PGE2 mediates hemocyte-spreading behavior by activating aquaporin via cAMP and rearranging actin cytoskeleton via Ca2

Abstract

Spreading behavior of hemocytes (= insect blood cells) is essential for cellular immune responses against various microbial pathogens. It is activated by prostaglandin E₂ (PGE₂) via its membrane receptor associated with secondary messenger, cAMP, in insects. This study observed an increase of calcium ion (Ca²⁺) level after an acute increase of cAMP induced by PGE₂ treatment and clarified the intracellular signals underlying the hemocyte-spreading behavior. Inhibition of Ca²⁺ flux significantly impaired the hemocyte-spreading and subsequent cellular immune response, phagocytosis. The up-regulation of intracellular Ca²⁺ in response to PGE₂ was dependent on cAMP because RNA interference (RNAi) of PGE₂ receptor expression or inhibiting adenylate cyclase prevented Ca²⁺ mobilization. The up-regulation of Ca²⁺ was induced by inositol triphosphate (IP₃) via its specific IP₃ receptor. Furthermore, inhibition of ryanodine receptor impaired Ca²⁺ mobilization, suggesting Ca²⁺-induced Ca²⁺ release. However, the effective spreading behavior of hemocytes was dependent on both secondary messengers. Ca²⁺ signal stimulated by cAMP was required for activating small G proteins because RNAi treatments of small G proteins such as Rac1, RhoA, and Cdc42 failed to stimulate hemocyte-spreading. In contrast, aquaporin was activated by cAMP. Its activity was necessary for changing cell volume during hemocyte-spreading. These results indicate that PGE₂ mediates hemocyte-spreading via cAMP signal to activate aquaporin and via Ca²⁺ signal to activate actin cytoskeletal rearrangement.

Keywords: Aquaporin; Calcium; Hemocyte; Immunity; PGE(2); cAMP.