(518.3) From Flat to Bulb - Novel Insights in Caveolae Membrane Curvature

Poster Board Number: A429

Claudia Matthaeus (National Heart, Lung, and Blood Institute), Kem Sochacki (National Heart, Lung, and Blood Institute), Dmytro Puchkov (Leibniz Institute for Molecular Pharmacology), Volker Haucke (Leibniz Institute for Molecular Pharmacology), Martin Lehmann (Leibniz Institute for Molecular Pharmacology), Justin Taraska (National Heart, Lung, and Blood Institute)

Caveolae are 60-100 nm sized bulb-shaped plasma membrane invaginations found in various cell types including adipocytes, endothelial cells and myocytes. Their cellular functions comprise endocytosis and signaling processes, cellular lipid uptake, as well as sensing and regulation of membrane tension. Thereby, it was shown previously that caveolae can change their membrane shape from invaginated bulbs to flat domains and vice versa. However it is currently not clear which proteins are responsible for the membrane curvature changes. Here, we report the first detailed structural investigation of caveolae membrane shapes in vivo. To do so, Platinum replica EM images of the plasma membrane in mouse embryonic fibroblasts and endothelial cells were correlated to super-resolution (STED) fluorescence intensities of proteins associated with caveolae domains. This correlative imaging approach (CLEM) allowed us to precisely localize caveolae related proteins to the underlying membrane curvature. The major structural caveolae proteins such as Caveolins and Cavins were found in all caveolae membrane domains independent of their curvature. Surprisingly, EHD2, a caveolar neck protein, was not only detected at bulb-shaped invaginations but also at flat caveolae. Contrarily, Pacsin/Syndapin2 and EHBP1 were predominantly surrounding flat caveolae domains indicating that both proteins promote the neck formation. Because dynamin was previously associated with caveolae endocytosis and mobility we analyzed its spatial distribution to caveolae. However, our detailed CLEM analysis failed to determine dynamin to caveolae membrane domains in several cell types. Additionally, dynamin lacking cells did not show severe morphological changes of caveolae suggesting that dynamin is not involved in caveolae membrane curvature changes. Taken together, this study gives novel insights in caveolae structure and their formation from flat to invaginated membrane domains.