@article{1c9c8875ba264e0d91c4c1ea52bd0686,
title = "Time course analysis of sensory axon regeneration in vivo by directly tracing regenerating axons",
abstract = "Most current studies quantify axon regeneration by immunostaining regeneration-associated proteins, representing indirect measurement of axon lengths from both sensory neurons in the dorsal root ganglia and motor neurons in the spinal cord. Our recently developed method of in vivo electroporation of plasmid DNA encoding for enhanced green fluorescent protein into adult sensory neurons in the dorsal root ganglia provides a way to directly and specifically measure regenerating sensory axon lengths in whole-mount nerves. A mouse model of sciatic nerve compression was established by squeezing the sciatic nerve with tweezers. Plasmid DNA carrying enhanced green fluorescent protein was transfected by ipsilateral dorsal root ganglion electroporation 2 or 3 days before injury. Fluorescence distribution of dorsal root or sciatic nerve was observed by confocal microscopy. At 12 and 18 hours, and 1, 2, 3, 4, 5, and 6 days of injury, lengths of regenerated axons after sciatic nerve compression were measured using green fluorescence images. Apoptosis-related protein caspase-3 expression in dorsal root ganglia was determined by western blot assay. We found that in vivo electroporation did not affect caspase-3 expression in dorsal root ganglia. Dorsal root ganglia and sciatic nerves were successfully removed and subjected to a rapid tissue clearing technique. Neuronal soma in dorsal root ganglia expressing enhanced green fluorescent protein or fluorescent dye-labeled microRNAs were imaged after tissue clearing. The results facilitate direct time course analysis of peripheral nerve axon regeneration. This study was approved by the Institutional Animal Care and Use Committee of Guilin Medical University, China (approval No. GLMC201503010) on March 7, 2014.",
keywords = "axon regeneration, cell apoptosis, dorsal root ganglion, in vivo electroporation, microRNAs, peripheral nervous system, sciatic nerve, tissue clearing",
author = "Yan Gao and Hu, {Yi Wen} and Duan, {Run Shan} and Yang, {Shu Guang} and Zhou, {Feng Quan} and Wang, {Rui Ying}",
note = "Funding Information: This work was supported by the National Natural Science Foundation of China, No. 81460198, 31260233; the National Institute of Health of the United States of American, No. R01NS064288, R01NS085176, R01EY027347 (to FQZ); the Craig H. Neilson Foundation, the BrightFocus Foundation (to FQZ). Funding Information: Author contributions: Study design, data collection and assembly, data analysis and interpretation, and manuscript writing: YG and YWH; data collection and assembly, data analysis and interpretation: SGY and RSD; study conception and design, manuscript writing, final approval of manuscript, and fundraising: RYW and FQZ. All authors approved the final version of the paper. Conflicts of interest: The authors declare that there is no conflict of interests. Financial support: This work was supported by the National Natural Science Foundation of China, No. 81460198, 31260233; the National Institute of Health of the United States of American, No. R01NS064288, R01NS085176, R01EY027347 (to FQZ); the Craig H. Neilson Foundation, the BrightFocus Foundation (to FQZ). Funders had no involvement in the study design; data col-lection, analysis, and interpretation; paper writing; or decision to submit the paper for publication. Institutional review board statement: The study was approved by the Institutional Animal Care and Use Committee of Guilin Medical University, China (approval No. GLMC201503010) on March 7, 2014. Copyright license agreement: The Copyright License Agreement has been signed by all authors before publication. Data sharing statement: Datasets analyzed during the current study are available from the corresponding author on reasonable request. Plagiarism check: Checked twice by iThenticate. Peer review: Externally peer reviewed. Open access statement: This is an open access journal, and articles are distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as appropriate credit is given and the new creations are licensed under the identical terms. Open peer reviewer: Andr{\'e} Poot, University of Twente, Netherlands. Publisher Copyright: {\textcopyright} 2020 Wolters Kluwer Medknow Publications. All rights reserved.",
year = "2020",
month = jun,
day = "1",
doi = "10.4103/1673-5374.270315",
language = "English (US)",
volume = "15",
pages = "1160--1165",
journal = "Neural Regeneration Research",
issn = "1673-5374",
publisher = "Editorial Board of Neural Regeneration Research",
number = "6",
}