29 August 2017

Benson and colleagues, 2017

Benson A, Stadele C, Gahrs C, Vidal-Gadea A, Stein W. 2017. Behavioral consequences of RNA-mediated suppression of innexin expression in marbled crayfish. Illinois State University graduate student symposium. Normal, Illinois, March 2017. https://pdfs.semanticscholar.org/bbd8/0f8501fcf81c0605a61fc6cd1622f983f12f.pdf


We are using a new genetic model system, the marbled crayfish, Procambarus virginalis, for examining the causal relationship between genes, neurophysiology and behavior. Due to their parthenogenetic reproduction, short reproductive life cycle (mature after 2-3 months) and ex-utero breeding, marbled crayfish are ideal for studying the role of genes in producing behavioral output. We are employing RNA interference (RNAi) to suppress gene expression and cell-specific GFP (green fluorescent protein) expression to identify neurons involved in controlling behavior.

Innexins serve as the structural components of gap junctions in invertebrates and build transmembrane channels that mediate electrical coupling between neurons, and facilitate cell-cell communication through rapid movement of ions, electrical impulses, and small messenger molecules. Currently, eight distinct Innexin genes have been found. Despite a high degree of homology between different species, little is known about the role the different Innexins have in shaping behavior.

We use RNAi to suppress Innexin-4 expression in marbled crayfish. We hypothesize that a reduction inInnexin-4 expression leads to an impairment of walking behavior and a deficit in tail-flip escape responses, since both of these behaviors depend on rapid cell-cell communication through gap junctions. Our analysis revealed that the marbled crayfish Innexin-4 shows strong homology to other invertebrate species (e.g. D. melanogaster and C. elegans). We constructed double stranded RNA (dsRNA, ~800 base pairs) containing exonic regions of the Innexin gene. After direct injection of juvenile marbled crayfish with Innexin-4 dsRNA, we are monitoring the animals for several days to evaluate changes in walking behavior and tail-flip escape responses. Our preliminary data suggest that both behaviors are reduced after Innexin4-RNAi.

Keywords: None provided.

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