Isolated bacteriocyte cell suspensions from the hydrothermal-vent tubeworm Riftia pachyptila, a potent tool for cellular physiology in a chemoautotrophic symbiosis

The hydrothermal-vent tubeworm Riftia pachyptila relies entirely on its intracellular chemoautotrophic symbionts to sustain its metabolism. The host must therefore provide them with inorganic metabolites, including carbon. This study describes a tool for studying cell processes occurring in a bacteria-containing cell by the dissociation of trophosome cell types. The physiological assays performed on cell preparations focused on carbon dioxide conversion and transport processes. Trophosome tissue was mechanically dissociated, resulting in cell suspensions enriched in small (7-20 mum) bacteriocytes, which were viable for several hours. In addition, medium-term cell cultures were also attempted. As a start to the understanding of the CO2 metabolism of these cells, we were interested in evidence of carbonic anhydrase (CA) isoforms, ATPases and chloride exchangers. Variations in intracellular and extracellular pH, and in intracellular concentrations of sodium, potassium and chloride, were followed after addition of selective inhibitors. The data presented here suggest the occurrence of potential cytosolic and membrane-associated carbonic anhydrase isoforms in the bacteriocytes, proton-driven sodium-ATPases and a well represented anion transporter exchanging intracellular chloride against extracellular anions.