MSc Exit Seminar-Thomas Dodsworth

Characterization of the Teneurin C-terminal Associated Peptide (TCAP) and Latrophilin Ligand-Receptor Pair in an Immortalized Skeletal Muscle Cell Line

Teneurin C-terminal associated peptide (TCAP) is an ancient and conserved bioactive peptide that is evolutionarily related to corticotropin releasing factor (CRF). Recently, synthetic TCAP-1 was shown to increase cellular energy availability and alter contractile performance in rodent skeletal muscle. However, the exact receptor signalling mechanism through which this occurs is unknown. Based on evidence of their interaction in vitro, I hypothesized that TCAP-1 signals through latrophilins—a family of adhesion G-protein coupled receptors—to elicit its effects in muscle. To test this, I knocked-down and knocked-out latrophilins in the immortalized mouse myoblast C2C12 cell line by small interfering RNA (siRNA) and CRISPR/Cas9 gene editing methods, respectively, and examined the efficacy of TCAP-1 in knockdown and knockout cells. I determined that latrophilin-1 is necessary for TCAP-1-mediated increases in intracellular calcium, NADH turnover, and PGC-1a expression. This establishes, for the first time, that the TCAP-latrophilin ligand-receptor pair has a functional role in skeletal muscle.