Repat33 Acts as a Downstream Component of Eicosanoid Signaling Pathway Mediating Immune Responses of Spodoptera exigua, a Lepidopteran Insect

Simple Summary Eicosanoids are oxygenated polyunsaturated fatty acids containing 20 carbons and subdivided into prostaglandin, leukotriene, and epoxyeicosatrienoic acid. They mediate immune responses in insects as well as mammals. Excessive eicosanoids cause severe inflammatory diseases in humans. However, a lack of eicosanoids induces immunosuppressive conditions in insects, which are highly susceptible to various entomopathogens. Despite the physiological significance of eicosanoids, their molecular action was not clearly understood in insects. This study was focused on Repat (Response to Pathogen) gene family, which might be regulated by eicosanoids. Among 44 members of Repat family genes, Repat33 was highly inducible to Gram-negative bacteria. Its expression was dependent on eicosanoids. Loss of function of Repat33 by RNA interference (RNAi) caused significant immunosuppression of a lepidopteran insect, Spodoptera exigua. Larvae of S. exigua treated by RNAi did not exhibit efficient cellular immune responses. They also failed to express antimicrobial peptide genes at high inducible levels in response to bacterial challenges. These results suggest that Repat33 is a downstream component of eicosanoid immune mediation. Repat (=response to pathogen) is proposed for an immune-associated gene family from Spodoptera exigua, a lepidopteran insect. In this gene family, 46 members (Repat1-Repat46) have been identified. They show marked variations in their inducible expression patterns in response to infections by different microbial pathogens. However, their physiological functions in specific immune responses and their interactions with other immune signaling pathways remain unclear. Repat33 is a gene highly inducible by bacterial infections. The objective of this study was to analyze the physiological functions of Repat33 in mediating cellular and humoral immune responses. Results showed that Repat33 was expressed in all developmental stages and induced in immune-associated tissues such as hemocytes and the fat body. RNA interference (RNAi) of Repat33 expression inhibited the hemocyte-spreading behavior which impaired nodule formation of hemocytes against bacterial infections. Such RNAi treatment also down-regulated expression levels of some antimicrobial genes. Interestingly, Repat33 expression was controlled by eicosanoids. Inhibition of eicosanoid biosynthesis by RNAi against a phospholipase A(2) (PLA(2)) gene suppressed Repat33 expression while an addition of arachidonic acid (a catalytic product of PLA(2)) to RNAi treatment recovered such suppression of Repat33 expression. These results suggest that Repat33 is a downstream component of eicosanoids in mediating immune responses of S. exigua.

Automated summary

The gene Repat33 in insects is regulated by eicosanoids, and knocking down its expression can significantly suppress immune responses in Spodoptera exigua through RNA interference.