Novel Neurotoxic Activity in Calliophis intestinalis Venom

In this work, we investigated the in vitro neurotoxicity of Calliophis intestinalis venom using chick biventer cervicis neuromuscular preparations and electrophysiological analysis of voltage-gated sodium (Na_V) channels expressed in HEK293 cells. We found that the indirect twitches of the neuromuscular preparations decreased over time when exposed to venom. However, the responses of these preparations to the agonists acetylcholine, carbachol, and potassium chloride were not changed after incubation with the venom. Our electrophysiological experiments show that C. intestinalis venom acts as a Na_V channel antagonist-the first known from a vertebrate venom-by decreasing the peak current of Na_V1.4 channels without changing the kinetics of activation or inactivation. Our proteomic results accord with earlier analyses and find that the venom contains three-finger toxins, cysteine-rich secretory proteins, kunitz peptides, phospholipase A₂s, snake venom metalloproteases, and vespryns. Some of the three-finger toxins are similar to the δ-elapitoxins from the venom of the closely related Calliophis bivirgatus. However, δ-elapitoxins act as Na_V channel agonists in C. bivirgatus whereas C. intestinalis venom contains Na_V channel antagonists. The toxins and mechanisms responsible for the neuromuscular symptoms remain unclear as does the identity of the Na_V channel antagonists. These aspects of this unusual venom require further study.