Small molecule injection into single-cell C. elegans embryos via carbon-reinforced nanopipettes.

The introduction of chemical inhibitors into living cells at specific times in development is a useful method for investigating the roles of specific proteins or cytoskeletal components in developmental processes. Some embryos, such as those of Caenorhabditis elegans, however, possess a tough eggshe...

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Autores principales:	Lucy D Brennan, Thibault Roland, Diane G Morton, Shanna M Fellman, SueYeon Chung, Mohammad Soltani, Joshua W Kevek, Paul M McEuen, Kenneth J Kemphues, Michelle D Wang
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/703027dda1924cc7a745df32a6ea72fb
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spelling	oai:doaj.org-article:703027dda1924cc7a745df32a6ea72fb2021-11-18T08:53:37ZSmall molecule injection into single-cell C. elegans embryos via carbon-reinforced nanopipettes.1932-620310.1371/journal.pone.0075712https://doaj.org/article/703027dda1924cc7a745df32a6ea72fb2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24086620/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203The introduction of chemical inhibitors into living cells at specific times in development is a useful method for investigating the roles of specific proteins or cytoskeletal components in developmental processes. Some embryos, such as those of Caenorhabditis elegans, however, possess a tough eggshell that makes introducing drugs and other molecules into embryonic cells challenging. We have developed a procedure using carbon-reinforced nanopipettes (CRNPs) to deliver molecules into C. elegans embryos with high temporal control. The use of CRNPs allows for cellular manipulation to occur just subsequent to meiosis II with minimal damage to the embryo. We have used our technique to replicate classical experiments using latrunculin A to inhibit microfilaments and assess its effects on early polarity establishment. Our injections of latrunculin A confirm the necessity of microfilaments in establishing anterior-posterior polarity at this early stage, even when microtubules remain intact. Further, we find that latrunculin A treatment does not prevent association of PAR-2 or PAR-6 with the cell cortex. Our experiments demonstrate the application of carbon-reinforced nanopipettes to the study of one temporally-confined developmental event. The use of CRNPs to introduce molecules into the embryo should be applicable to investigations at later developmental stages as well as other cells with tough outer coverings.Lucy D BrennanThibault RolandDiane G MortonShanna M FellmanSueYeon ChungMohammad SoltaniJoshua W KevekPaul M McEuenKenneth J KemphuesMichelle D WangPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 9, p e75712 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Lucy D Brennan Thibault Roland Diane G Morton Shanna M Fellman SueYeon Chung Mohammad Soltani Joshua W Kevek Paul M McEuen Kenneth J Kemphues Michelle D Wang Small molecule injection into single-cell C. elegans embryos via carbon-reinforced nanopipettes.
description	The introduction of chemical inhibitors into living cells at specific times in development is a useful method for investigating the roles of specific proteins or cytoskeletal components in developmental processes. Some embryos, such as those of Caenorhabditis elegans, however, possess a tough eggshell that makes introducing drugs and other molecules into embryonic cells challenging. We have developed a procedure using carbon-reinforced nanopipettes (CRNPs) to deliver molecules into C. elegans embryos with high temporal control. The use of CRNPs allows for cellular manipulation to occur just subsequent to meiosis II with minimal damage to the embryo. We have used our technique to replicate classical experiments using latrunculin A to inhibit microfilaments and assess its effects on early polarity establishment. Our injections of latrunculin A confirm the necessity of microfilaments in establishing anterior-posterior polarity at this early stage, even when microtubules remain intact. Further, we find that latrunculin A treatment does not prevent association of PAR-2 or PAR-6 with the cell cortex. Our experiments demonstrate the application of carbon-reinforced nanopipettes to the study of one temporally-confined developmental event. The use of CRNPs to introduce molecules into the embryo should be applicable to investigations at later developmental stages as well as other cells with tough outer coverings.
format	article
author	Lucy D Brennan Thibault Roland Diane G Morton Shanna M Fellman SueYeon Chung Mohammad Soltani Joshua W Kevek Paul M McEuen Kenneth J Kemphues Michelle D Wang
author_facet	Lucy D Brennan Thibault Roland Diane G Morton Shanna M Fellman SueYeon Chung Mohammad Soltani Joshua W Kevek Paul M McEuen Kenneth J Kemphues Michelle D Wang
author_sort	Lucy D Brennan
title	Small molecule injection into single-cell C. elegans embryos via carbon-reinforced nanopipettes.
title_short	Small molecule injection into single-cell C. elegans embryos via carbon-reinforced nanopipettes.
title_full	Small molecule injection into single-cell C. elegans embryos via carbon-reinforced nanopipettes.
title_fullStr	Small molecule injection into single-cell C. elegans embryos via carbon-reinforced nanopipettes.
title_full_unstemmed	Small molecule injection into single-cell C. elegans embryos via carbon-reinforced nanopipettes.
title_sort	small molecule injection into single-cell c. elegans embryos via carbon-reinforced nanopipettes.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/703027dda1924cc7a745df32a6ea72fb
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Small molecule injection into single-cell C. elegans embryos via carbon-reinforced nanopipettes.

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