Keywords:Lipid metabolism, lipases, atherosclerosis, inflammation, lipid signaling, leukocytes, diabetes, placenta
Research interest:Intracellular lipid droplet formation, neutral lipid hydrolysis, and lipid catabolism are tightly regulated processes, which involve intracellular hydrolases, enzymes involved in lipid biosynthesis, and regulatory proteins. Excessive lipid storage in lipid droplets is central to the pathogenesis of prevalent metabolic diseases, such as obesity, diabetes, and atherosclerosis. The research interests of our group focus on lipid and energy metabolism in non-adipose tissues and cells. Over the past years, we concentrated our efforts on the consequences of lipase deficiency on the regulation of lipid and energy metabolism [1, 2]. We utilize transgenic and knockout mouse models with loss or overexpression of lipases to investigate the impact of the respective enzymes on systemic lipid and energy metabolism and atherogenesis. In addition, we are interested in cell and tissue autonomous functions of the respective enzymes. The role of lipases in lipid metabolism and inflammation in the placenta and the fetus are incompletely understood. We have previously reported on lipase activities in human and murine placenta with extracellular lipases (lipoprotein lipase, endothelial lipase) being involved . The role of intracellular lipases, however, has not been addressed. We hypothesize that loss of adipose triglyceride lipase, hormone-sensitive lipase, monoglyceride lipase, and lysosomal acid lipase affects lipid
Collaborations within the DP-iDP:
Know-how and infrastructure of the research group:Dagmar Kratky is a biochemist and molecular biologist at the Institute of Molecular Biology and Biochemistry. Since 2006 she has been leading her own group, which currently comprises 1 senior scientist, 1 postdoctoral fellow, and 4 PhD students. Two technicians with permanent positions are financed by the MUG. Equipment and methodologies are available for gene cloning, DNA sequencing, real-time PCR analysis, heterologous expression of genes as well as gene silencing techniques and adenoviral infections, and an FPLC system for lipoprotein analysis. The local infrastructure includes cell culture facilities, an isotope laboratory, fluorescent and deconvolution microscopes, GC and HPLC systems for lipid analysis, a proteomics platform, and an XF24 Extracellular Flux Analyzer (Seahorse) for simultaneous measurements of oxygen consumption and extracellular acidification rates. Our animal facility houses more than 25 different mouse models.
Scientific concepts and techniques that students will learn in this laboratory:In D. Kratky’s laboratory, DP-iDP students will get insights into whole body lipid and energy metabolism and the consequences of lipid and energy overload in disease. In addition, they will get background knowledge on biochemistry and cell biology of lipid droplet formation. DP-iDP students will be trained in radioactive and non-radioactive labeling of lipids and lipoproteins. Tracer experiments will include uptake studies of fatty acids and glucose. Students will learn RNA and DNA methodologies including isolation of RNA from tissues and cells, determine mRNA expression by RT and real-time PCR, isolate full-length cDNAs, clone cDNAs into protein expression vectors, transfect mammalian cells with recombinant constructs and silence specific genes with siRNA techniques. Moreover, selected collegiates will perform hydrolase activity assays, lipid uptake and efflux experiments, determine lipid parameters in plasma, cells and tissues, examine lipoprotein profiles, and analyze mRNA and protein expression. Students under D. Kratky’s supervision will be trained in animal handling and breeding, tissue and cell isolation, dietary studies, injections, blood sampling, and perfusions.