Dietary restriction during development enlarges intestinal and hypodermal lipid droplets in Caenorhabditis elegans.

Dietary restriction (DR) extends lifespan in man species and modulates evolutionary conserved signalling and metabolic pathways. Most of these studies were done in adult animals. Here we investigated fat phenotypes of C. elegans larvae and adults which were exposed to DR during development. This approach was named "developmental-DR" (dDR). Moderate as well as stringent dDR increased the triglyceride to protein ratio in L4 larvae and adult worms. This alteration was accompanied by a marked expansion of intestinal and hypodermal lipid droplets. In comparison to ad libitum condition, the relative proportion of fat stored in large lipid droplets (>50 µm(3)) was increased by a factor of about 5 to 6 in larvae exposed to dDR. Microarray-based expression profiling identified several dDR-regulated genes of lipolysis and lipogenesis which may contribute to the observed fat phenotypes. In conclusion, dDR increases the triglyceride to protein ratio, enlarges lipid droplets and alters the expression of genes functioning in lipid metabolism in C. elegans. These changes might be an effective adaptation to conserve fat stores in animals subjected to limiting food supply during development.

Fluorescence-based fixative and vital staining of lipid droplets in Caenorhabditis elegans reveal fat stores using microscopy and flow cytometry approaches.

The proportions of body fat and fat-free mass are determining factors of adiposity-associated diseases. Work in Caenorhabditis elegans has revealed evolutionarily conserved pathways of fat metabolism. Nevertheless, analysis of body composition and fat distribution in the nematodes has only been partially unraveled because of methodological difficulties. We characterized metabolic C. elegans mutants by using novel and feasible BODIPY 493/503-based fat staining and flow cytometry approaches. Fixative as well as vital BODIPY staining procedures visualize major fat stores, preserve native lipid droplet morphology, and allow quantification of fat content per body volume of individual worms. Colocalization studies using coherent anti-Stokes Raman scattering microscopy, Raman microspectroscopy, and imaging of lysosome-related organelles as well as biochemical measurement confirm our approaches. We found that the fat-to-volume ratio of dietary restriction, TGF-ß, and germline mutants are specific for each strain. In contrast, the proportion of fat-free mass is constant between the mutants, although their volumes differ by a factor of 3. Our approaches enable sensitive, accurate, and high-throughput assessment of adiposity in large C. elegans populations at a single-worm level.