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

In contrast to terrestrial animals most aquatic species can be characterized by relatively higher blood NH4+ concentrations despite its potential toxicity to the central nervous system. Although many aquatic species excrete NH4+ via specialized epithelia little information is available regarding the mechanistic basis for NH3/NH4+ 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/NH4+) 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. NH4+ 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 NH4+. 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.