Biology Reference
In-Depth Information
CHAPTER
Drosophila melanogaster
4
Han Lee
*
, Ying Peng
{
, and Yi Guo
{,
{
Analysis of Lipid Droplet
Dynamics and Functions in
*
Graduate Program in Neurobiology of Disease, Mayo Clinic, Rochester, Minnesota, USA
{
Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota, USA
{
Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, Minnesota, USA
CHAPTER OUTLINE
4.1 Genome-Wide RNA Interference Screens in Drosophila Cells to Identify Regulators
for LD Biology .................................................................................................... 54
4.1.1 S2 Cells and their Advantage for RNAi Screens................................... 54
4.1.2 RNAi Library .................................................................................... 55
4.1.3 Screening Materials .......................................................................... 55
4.1.4 Screening Procedure......................................................................... 55
4.1.5 Screening Results and Discussion...................................................... 56
4.1.5.1 Class III Arf1/COPI Complex Regulates LD Breakdown.................58
4.1.5.2 Class V Phosphatidylcholine Biosynthesis Genes Regulate
LD Size ...................................................................................................58
4.2 LD Dynamics in Different Drosophila Developmental Stages .................................. 59
4.2.1 Genetic Manipulations of Fat Bodies for LD Study ............................... 61
4.2.2 Imaging LDs in Fixed Fat Bodies Using Confocal Microscopy................ 63
4.2.3 Imaging LD Dynamics in Vivo Using Coherent Anti-Stokes Raman
Scattering Microscopy ....................................................................63
4.3 LD Functions in Drosophila Physiology ................................................................ 64
4.3.1 LD Structural Proteins ...................................................................... 64
4.3.2 Lipases............................................................................................ 65
4.3.3 d Seipin ........................................................................................... 65
Acknowledgments ..................................................................................................... 66
References ............................................................................................................... 66
Abstract
The lipid droplet (LD) is a unique cellular organelle containing a neutral-lipid core
enclosed by a phospholipid monolayer and associated proteins. Despite the important
