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Fat Body Organ Culture System in Aedes Aegypti, a Vector of Zika Virus
The insect fat body plays a central role in insect metabolism and nutrient storage, mirroring functions of the liver and fat tissue in vertebrates. Insect fat body tissue is usually distributed throughout the insect body. However, it is often concentrated in the abdomen and attached to the abdominal...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MyJove Corporation
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5614350/ https://www.ncbi.nlm.nih.gov/pubmed/28872112 http://dx.doi.org/10.3791/55508 |
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author | Chung, Hae-Na Rodriguez, Stacy D. Carpenter, Victoria K. Vulcan, Julia Bailey, C. Donovan Nageswara-Rao, Madhugiri Li, Yiyi Attardo, Geoffrey M. Hansen, Immo A. |
author_facet | Chung, Hae-Na Rodriguez, Stacy D. Carpenter, Victoria K. Vulcan, Julia Bailey, C. Donovan Nageswara-Rao, Madhugiri Li, Yiyi Attardo, Geoffrey M. Hansen, Immo A. |
author_sort | Chung, Hae-Na |
collection | PubMed |
description | The insect fat body plays a central role in insect metabolism and nutrient storage, mirroring functions of the liver and fat tissue in vertebrates. Insect fat body tissue is usually distributed throughout the insect body. However, it is often concentrated in the abdomen and attached to the abdominal body wall. The mosquito fat body is the sole source of yolk proteins, which are critical for egg production. Therefore, the in vitro culture of mosquito fat body tissues represents an important system for the study of mosquito physiology, metabolism, and, ultimately, egg production. The fat body culture process begins with the preparation of solutions and reagents, including amino acid stock solutions, Aedes physiological saline salt stock solution (APS), calcium stock solution, and fat body culture medium. The process continues with fat body dissection, followed by an experimental treatment. After treatment, a variety of different analyses can be performed, including RNA sequencing (RNA-Seq), qPCR, Western blots, proteomics, and metabolomics. In our example experiment, we demonstrate the protocol through the excision and culture of fat bodies from the yellow fever mosquito, Aedes aegypti, a principal vector of arboviruses including dengue, chikungunya, and Zika. RNA from fat bodies cultured under a physiological condition known to upregulate yolk proteins versus the control were subject to RNA-Seq analysis to demonstrate the potential utility of this procedure for investigations of gene expression. |
format | Online Article Text |
id | pubmed-5614350 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | MyJove Corporation |
record_format | MEDLINE/PubMed |
spelling | pubmed-56143502017-10-10 Fat Body Organ Culture System in Aedes Aegypti, a Vector of Zika Virus Chung, Hae-Na Rodriguez, Stacy D. Carpenter, Victoria K. Vulcan, Julia Bailey, C. Donovan Nageswara-Rao, Madhugiri Li, Yiyi Attardo, Geoffrey M. Hansen, Immo A. J Vis Exp Infectious Diseases The insect fat body plays a central role in insect metabolism and nutrient storage, mirroring functions of the liver and fat tissue in vertebrates. Insect fat body tissue is usually distributed throughout the insect body. However, it is often concentrated in the abdomen and attached to the abdominal body wall. The mosquito fat body is the sole source of yolk proteins, which are critical for egg production. Therefore, the in vitro culture of mosquito fat body tissues represents an important system for the study of mosquito physiology, metabolism, and, ultimately, egg production. The fat body culture process begins with the preparation of solutions and reagents, including amino acid stock solutions, Aedes physiological saline salt stock solution (APS), calcium stock solution, and fat body culture medium. The process continues with fat body dissection, followed by an experimental treatment. After treatment, a variety of different analyses can be performed, including RNA sequencing (RNA-Seq), qPCR, Western blots, proteomics, and metabolomics. In our example experiment, we demonstrate the protocol through the excision and culture of fat bodies from the yellow fever mosquito, Aedes aegypti, a principal vector of arboviruses including dengue, chikungunya, and Zika. RNA from fat bodies cultured under a physiological condition known to upregulate yolk proteins versus the control were subject to RNA-Seq analysis to demonstrate the potential utility of this procedure for investigations of gene expression. MyJove Corporation 2017-08-19 /pmc/articles/PMC5614350/ /pubmed/28872112 http://dx.doi.org/10.3791/55508 Text en Copyright © 2017, Journal of Visualized Experiments http://creativecommons.org/licenses/by/3.0/us/ This is an open-access article distributed under the terms of the Creative Commons Attribution 3.0 License. To view a copy of this license, visithttp://creativecommons.org/licenses/by/3.0/us/ |
spellingShingle | Infectious Diseases Chung, Hae-Na Rodriguez, Stacy D. Carpenter, Victoria K. Vulcan, Julia Bailey, C. Donovan Nageswara-Rao, Madhugiri Li, Yiyi Attardo, Geoffrey M. Hansen, Immo A. Fat Body Organ Culture System in Aedes Aegypti, a Vector of Zika Virus |
title | Fat Body Organ Culture System in Aedes Aegypti, a Vector of Zika Virus |
title_full | Fat Body Organ Culture System in Aedes Aegypti, a Vector of Zika Virus |
title_fullStr | Fat Body Organ Culture System in Aedes Aegypti, a Vector of Zika Virus |
title_full_unstemmed | Fat Body Organ Culture System in Aedes Aegypti, a Vector of Zika Virus |
title_short | Fat Body Organ Culture System in Aedes Aegypti, a Vector of Zika Virus |
title_sort | fat body organ culture system in aedes aegypti, a vector of zika virus |
topic | Infectious Diseases |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5614350/ https://www.ncbi.nlm.nih.gov/pubmed/28872112 http://dx.doi.org/10.3791/55508 |
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