32zF·nm。星形胶质细胞中的缝隙连接没有电荷传播,也没有电敏感性。这进一步加深了我们对星形胶质细胞和其他类型神经胶质的理解。
神经胶质细胞在中枢神经系统(CNS)中起着重要的结构和功能作用。神经胶质细胞的数量比神经元多 10∼50 倍,可分为大胶质细胞和小胶质细胞。星形胶质细胞是大胶质细胞,是中枢神经系统中最大和最丰富的细胞。星形胶质细胞缺乏轴突和树突,不能传播动作电位。它们的细胞质细胞器很少,但拥有丰富的神经胶质丝,这是细胞骨架的主要成分。神经胶质丝由神经胶质酸性蛋白(GFAP)组成。星形胶质细胞通过受体激活剂(如谷氨酸)在其缝隙连接中产生细胞间钙波,从而允许信号转导。
除了在神经元的支持和营养作用外,星形胶质细胞还参与各种类型的中枢神经系统活动,包括:(1)细胞因子分泌以促进神经元存活、生长和分化;(2)脑损伤保护;(3)调节血脑屏障;(4)神经元免疫。星形胶质细胞和神经元之间存在双向串扰。星形胶质细胞可以被释放的神经递质激活,也可以自身释放神经胶质递质作用于神经元。星形胶质细胞与各种疾病状态密切相关,包括癫痫和阿尔茨海默病。在这项研究中,使用定量 Kelvin 探针力显微镜(KPFM)和静电场力显微镜(EFM)在纳米尺度上研究了星形胶质细胞的电特性,包括胞体和神经胶质丝的接触电势差(CPD)和电容梯度,以及缝隙连接中的电荷传播。星形胶质细胞胞体和神经胶质丝的 CPD 分别为 324.2mV 和 119.1mV。星形胶质细胞胞体和神经胶质丝的电容梯度分别为 1.51zF/nm 和 1.98zF/nm。星形胶质细胞的缝隙连接没有电荷传播功能,星形胶质细胞没有电敏感性。值得注意的是,这些来自 KPFM 和 EFM 的结果是在干燥的细胞上测量的,在研究operando 环境时情况可能会有所不同,但这些发现有助于我们理解星形胶质细胞的电特性和功能,进一步了解中枢神经系统的电特性。

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