Clodronate Liposome-Mediated Phagocytic Hemocyte Depletion Affects the Regeneration of the Cephalic Tentacle of the Invasive Snail, <i>Pomacea canaliculata</i>
2023
Giulia Bergamini | Sandro Sacchi | Anita Ferri | Nicola Franchi | Monica Montanari | Mohamad Ahmad | Chiara Losi | Milena Nasi | Marina Cocchi | Davide Malagoli
After amputation, granular hemocytes infiltrate the blastema of regenerating cephalic tentacles of the freshwater snail <i>Pomacea canaliculata</i>. Here, the circulating phagocytic hemocytes were chemically depleted by injecting the snails with clodronate liposomes, and the effects on the cephalic tentacle regeneration onset and on <i>Pc</i>-Hemocyanin, <i>Pc</i>-transglutaminase (<i>Pc</i>-TG) and <i>Pc</i>-Allograft Inflammatory Factor-1 (<i>Pc</i>-AIF-1) gene expressions were investigated. Flow cytometry analysis demonstrated that clodronate liposomes targeted large circulating hemocytes, resulting in a transient decrease in their number. Corresponding with the phagocyte depletion, tentacle regeneration onset was halted, and it resumed at the expected pace when clodronate liposome effects were no longer visible. In addition to the regeneration progress, the expressions of <i>Pc</i>-Hemocyanin, <i>Pc</i>-TG, and <i>Pc-</i>AIF-1, which are markers of hemocyte-mediated functions like oxygen transport and immunity, clotting, and inflammation, were modified. After the injection of clodronate liposomes, a specific computer-assisted image analysis protocol still evidenced the presence of granular hemocytes in the tentacle blastema. This is consistent with reports indicating the large and agranular hemocyte population as the most represented among the professional phagocytes of <i>P. canaliculata</i> and with the hypothesis that different hemocyte morphologies could exert diverse biological functions, as it has been observed in other invertebrates.
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