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钙成像技术在疼痛机制研究中的应用

Investigation of Pain Mechanisms by Calcium Imaging Approaches.

机构信息

The Solomon H. Snyder Department of Neuroscience, Center for Sensory Biology, School of Medicine, Johns Hopkins University, Baltimore, MD, 21205, USA.

Howard Hughes Medical Institute, School of Medicine, Johns Hopkins University, Baltimore, MD, 21205, USA.

出版信息

Neurosci Bull. 2018 Feb;34(1):194-199. doi: 10.1007/s12264-017-0139-9. Epub 2017 May 13.

Abstract

Due to the complex circuitry and plethora of cell types involved in somatosensation, it is becoming increasingly important to be able to observe cellular activity at the population level. In addition, since cells rely on an intricate variety of extracellular factors, it is important to strive to maintain the physiological environment. Many electrophysiological techniques require the implementation of artificially-produced physiological environments and it can be difficult to assess the activity of many cells simultaneously. Moreover, imaging Ca transients using Ca-sensitive dyes often requires in vitro preparations or in vivo injections, which can lead to variable expression levels. With the development of more sensitive genetically-encoded Ca indicators (GECIs) it is now possible to observe changes in Ca transients in large populations of cells at the same time. Recently, groups have used a GECI called GCaMP to address fundamental questions in somatosensation. Researchers can now induce GCaMP expression in the mouse genome using viral or gene knock-in approaches and observe the activity of populations of cells in the pain pathway such as dorsal root ganglia (DRG), spinal neurons, or glia. This approach can be used in vivo and thus maintains the organism's biological integrity. The implementation of GCaMP imaging has led to many advances in our understanding of somatosensation. Here, we review the current findings in pain research using GCaMP imaging as well as discussing potential methodological considerations.

摘要

由于躯体感觉涉及到复杂的电路和大量的细胞类型,因此能够在群体水平上观察细胞活动变得越来越重要。此外,由于细胞依赖于复杂多样的细胞外因素,因此努力维持生理环境也很重要。许多电生理技术需要实施人为产生的生理环境,并且同时评估许多细胞的活性可能会很困难。此外,使用 Ca 敏感染料对 Ca 瞬变进行成像通常需要体外制备或体内注射,这可能导致表达水平的变化。随着更敏感的基因编码 Ca 指示剂 (GECI) 的发展,现在可以同时观察到大量细胞中 Ca 瞬变的变化。最近,一些研究小组使用一种称为 GCaMP 的 GECI 来解决躯体感觉中的基本问题。研究人员现在可以使用病毒或基因敲入方法在小鼠基因组中诱导 GCaMP 的表达,并观察疼痛通路中细胞群体的活动,如背根神经节 (DRG)、脊髓神经元或神经胶质细胞。这种方法可以在体内使用,从而保持生物体的生物完整性。GCaMP 成像的实施推动了我们对躯体感觉的理解的许多进展。在这里,我们回顾了使用 GCaMP 成像进行疼痛研究的最新发现,并讨论了潜在的方法学考虑因素。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6954/5799123/dccee34f30d2/12264_2017_139_Fig1_HTML.jpg

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