Patients treated with amiodarone accumulate lysobisphosphatidic acid (LBPA), also known as bis(monoacylglycero)phosphate, in airway secretions and develop in different tissues vacuoles and inclusion bodies thought to originate from endosomes. To clarify the origin of these changes, we studied in vitro the effects of amiodarone on endosomal activities like transferrin recycling, Shiga toxin processing, ESCRT-dependent lentivirus budding, fluid phase endocytosis, proteolysis and exosome secretion. Furthermore, since the accumulation of LBPA might point to a broader disturbance in lipid homeostasis, we studied the effect of amiodarone on the distribution of LBPA, unesterified cholesterol, sphingomyelin and glycosphyngolipids. Amiodarone analogues were also studied, including the recently developed derivative dronedarone. We found that amiodarone does not affect early endosomal activities, like transferrin recycling, Shiga toxin processing and lentivirus budding. Amiodarone, instead, interferes with late compartments of the endocytic pathway, blocking the progression of fluid phase endocytosis and causing fusion of organelles, collapse of lumenal structures, accumulation of undegraded substrates and amassing of different types of lipids. Not all late endocytic compartments are affected, since exosome secretion is spared. These changes recall the Niemann-Pick type-C phenotype (NPC), but originate by a different mechanism, since, differently from NPC, they are not alleviated by cholesterol removal. Studies with analogues indicate that basic pKa and high water-solubility at acidic pH are crucial requirements for the interference with late endosomes/lysosomes and that, in this respect, dronedarone is at least as potent as amiodarone. These findings may have relevance in fields unrelated to rhythm control.

The expression of alpha-gustducin, a G protein alpha subunit involved in bitter and sweet taste transduction, was investigated in chemosensory tissues of adult mice. By immunohistochemistry, alpha gustducin was absent in the olfactory neuroepithelium. Instead, alpha gustducin was expressed in a subset of bipolar cells in the proliferative zone of the vomeronasal neuroepithelium as well as in taste buds. Northern blot analysis confirmed the presence of alpha gustducin in isolated vomeronasal organs. Moreover, immunohisto- chemistry revealed the expression of alpha gustducin in scattered cells of the nasal respiratory epithelium. These results show for the first time that alpha gustducin is expressed in chemosensory tissue outside the alimentary tract, suggesting that common transduction mechanisms could be shared by apparently unrelated chemosensory tissues.

The vomeronasal organ of frog and mouse was investigated for the presence and content of serotonin and catecholamines by means of high-performance liquid chromatography. Measurable amounts of serotonin, adrenaline and noradrenaline were found in the vomeronasal organ of adult individuals of both species. The amine content varied with sex of adult frogs and mice and sexual maturity of mice. In preliminary experiments, acute exposure to male urine containing pheromone affected the amine content in the vomeronasal organ of adult female mice. These data suggest that functional sex dimorphism is present in the vomeronasal organ, and biochemical changes therein take place according to stage of sexual maturity. The role of biogenic amines in the vomeronasal organ deserves further study.

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