Perfused Gills Reveal Fundamental Principles of pH Regulation and Ammonia Homeostasis in the Cephalopod Octopus vulgaris

Marian Y. Hu, Po-Hsuan Sung, Ying-Jey Guh, Jay-Ron Lee, Pung-Pung Hwang, Dirk Weihrauch, Yung-Che Tseng

In contrast to terrestrial animals most aquatic species can be characterized by relatively higher blood [Formula: see text] concentrations despite its potential toxicity to the central nervous system. Although many aquatic species excrete [Formula: see text] via specialized epithelia little information is available regarding the mechanistic basis for NH3/[Formula: see text] homeostasis in molluscs. Using perfused gills of Octopus vulgaris we studied acid-base regulation and ammonia excretion pathways in this cephalopod species. The octopus gill is capable of regulating ammonia (NH3/[Formula: see text]) homeostasis by the accumulation of ammonia at low blood levels (<260 μM) and secretion at blood ammonia concentrations exceeding in vivo levels of 300 μM. [Formula: see text] transport is sensitive to the adenylyl cyclase inhibitor KH7 indicating that this process is mediated through cAMP-dependent pathways. The perfused octopus gill has substantial pH regulatory abilities during an acidosis, accompanied by an increased secretion of [Formula: see text]. Immunohistochemical and qPCR analyses revealed tissue specific expression and localization of Na(+)/K(+)-ATPase, V-type H(+)-ATPase, Na(+)/H(+)-exchanger 3, and Rhesus protein in the gill. Using the octopus gill as a molluscan model, our results highlight the coupling of acid-base regulation and nitrogen excretion, which may represent a conserved pH regulatory mechanism across many marine taxa.