Laser-based single-axon transection for high-content axon injury and regeneration studies.

The investigation of the regenerative response of the neurons to axonal injury is essential to the development of new axoprotective therapies. Here we study the retinal neuronal RGC-5 cell line after laser transection, demonstrating that the ability of these cells to initiate a regenerative response...

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Autores principales:	Darío Kunik, Carolyne Dion, Tsuneyuki Ozaki, Leonard A Levin, Santiago Costantino
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/ab2f5fb9e5a24d3daed97b16352b228b
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spelling	oai:doaj.org-article:ab2f5fb9e5a24d3daed97b16352b228b2021-11-18T07:35:10ZLaser-based single-axon transection for high-content axon injury and regeneration studies.1932-620310.1371/journal.pone.0026832https://doaj.org/article/ab2f5fb9e5a24d3daed97b16352b228b2011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22073205/?tool=EBIhttps://doaj.org/toc/1932-6203The investigation of the regenerative response of the neurons to axonal injury is essential to the development of new axoprotective therapies. Here we study the retinal neuronal RGC-5 cell line after laser transection, demonstrating that the ability of these cells to initiate a regenerative response correlates with axon length and cell motility after injury. We show that low energy picosecond laser pulses can achieve transection of unlabeled single axons in vitro and precisely induce damage with micron precision. We established the conditions to achieve axon transection, and characterized RGC-5 axon regeneration and cell body response using time-lapse microscopy. We developed an algorithm to analyze cell trajectories and established correlations between cell motility after injury, axon length, and the initiation of the regeneration response. The characterization of the motile response of axotomized RGC-5 cells showed that cells that were capable of repair or regrowth of damaged axons migrated more slowly than cells that could not. Moreover, we established that RGC-5 cells with long axons could not recover their injured axons, and such cells were much more motile. The platform we describe allows highly controlled axonal damage with subcellular resolution and the performance of high-content screening in cell cultures.Darío KunikCarolyne DionTsuneyuki OzakiLeonard A LevinSantiago CostantinoPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 11, p e26832 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Darío Kunik Carolyne Dion Tsuneyuki Ozaki Leonard A Levin Santiago Costantino Laser-based single-axon transection for high-content axon injury and regeneration studies.
description	The investigation of the regenerative response of the neurons to axonal injury is essential to the development of new axoprotective therapies. Here we study the retinal neuronal RGC-5 cell line after laser transection, demonstrating that the ability of these cells to initiate a regenerative response correlates with axon length and cell motility after injury. We show that low energy picosecond laser pulses can achieve transection of unlabeled single axons in vitro and precisely induce damage with micron precision. We established the conditions to achieve axon transection, and characterized RGC-5 axon regeneration and cell body response using time-lapse microscopy. We developed an algorithm to analyze cell trajectories and established correlations between cell motility after injury, axon length, and the initiation of the regeneration response. The characterization of the motile response of axotomized RGC-5 cells showed that cells that were capable of repair or regrowth of damaged axons migrated more slowly than cells that could not. Moreover, we established that RGC-5 cells with long axons could not recover their injured axons, and such cells were much more motile. The platform we describe allows highly controlled axonal damage with subcellular resolution and the performance of high-content screening in cell cultures.
format	article
author	Darío Kunik Carolyne Dion Tsuneyuki Ozaki Leonard A Levin Santiago Costantino
author_facet	Darío Kunik Carolyne Dion Tsuneyuki Ozaki Leonard A Levin Santiago Costantino
author_sort	Darío Kunik
title	Laser-based single-axon transection for high-content axon injury and regeneration studies.
title_short	Laser-based single-axon transection for high-content axon injury and regeneration studies.
title_full	Laser-based single-axon transection for high-content axon injury and regeneration studies.
title_fullStr	Laser-based single-axon transection for high-content axon injury and regeneration studies.
title_full_unstemmed	Laser-based single-axon transection for high-content axon injury and regeneration studies.
title_sort	laser-based single-axon transection for high-content axon injury and regeneration studies.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/ab2f5fb9e5a24d3daed97b16352b228b
work_keys_str_mv	AT dariokunik laserbasedsingleaxontransectionforhighcontentaxoninjuryandregenerationstudies AT carolynedion laserbasedsingleaxontransectionforhighcontentaxoninjuryandregenerationstudies AT tsuneyukiozaki laserbasedsingleaxontransectionforhighcontentaxoninjuryandregenerationstudies AT leonardalevin laserbasedsingleaxontransectionforhighcontentaxoninjuryandregenerationstudies AT santiagocostantino laserbasedsingleaxontransectionforhighcontentaxoninjuryandregenerationstudies
_version_	1718423241655058432

Laser-based single-axon transection for high-content axon injury and regeneration studies.

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