Transcriptional Fusions of Putative G‐protein Coupled‐Receptors from Hookworm (Ancylostoma ceylanicum) Expressed in Caenorhabditis elegans

Abstract

Many parasites, including hookworms of the genus Ancylostoma, are specialists and require host‐specific signals to resume development upon infection. Although little is known about the nature of these signaling molecules, evolutionary conservation between hookworms and the model nematode C. elegans suggests that hookworm host‐signal receptors are likely to be G‐protein coupled receptors (GPCRs) that are expressed in amphid neurons. Based on bioinformatic analysis of developmental stage‐specific RNA‐Seq data from A. caninum and A. ceylanicum, predicted hookworm GPCRs with increased expression levels during the infective larval stage 3 (iL3) were identified.Approximately 1500 bp of genomic sequence upstream of the predicted start site for the putative A. ceylanicum GPCRs were fused to green fluorescent protein (GFP) coding sequence to generate promoter‐GFP transcriptional fusion constructs, which were microinjected into C. elegans. Of nine hookworm promoter fusions tested, four drove differential GFP expression in C. elegans tissues, including the digestive tract, pharynx, and a neuron pair. Preliminary analysis identified the neurons as interneurons, potentially involved in locomotion responses to chemosensory input in C. elegans. The putative GPCR encoded by the neuronally expressed gene is upregulated in both A. ceylanicum and A. caninum, suggesting that it may play a general role in hookworm development during host infection. These experiments indicate that hookworm promoters can be expressed in C. elegans, providing a surrogate system for the study of hookworm gene expression patterns.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.