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Maria Kokkinopoulou and Jrgen Markl (Johannes Gutenberg Universitt Mainz, Johannes-von-Mller-Weg 6, 55099, Mainz, Germany)
Rhogocytes are pore cells scattered throughout the connective tissues of diverse body parts of gastropods. Our preliminary studies from plastic embedded samples provided an overview of rhogocyte structure after 3D reconstruction. Their plasma membrane reveals tubular and vesicular invaginations, bridged by cytoplasmic bars that form distinctive slits of 20nm width. They are surrounded by an external basal lamina-like structure and the rough endoplasmic reticulum is the most prominent cellular structure. In-situ hybridization and immuno-gold labeling experiments in the planorbid snail Biomphalaria glabrata will affirm whether rhogocytes are the site of hemoglobin biosynthesis, the multimeric respiratory protein of this species. Next, we use cryo-electron tomography to collect data in close-to-native conditions. Samples from the pond snail Lymnaea stagnalis that expresses hemocyanin, and from the planorbid snail Biomphalaria glabrata will be used. This approach will reveal information about the existence of different types of rhogocytes and set light into the mechanism of hemocyanin and hemoglobin secretion into the hemolymph and the function of these cells in other pathways.