Methods for Investigating Cell Division Mechanisms in C. elegans

Ian D. Wolff; Nikita S. Divekar; Sarah M. Wignall

doi:10.1007/978-1-0716-1904-9_2

Methods for Investigating Cell Division Mechanisms in C. elegans

Ian D. Wolff, Nikita S. Divekar, Sarah M. Wignall^*

^*Corresponding author for this work

Molecular Biosciences

Research output: Chapter in Book/Report/Conference proceeding › Chapter

5 Scopus citations

Abstract

The nematode Caenorhabditis elegans is a widely used model organism for the study of mitotic and meiotic cell division. These self-fertilizing worms are particularly advantageous for such studies because they rapidly reproduce (each worm lays ~250 eggs in only 3–4 days) and the cell division machinery is highly conserved between worms and humans. Worms are also genetically tractable and proteins can be readily depleted using RNA interference (RNAi), allowing for the characterization of protein function in vivo. To assess phenotypes, spindles can be directly visualized within the worm using fluorescent protein tags or embryos can be dissected out of the worm and immunostained. A combination of these techniques allows comprehensive characterization of a protein’s function in a relatively short time span. Here, we describe methods for each of these techniques: RNA interference through feeding, in utero live imaging, in utero fixed imaging, and immunofluorescence.

Original language	English (US)
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	19-35
Number of pages	17
DOIs	https://doi.org/10.1007/978-1-0716-1904-9_2
State	Published - 2022

Publication series

Name	Methods in Molecular Biology
Volume	2415
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Funding

The authors thank Wignall Lab members past and present for their contributions in developing these protocols, especially Gabriel Cavin-Meza, Amanda Davis-Roca, Carissa Heath, Jeremy Hollis, Hannah Horton, Tim Mullen, Christina Muscat, and Michael Tran. This work was supported by American Heart Association Predoctoral Fellowship 17PRE33440016 (to I.D.W.), NIH/-NIGMS Molecular Biophysics Training Grant T32GM008382 (to I.D.W.), and NIH R01GM124354 (to S.M.W.). Microscopy was performed at the Biological Imaging Facility at Northwestern University, supported by the Chemistry for Life Processes Institute, the NU Office for Research, the Department of Molecular Bios-ciences, and the Rice Foundation. The authors thank Wignall Lab members past and present for their contributions in developing these protocols, especially Gabriel Cavin-Meza, Amanda Davis-Roca, Carissa Heath, Jeremy Hollis, Hannah Horton, Tim Mullen, Christina Muscat, and Michael Tran. This work was supported by American Heart Association Predoctoral Fellowship 17PRE33440016 (to I.D.W.), NIH/-NIGMS Molecular Biophysics Training Grant T32GM008382 (to I.D.W.), and NIH R01GM124354 (to S.M.W.). Microscopy was performed at the Biological Imaging Facility at Northwestern University, supported by the Chemistry for Life Processes Institute, the NU Office for Research, the Department of Molecular Biosciences, and the Rice Foundation.

Keywords

GFP
Meiosis
Microscopy
Oocyte
RNAi

ASJC Scopus subject areas

Genetics
Molecular Biology

Access to Document

10.1007/978-1-0716-1904-9_2

Cite this

@inbook{490b84b54f604c0d86674fc2a4947258,

title = "Methods for Investigating Cell Division Mechanisms in C. elegans",

abstract = "The nematode Caenorhabditis elegans is a widely used model organism for the study of mitotic and meiotic cell division. These self-fertilizing worms are particularly advantageous for such studies because they rapidly reproduce (each worm lays ~250 eggs in only 3–4 days) and the cell division machinery is highly conserved between worms and humans. Worms are also genetically tractable and proteins can be readily depleted using RNA interference (RNAi), allowing for the characterization of protein function in vivo. To assess phenotypes, spindles can be directly visualized within the worm using fluorescent protein tags or embryos can be dissected out of the worm and immunostained. A combination of these techniques allows comprehensive characterization of a protein{\textquoteright}s function in a relatively short time span. Here, we describe methods for each of these techniques: RNA interference through feeding, in utero live imaging, in utero fixed imaging, and immunofluorescence.",

keywords = "GFP, Meiosis, Microscopy, Oocyte, RNAi",

author = "Wolff, {Ian D.} and Divekar, {Nikita S.} and Wignall, {Sarah M.}",

note = "Publisher Copyright: {\textcopyright} 2022, Springer Science+Business Media, LLC, part of Springer Nature.",

year = "2022",

doi = "10.1007/978-1-0716-1904-9_2",

language = "English (US)",

series = "Methods in Molecular Biology",

publisher = "Humana Press Inc.",

pages = "19--35",

booktitle = "Methods in Molecular Biology",

}

TY - CHAP

T1 - Methods for Investigating Cell Division Mechanisms in C. elegans

AU - Wolff, Ian D.

AU - Divekar, Nikita S.

AU - Wignall, Sarah M.

PY - 2022

Y1 - 2022

N2 - The nematode Caenorhabditis elegans is a widely used model organism for the study of mitotic and meiotic cell division. These self-fertilizing worms are particularly advantageous for such studies because they rapidly reproduce (each worm lays ~250 eggs in only 3–4 days) and the cell division machinery is highly conserved between worms and humans. Worms are also genetically tractable and proteins can be readily depleted using RNA interference (RNAi), allowing for the characterization of protein function in vivo. To assess phenotypes, spindles can be directly visualized within the worm using fluorescent protein tags or embryos can be dissected out of the worm and immunostained. A combination of these techniques allows comprehensive characterization of a protein’s function in a relatively short time span. Here, we describe methods for each of these techniques: RNA interference through feeding, in utero live imaging, in utero fixed imaging, and immunofluorescence.

AB - The nematode Caenorhabditis elegans is a widely used model organism for the study of mitotic and meiotic cell division. These self-fertilizing worms are particularly advantageous for such studies because they rapidly reproduce (each worm lays ~250 eggs in only 3–4 days) and the cell division machinery is highly conserved between worms and humans. Worms are also genetically tractable and proteins can be readily depleted using RNA interference (RNAi), allowing for the characterization of protein function in vivo. To assess phenotypes, spindles can be directly visualized within the worm using fluorescent protein tags or embryos can be dissected out of the worm and immunostained. A combination of these techniques allows comprehensive characterization of a protein’s function in a relatively short time span. Here, we describe methods for each of these techniques: RNA interference through feeding, in utero live imaging, in utero fixed imaging, and immunofluorescence.

KW - GFP

KW - Meiosis

KW - Microscopy

KW - Oocyte

KW - RNAi

UR - http://www.scopus.com/inward/record.url?scp=85122456645&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85122456645&partnerID=8YFLogxK

U2 - 10.1007/978-1-0716-1904-9_2

DO - 10.1007/978-1-0716-1904-9_2

M3 - Chapter

C2 - 34972943

AN - SCOPUS:85122456645

T3 - Methods in Molecular Biology

SP - 19

EP - 35

BT - Methods in Molecular Biology

PB - Humana Press Inc.

ER -

Methods for Investigating Cell Division Mechanisms in C. elegans

Abstract

Publication series

Funding

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this