Serotonin- and eicosanoid-dependent rapid hemocyte aggregation in the hemolymph is the first step in nodule formation in Bombyx mori larvae

[Display omitted] •Hemocyte aggregation played a major role in rapid clearance of invading microorganisms.•Single yeast cell induced hemocyte aggregation suggests that nodule formation is a natural system.•Serotonin and prostaglandin E2 induces hemocyte aggregation accompanied by hemocytin secretion. Nodule formation is a well-known process in cellular immunity of insects. However, few studies have investigated the role of hemocytes in rapid aggregation before tissue adhesion. In addition, since nodule formation is usually elicited by injecting large quantities of microbes, it remains unclear whether nodule formation is a natural response. The present study addressed these issues. A small number of nodules adhered to the dorsal vessels 1 min after Saccharomyces cerevisiae injection, while numerous aggregates of hemocytes and S. cerevisiae cells were observed in the hemolymph. The aggregate number decreased sharply after 5 min, corresponding to a rapid increase in the number of nodules. This suggests that aggregates formed in the hemolymph in response to S. cerevisiae injection eventually attached to the tissues. Nodules were induced using conditions that do not occur in nature, i.e., injection of nearly 2,000,000 S. cerevisiae cells. However, many aggregates contained only one S. cerevisiae cell, suggesting that aggregate formation can begin with the invasion of a single cell and that nodule formation is not an unnatural response. Biosynthesis inhibitors of serotonin (5-HT) and eicosanoids inhibited aggregate and nodule formation. In addition, injection of 5-HT and prostaglandin E2 induced hemocyte aggregation within 1 min in the hemolymph, along with hemocytin release. This suggested that 5-HT and eicosanoids induce rapid aggregation in response to invading microorganisms.