Are Pacific Chorus Frogs (Pseudacris regilla) Resistant to Tetrodotoxin (TTX)? Characterizing Potential TTX Exposure and Resistance in an Ecological Associate of Pacific Newts (Taricha)

Animals that frequently encounter toxins often develop mechanisms of toxin resistance over evolutionary time. Both predators that consume toxic prey and organisms in physical contact with a toxin in their environment may experience natural selection for resistance. Based on observations that Pacific Chorus Frogs (Pseudacris regilla) sometimes eat and mistakenly amplect tetrodotoxin (TTX)-defended Taricha newts, we predicted that P. regilla may possess TTX resistance. We compared amino acid sequences of domain IV of the muscle voltage-gated sodium channel gene SCN4A (Na(V)1.4) in populations of P. regilla that are sympatric and allopatric with Taricha. We identified a single substitution in Na(V)1.4 of P. regilla at a conserved site in the pore loop where TTX binds. Although the role of this site in TTX resistance has not been functionally assessed, both allopatric and sympatric P. regilla had this substitution, along with several other reptiles and amphibians, suggesting that it may be unrelated to TTX exposure from Taricha. Thus, there is no conclusive evidence that P. regilla possesses TTX resistance encoded by amino acid substitutions in this domain. California occurrence data from the last 50 yr indicate that Taricha activity peaks in January while the activity of P. regilla peaks in April, with times where the species may come into contact. However, P. regilla may not be exposed to levels of TTX from Taricha high enough to select for mutations in Na(V)1.4. Other unidentified mechanisms of TTX resistance could be present in P. regilla and other species sympatric with toxic newts.