title: The β-blocker propranolol affects cAMP-dependent signaling and induces the stress response in Mediterranean mussels, Mytilus galloprovincialis.
abstract: Widespread occurrence of pharmaceuticals is reported in aquatic systems, posing concerns for the health of aquatic wildlife and a theoretical risk to humans. A recent concept was developed for the identification of highly active compounds amongst the environmental pharmaceuticals, based on their mode of action, the homology between human targets and possible targets in the environment, and the importance of the affected pathway for the target species. In line with this approach, this study investigated whether propranolol (PROP) affects the cAMP-dependent pathway in Mediterranean mussels, Mytilus galloprovincialis. PROP is a prototypical β-adrenoceptor antagonist, and these receptors exist in bivalves and show gross pharmacological properties similar to their mammalian counterparts. PROP also acts as a 5-HT1 receptor antagonist, which is the sole 5-HT receptor reported in bivalves to date. Importantly, β-adrenoceptor and 5HT-1 receptor subtypes are positively and negatively coupled to cAMP-mediated signaling, respectively. PROP was administered as either l-PROP or dl-PROP. A wide range of concentrations was tested including low (0.3, 3 and 30ng/L) and high (300ng/L) environmental ranges, and a concentration 5-fold above the maximum reported environmental level (30,000ng/L). After a 7-day exposure, mussel cAMP levels and PKA activities were significantly reduced in digestive gland, increased in mantle/gonads and unaffected in gills. Similar patterns were observed for the mRNA expression of the ABCB1 gene encoding the membrane transporter P-glycoprotein, hypothesised to be under PKA modulation. The effects on the digestive gland are consistent with PROP blocking β-adrenoceptors. The observed increased cAMP levels in the mantle/gonad tissue support PROP blocking 5-HT1 receptors. Catalase and glutathione-S tranferase were differently affected by PROP in the two tissues. Mussel haemocyte lysosome membrane stability, a sensitive biomarker of animal health status, was concentration-dependently reduced following PROP exposure. Our observations provide evidence for PROP affecting cell signaling in M. galloprovincialis. Moreover, the chemical interacts with specific and evolutionally conserved biochemical pathways for which it was designed. The mode of action of PROP in mussels is related with its therapeutic properties in humans, based upon these conserved human targets. It also induced a stress response, and all these effects were displayed at the lowest concentrations tested.
authors.affiliation: University of Bologna, Interdepartment Centre for Research in Environmental Sciences (CIRSA), via S. Alberto 163, 48123 Ravenna, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Franzellitti
authors.forename: Sara
authors.initials: S
authors.lastname: Buratti
authors.forename: Paola
authors.initials: P
authors.lastname: Valbonesi
authors.forename: Antonio
authors.initials: A
authors.lastname: Capuzzo
authors.forename: Elena
authors.initials: E
authors.lastname: Fabbri
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D000818:Animals; D015415:Biomarkers; D002374:Catalase / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D004064:Digestive System / Q000378:metabolism; D005880:Gills / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D006066:Gonads / Q000378:metabolism; D006434:Hemocytes / Q000378:metabolism; D008247:Lysosomes / Q000187:drug effects; D049878:Mytilus / Q000187:drug effects* / Q000378:metabolism; D011433:Propranolol / Q000008:administration & dosage / Q000378:metabolism / Q000494:pharmacology*; D011943:Receptors, Adrenergic, beta / Q000187:drug effects; D014874:Water Pollutants, Chemical / Q000008:administration & dosage / Q000378:metabolism / Q000493:pharmacokinetics
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D015415:Biomarkers; D011943:Receptors, Adrenergic, beta; D014874:Water Pollutants, Chemical; D011433:Propranolol; D000242:Cyclic AMP; D002374:Catalase; D005982:Glutathione Transferase; D017868:Cyclic AMP-Dependent Protein Kinases