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A multi-trap microfluidic chip enabling longitudinal studies of nerve regeneration in Caenorhabditis elegans
Several sophisticated microfluidic devices have recently been proposed for femtosecond laser axotomy in the nematode C. elegans for immobilization of the animals for surgery to overcome time-consuming and labor-intensive manual processes. However, nerve regeneration studies require long-term recover...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5575036/ https://www.ncbi.nlm.nih.gov/pubmed/28852096 http://dx.doi.org/10.1038/s41598-017-10302-4 |
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author | Gokce, Sertan Kutal Hegarty, Evan Marley Mondal, Sudip Zhao, Peisen Ghorashian, Navid Hilliard, Massimo A. Ben-Yakar, Adela |
author_facet | Gokce, Sertan Kutal Hegarty, Evan Marley Mondal, Sudip Zhao, Peisen Ghorashian, Navid Hilliard, Massimo A. Ben-Yakar, Adela |
author_sort | Gokce, Sertan Kutal |
collection | PubMed |
description | Several sophisticated microfluidic devices have recently been proposed for femtosecond laser axotomy in the nematode C. elegans for immobilization of the animals for surgery to overcome time-consuming and labor-intensive manual processes. However, nerve regeneration studies require long-term recovery of the animals and multiple imaging sessions to observe the regeneration capabilities of their axons post-injury. Here we present a simple, multi-trap device, consisting of a single PDMS (polydimethylsiloxane) layer, which can immobilize up to 20 animals at the favorable orientation for optical access needed for precise laser surgery and high-resolution imaging. The new device, named “worm hospital” allows us to perform the entire nerve regeneration studies, including on-chip axotomy, post-surgery housing for recovery, and post-recovery imaging all on one microfluidic chip. Utilizing the worm hospital and analysis of mutants, we observed that most but not all neurodevelopmental genes in the Wnt/Frizzled pathway are important for regeneration of the two touch receptor neurons ALM and PLM. Using our new chip, we observed that the cwn-2 and cfz-2 mutations significantly reduced the reconnection possibilities of both neurons without any significant reduction in the regrowth lengths of the severed axons. We observed a similar regeneration phenotype with cwn-1 mutation in ALM neurons only. |
format | Online Article Text |
id | pubmed-5575036 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-55750362017-09-01 A multi-trap microfluidic chip enabling longitudinal studies of nerve regeneration in Caenorhabditis elegans Gokce, Sertan Kutal Hegarty, Evan Marley Mondal, Sudip Zhao, Peisen Ghorashian, Navid Hilliard, Massimo A. Ben-Yakar, Adela Sci Rep Article Several sophisticated microfluidic devices have recently been proposed for femtosecond laser axotomy in the nematode C. elegans for immobilization of the animals for surgery to overcome time-consuming and labor-intensive manual processes. However, nerve regeneration studies require long-term recovery of the animals and multiple imaging sessions to observe the regeneration capabilities of their axons post-injury. Here we present a simple, multi-trap device, consisting of a single PDMS (polydimethylsiloxane) layer, which can immobilize up to 20 animals at the favorable orientation for optical access needed for precise laser surgery and high-resolution imaging. The new device, named “worm hospital” allows us to perform the entire nerve regeneration studies, including on-chip axotomy, post-surgery housing for recovery, and post-recovery imaging all on one microfluidic chip. Utilizing the worm hospital and analysis of mutants, we observed that most but not all neurodevelopmental genes in the Wnt/Frizzled pathway are important for regeneration of the two touch receptor neurons ALM and PLM. Using our new chip, we observed that the cwn-2 and cfz-2 mutations significantly reduced the reconnection possibilities of both neurons without any significant reduction in the regrowth lengths of the severed axons. We observed a similar regeneration phenotype with cwn-1 mutation in ALM neurons only. Nature Publishing Group UK 2017-08-29 /pmc/articles/PMC5575036/ /pubmed/28852096 http://dx.doi.org/10.1038/s41598-017-10302-4 Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Gokce, Sertan Kutal Hegarty, Evan Marley Mondal, Sudip Zhao, Peisen Ghorashian, Navid Hilliard, Massimo A. Ben-Yakar, Adela A multi-trap microfluidic chip enabling longitudinal studies of nerve regeneration in Caenorhabditis elegans |
title | A multi-trap microfluidic chip enabling longitudinal studies of nerve regeneration in Caenorhabditis elegans |
title_full | A multi-trap microfluidic chip enabling longitudinal studies of nerve regeneration in Caenorhabditis elegans |
title_fullStr | A multi-trap microfluidic chip enabling longitudinal studies of nerve regeneration in Caenorhabditis elegans |
title_full_unstemmed | A multi-trap microfluidic chip enabling longitudinal studies of nerve regeneration in Caenorhabditis elegans |
title_short | A multi-trap microfluidic chip enabling longitudinal studies of nerve regeneration in Caenorhabditis elegans |
title_sort | multi-trap microfluidic chip enabling longitudinal studies of nerve regeneration in caenorhabditis elegans |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5575036/ https://www.ncbi.nlm.nih.gov/pubmed/28852096 http://dx.doi.org/10.1038/s41598-017-10302-4 |
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