Adachi, M. | Search bioRxiv

bioRxiv2021

Green spotted puffer can detect an almost nontoxic TTX analog odor using crypt olfactory sensory neurons

Tetrodotoxin (TTX) is a well-known neurotoxin that functions as a defense substance for toxic puffers. Several behavioral studies reported that TTX attracts toxic puffers belonging to the genus Takifugu. Although our electrophysiological and behavioral studies showed that a TTX analog, 5,6,11-trideoxyTTX, acts as an olfactory chemoattractant for grass puffers (T. alboplumbeus), it is unclear whether toxic puffers are commonly attracted to 5,6,11-trideoxyTTX, and which types of olfactory sensory neurons (OSNs) detect 5,6,11-trideoxyTTX. Here we investigated whether the green spotted puffer (Dichotomyctere nigroviridis), a phylogenetically different species from the grass puffer, is attracted to 5,6,11-trideoxyTTX. Administration of 5,6,11-trideoxyTTX attracted green spotted puffers, but TTX or vehicle did not. Furthermore, immunohistochemistry of the olfactory epithelium exposed to 5,6,11-trideoxyTTX with an antibody against phosphorylated ribosomal protein S6 (pS6), a neuronal activity marker, labeled oval cells with apical invagination. These oval cells were also labeled by the antibody against S100, a specific marker of crypt OSNs. Thus, our results suggest that 5,6,11-trideoxyTTX acts as an olfactory chemoattractant that is detected by crypt-type OSNs in the olfactory epithelium of green spotted puffers. Toxic puffers may use 5,6,11-trideoxyTTX as an olfactory chemoattractant involved in reproduction and parental care or as an olfactory cue of TTX-bearing organisms for effective toxification. Summary statementBehavioral and immunohistochemical experiments suggest that an almost nontoxic TTX analog, 5,6,11-trideoxyTTX, acts as an olfactory chemoattractant for green spotted puffers, and crypt-type olfactory sensory neurons detect it.

zoology↗

bioRxiv2021

Almost nontoxic tetrodotoxin analog, 5,6,11-trideoxytetrodotoxin, as an olfactory chemoattractant for the grass puffer

Toxic puffers contain the potent neurotoxin, tetrodotoxin (TTX). Although TTX is considered to serve as a defense substance, previous behavioral studies have demonstrated that TTX acts as an attractive pheromone for some toxic puffers. To elucidate the physiological mechanism of putative pheromonal action of TTX, we examined whether grass puffers Takifugu alboplumbeus can detect TTX. Electroolfactogram (EOG) results suggest that the olfactory epithelium (OE) of grass puffers responded to a type of TTX analog (5,6,11-trideoxyTTX), although it did not respond to TTX. We also examined the attractive action of 5,6,11-trideoxyTTX on grass puffers by recording their swimming behavior under dark conditions. Grass puffers preferred to stay on the side of the aquarium where 5,6,11-trideoxyTTX was administered, and their swimming speed decreased. Additionally, odorant-induced labeling of olfactory sensory neurons by immunohistochemistry against neural activity marker (phosphorylated extracellular signal regulated kinase; pERK) revealed that labeled olfactory sensory neurons were localized in the region surrounding "islets" where there was considered as nonsensory epithelium. 5,6,11-trideoxyTTX has been known to accumulate in grass puffers, but its toxicity is much lower (almost nontoxic) than TTX. Our results suggest that toxic puffers may positively use this TTX analog, which has been present in their body with TTX but whose function was unknown, as an odorant for chemical communication or effective TTX accumulation.

zoology↗