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Toll/Toll-like receptors are key components in the innate immune responses of invertebrates. In this study, we identified two novel Toll receptors ( PcToll5  and  PcToll6 ) from the red swamp crayfish  Procambarus clarkii . The complete cDNA sequence of  PcToll5  is 4247 bp, encoding a 1293 amino acid polypeptide. The full-length 4688 bp  PcToll6  encodes a putative protein of 1195 amino acids. Quantitative RT-PCR analysis indicated that  PcToll5  and  PcToll6  were constitutively expressed in all tissues studied. The highest expression levels of  PcToll5  and  PcToll6  were found in the intestine and gills, respectively, and were significantly upregulated from 24 to 48 h during white spot syndrome virus (WSSV) challenge. siRNA-mediated RNA interference results showed that  PcToll5  and  PcToll6  might regulate the expression of anti-lipopolysaccharide factors ( PcALF2  and  PcALF3 )  in vivo . Overexpression of PcToll5 and PcToll6 in  Drosophila  Schneider 2 (S2) cells activated the transcription of  Drosophila  antimicrobial peptides, including  drosomycin  ( Drs ),  metchnikowin  ( Mtk ), and  attacin A  ( AttA ), and shrimp  Penaeidin -4 ( Pen4 ). These findings provide significant information that  PcToll5  and  PcToll6  may contribute to host immune defense against WSSV in  P. clarkii .

frontiers in physiology

Cloning and Characterization of Two Toll Receptors (PcToll5 and PcToll6) in Response to White Spot Syndrome Virus in the Red Swamp Crayfish Procambarus clarkii