Investigation of the Neuronal Expression Pattern of the G Protein-coupled Receptor FSHR-1 in C. elegans

Emily Nettesheim Butler University
Faculty Sponsor(s): Jennifer Kowalski Butler University
The nervous system relies on numerous proteins to regulate neural signaling. FSHR-1 is a G protein-coupled receptor in Caenorhabditis elegans and a homolog of mammalian glycopeptide hormone receptors. C. elegans FSHR-1 regulates immune responses, germline differentiation, and muscle contraction, but FSHR-1 neuronal functions and site(s) of action are not fully characterized. This project aims to define the neurons in which FSHR-1 is expressed in the model roundworm C. elegans. Our unpublished data suggest FSHR-1 can act in both excitatory and inhibitory motor neurons to control neuromuscular signaling; thus, we hypothesize fshr-1 is normally expressed in inhibitory GABAergic and/or excitatory cholinergic neurons. Imaging experiments using strains expressing the transcriptional reporter Pfshr-1::gfp and a separate fluorescent marker driven under a promoter for a specific neuronal subclass (sensory, GABAergic, cholinergic, or glutamatergic neurons) showed little colocalization of fshr-1 with these markers. This may be due to missing regulatory elements within the four kilobase promoter used in these studies, as such elements may be at distant chromosomal sites. We are now using the clustered regularly-interspaced short palindromic repeats (CRISPR)/Cas9 system to tag endogenous fshr-1 with gfp in order to determine the complete set of cells in which endogenous fshr-1 is expressed. Dokshin et al (2018) recently introduced a new C. elegans CRISPR method that uses partially single-stranded repair templates. Experiments with this protocol are ongoing. Determining the cells where fshr-1 is expressed is critical for understanding FSHR-1’s role in neuronal signaling, which may be important in understanding and treating neurological disorders involving signaling imbalances.
Biology
Oral Presentation

When & Where

11:00 AM
Gallahue Hall 105