Isolation and transcriptome analysis of three subpopulations of shrimp hemocytes reveals the underlying mechanism of their immune functions

Abstract

Hemocytes in shrimp play important roles in innate immune responses against pathogens. Although three types of hemocytes including hyalinocytes, semi-granulocytes and granulocytes were identified based on their morphological characters in penaeid shrimp, knowledge about the molecular basis of their functions in the immunity is still very limited. In the present study, three subpopulations of hemocytes were firstly separated by Percoll gradient centrifugation, and their transcriptomes were analyzed. The data showed that significantly differential gene expression patterns existed in different types of hemocytes. The genes encoding phagocytic receptors, lectins and actin cytoskeleton involved in phagocytosis were highly expressed in hyalinocytes, while genes involved in the humoral immunity signaling pathways were highly expressed in semi-granulocytes, and genes encoding prophenoloxidase (proPO)-activating enzyme and serine proteases involved in proPO system activation were highly expressed in granulocytes. Further flow cytometry analysis indicated that hyalinocytes were the main hemocytes subpopulation responsible for ingesting foreign fluorescent beads, and this ingestion process mainly depends on the endocytic way of macropinocytosis. These data provide valuable information for understanding the molecular basis of distinct shrimp hemocytes subpopulations of shrimp in cellular and humoral immunity.