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利用病毒传递的 GCaMP6f 对小鼠周围神经中小直径纤维的电活动进行成像。

Imaging of electrical activity in small diameter fibers of the murine peripheral nerve with virally-delivered GCaMP6f.

机构信息

Department of Bioengineering, University of Colorado - Anschutz Medical Campus, Colorado, USA.

Department of Cell and Developmental Biology, University of Colorado - Anschutz Medical Campus, Colorado, USA.

出版信息

Sci Rep. 2018 Feb 19;8(1):3219. doi: 10.1038/s41598-018-21528-1.

Abstract

Current neural interfaces are hampered by lack of specificity and selectivity for neural interrogation. A method that might improve these interfaces is an optical peripheral nerve interface which communicates with individual axons via optogenetic reporters. To determine the feasibility of such an interface, we delivered the genetically encoded calcium indicator GCaMP6f to the mouse peripheral nerve by intramuscular injection of adenoassociated viral vector (AAV1) under the control of the CAG (chicken beta actin- cytomegalovirus hybrid promoter). Small diameter axons in the common peroneal nerve were transduced and demonstrated electrically inducible calcium transients ex vivo. Responses to single electrical stimuli were resolvable, and increasing the number of stimuli resulted in a monotonic increase in maximum fluorescence and a prolongation of calcium transient kinetics. This work demonstrates the viability of using a virally-delivered, genetically-encoded calcium indicator to read-out from peripheral nerve axons.

摘要

目前的神经接口受到缺乏神经询问的特异性和选择性的限制。一种可能改善这些接口的方法是光外周神经接口,它通过光遗传学报告器与单个轴突进行通信。为了确定这种接口的可行性,我们通过肌肉内注射腺相关病毒载体 (AAV1),在 CAG(鸡β肌动蛋白-巨细胞病毒杂交启动子)的控制下,将基因编码的钙指示剂 GCaMP6f 递送至小鼠外周神经。小直径轴突在坐骨神经中被转导,并在体外证明可诱导电钙瞬变。对单个电刺激的反应是可分辨的,并且增加刺激的数量导致最大荧光的单调增加和钙瞬变动力学的延长。这项工作证明了使用病毒传递的、基因编码的钙指示剂来读取外周神经轴突的可行性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b4ea/5818512/43ca5b630ff6/41598_2018_21528_Fig1_HTML.jpg

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