학술논문

Fat Body Organ Culture System in Aedes Aegypti, a Vector of Zika Virus.
Document Type
article
Source
Journal of visualized experiments : JoVE. 2017(126)
Subject
Fat Body
Animals
Aedes
Egg Proteins
Organ Culture Techniques
Insect Vectors
Gene Expression
Zika Virus
Infectious Diseases
Issue 126
Fat body
organ culture
mosquitoes
Aedes aegypti
nutrition
vitellogenesis
Rare Diseases
Biotechnology
Emerging Infectious Diseases
Genetics
Prevention
Vector-Borne Diseases
Cognitive Sciences
Biochemistry and Cell Biology
Psychology
Language
Abstract
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.