C. elegans major fats are stored in vesicles distinct from lysosome-related organelles.
Genetic conservation allows ancient features of fat storage endocrine pathways to be explored in C. elegans. Multiple studies have used Nile red or BODIPY-labeled fatty acids to identify regulators of fat mass. When mixed with their food, E. coli bacteria, Nile red, and BODIPY-labeled fatty acids stain multiple spherical cellular structures in the C. elegans major fat storage organ, the intestine. However, here we demonstrate that, in the conditions previously reported, the lysosome-related organelles stained by Nile red and BODIPY-labeled fatty acids are not the C. elegans major fat storage compartment. We show that the major fat stores are contained in a distinct cellular compartment that is not stained by Nile red. Using biochemical assays, we validate oil red O staining as a method to assess major fat stores in C. elegans, allowing for efficient and accurate genetic and functional genomic screens for genes that control fat accumulation at the organismal level.
O'Rourke
Eyleen J
EJ
Department of Molecular Biology, Massachusetts General Hospital, Boston, MA 02114, USA.
Soukas
Alexander A
AA
Carr
Christopher E
CE
Ruvkun
Gary
G
D000818:Animals; D001391:Azo Compounds; D001896:Boron Compounds; D017173:Caenorhabditis elegans / Q000378:metabolism* / Q000648:ultrastructure; D022162:Cytoplasmic Vesicles / Q000378:metabolism* / Q000648:ultrastructure; D005223:Fats / Q000378:metabolism*; D005456:Fluorescent Dyes; D010078:Oxazines; D013194:Staining and Labeling
C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001391:Azo Compounds; D001896:Boron Compounds; D005223:Fats; D005456:Fluorescent Dyes; D010078:Oxazines; C011049:oil red O; C044808:nile red