Cedarville University Research and Scholarship Symposium
nociceptin, Tetrahymena thermophila
Tetrahymena thermophila are free-living, ciliated, eukaryotic organisms that respond to stimuli by moving toward chemoattractants and avoiding chemorepellents. Chemoattractant responses involve faster ciliary beating, which propels the organisms forward more rapidly. Chemorepellents signaling involves ciliary reversal, which disrupts forward swimming, and causes the organisms to jerk back and forth, swim in small circles, or spin in an attempt to get away from the repellent. Many food sources, such as proteins, are chemoattractants for these organisms, while a variety of compounds are repellents. Repellents in nature are thought to come from the secretions of predators, or from ruptured organisms, which may serve as "danger" signals. Interestingly, several hormones involved in human pain signaling have been shown to be chemorepellents in Tetrahymena, including substance P, ACTH, PACAP, VIP, and nociceptin.
Recently, we have been studying Tetrahymena response to nociceptin, using pharmacological inhibitors in order to elucidate components of the nociceptin signaling pathway. We have found that G-protein inhibitors and a number of mammalian tyrosine kinase inhibitors have no effect on nociceptin avoidance. However, the tyrosine kinase inhibitor, genistein, inhibits avoidance to nociceptin. Nociceptin avoidance is also inhibited by the calcium chelator, EGTA, which implicates calcium in the avoidance response. Electrophysiology studies done in a calcium-containing buffer show that 50 μM nociceptin causes a sustained depolarization of approximately 30 mV, further supporting the hypothesis that calcium is involved in nociceptin signaling.
J-113397, an inhibitor of the human nociceptin receptor, also inhibits nociceptin avoidance in Tetrahymena. We are currently working to determine whether other inhibitors of the human nociceptin receptor have any effect on Tetrahymena, in order to get a more complete picture of the signaling pathway.
Lampert, T., Bertagnoli, T., Braun, N., Forsberg, J., Gibson, B., Sharon Kobinah, Sarah Stevens, Heather Kuruvilla, (2011). Nociceptin Signals Through Calcium in Tetrahymena thermophila. Cedarville University Research and Scholarship Symposium, Cedarville, Ohio.