Computational Analysis of the Caenorhabditis elegans Germline to Study the Distribution of Nuclei, Proteins, and the Cytoskeleton
Gopal, Sandeep LU
and Pocock, Roger
(2018)
In Journal of visualized experiments : JoVE
- Abstract
The Caenorhabditis elegans (C. elegans) germline is used to study several biologically important processes including stem cell development, apoptosis, and chromosome dynamics. While the germline is an excellent model, the analysis is often two dimensional due to the time and labor required for three-dimensional analysis. Major readouts in such studies are the number/position of nuclei and protein distribution within the germline. Here, we present a method to perform automated analysis of the germline using confocal microscopy and computational approaches to determine the number and position of nuclei in each region of the germline. Our method also analyzes germline protein distribution that enables the three-dimensional examination of... (More)
The Caenorhabditis elegans (C. elegans) germline is used to study several biologically important processes including stem cell development, apoptosis, and chromosome dynamics. While the germline is an excellent model, the analysis is often two dimensional due to the time and labor required for three-dimensional analysis. Major readouts in such studies are the number/position of nuclei and protein distribution within the germline. Here, we present a method to perform automated analysis of the germline using confocal microscopy and computational approaches to determine the number and position of nuclei in each region of the germline. Our method also analyzes germline protein distribution that enables the three-dimensional examination of protein expression in different genetic backgrounds. Further, our study shows variations in cytoskeletal architecture in distinct regions of the germline that may accommodate specific spatial developmental requirements. Finally, our method enables automated counting of the sperm in the spermatheca of each germline. Taken together, our method enables rapid and reproducible phenotypic analysis of the C. elegans germline.
(Less)
- author
- Gopal, Sandeep
LU
and Pocock, Roger
- publishing date
- 2018
- type
- Contribution to journal
- publication status
- published
- keywords
- Animals, Caenorhabditis elegans/genetics, Caenorhabditis elegans Proteins/genetics, Cell Nucleus/metabolism, Cytoskeleton/metabolism, Germ Cells, Male, Microtubules/metabolism
- in
- Journal of visualized experiments : JoVE
- issue
- 134
- article number
- e57702
- publisher
- JoVE
- external identifiers
-
- scopus:85046629417
- pmid:29733323
- ISSN
- 1940-087X
- DOI
- 10.3791/57702
- language
- English
- LU publication?
- no
- id
- dfb59a0b-5d0b-43bb-bd3b-dfbe8d942c94
- date added to LUP
- 2021-10-25 13:07:46
- date last changed
- 2024-04-20 14:56:07
@article{dfb59a0b-5d0b-43bb-bd3b-dfbe8d942c94,
abstract = {{<p>The Caenorhabditis elegans (C. elegans) germline is used to study several biologically important processes including stem cell development, apoptosis, and chromosome dynamics. While the germline is an excellent model, the analysis is often two dimensional due to the time and labor required for three-dimensional analysis. Major readouts in such studies are the number/position of nuclei and protein distribution within the germline. Here, we present a method to perform automated analysis of the germline using confocal microscopy and computational approaches to determine the number and position of nuclei in each region of the germline. Our method also analyzes germline protein distribution that enables the three-dimensional examination of protein expression in different genetic backgrounds. Further, our study shows variations in cytoskeletal architecture in distinct regions of the germline that may accommodate specific spatial developmental requirements. Finally, our method enables automated counting of the sperm in the spermatheca of each germline. Taken together, our method enables rapid and reproducible phenotypic analysis of the C. elegans germline.</p>}},
author = {{Gopal, Sandeep and Pocock, Roger}},
issn = {{1940-087X}},
keywords = {{Animals; Caenorhabditis elegans/genetics; Caenorhabditis elegans Proteins/genetics; Cell Nucleus/metabolism; Cytoskeleton/metabolism; Germ Cells; Male; Microtubules/metabolism}},
language = {{eng}},
number = {{134}},
publisher = {{JoVE}},
series = {{Journal of visualized experiments : JoVE}},
title = {{Computational Analysis of the Caenorhabditis elegans Germline to Study the Distribution of Nuclei, Proteins, and the Cytoskeleton}},
url = {{http://dx.doi.org/10.3791/57702}},
doi = {{10.3791/57702}},
year = {{2018}},
}