Abstract

Here we report the characterization of an octopamine/tyramine (OA/TA or TyrR1) receptor (OA/TAMac) cloned from the freshwater prawn, Macrobrachium rosenbergii, an animal used in the study of agonistic social behavior. The invertebrate OA/TA receptors are seven trans-membrane domain G-protein coupled receptors that are related to vertebrate adrenergic receptors. Behavioral studies in arthropods indicate that octopaminergic signaling systems modulate fight or flight behaviors with octopamine and/or tyramine functioning in a similar way to the adrenalins in vertebrate systems. Despite the importance of octopamine signaling in behavioral studies of decapod crustaceans there are no functional data available for any of their octopamine or tyramine receptors. We expressed OA/TAMac in Xenopus oocytes where agonist-evoked trans-membrane currents were used as readouts of receptor activity. The currents were most effectively evoked by tyramine but were also evoked by octopamine and dopamine. They were effectively blocked by yohimbine. The electrophysiological approach we used enabled the continuous observation of complex dynamics over time. Using voltage steps, we were able to simultaneously resolve two types of endogenous currents that are affected over different time scales. At higher concentrations we observe that octopamine and tyramine can produce different and opposing effects on both of these currents, presumably through the activity of the single expressed receptor type. The pharmacological profile and apparent functional-selectivity are consistent with properties first observed in the OA/TA receptor from the insect Drosophila melanogaster. As the first functional data reported for any crustacean OA/TA receptor, these results suggest that functional-selectivity between tyramine and octopamine is a feature of this receptor type that may be conserved among arthropods.

Highlights

  • Octopamine, tyramine, and dopamine are structurally similar biogenic amines that are derived from the amino acid tyrosine (Fig. 1) [1]

  • We are currently cloning and characterizing aminergic G protein-coupled receptors (GPCRs) from the giant tropical prawn Macrobrachium rosenbergii as a step toward elucidating the molecular mechanisms associated with the formation and maintenance of prawn social hierarchies [14,15]

  • In addition to functional selectivity evoked through different agonists, concentration-sensitive effects caused by high and low concentrations of the same agonist have been observed for a number of insect octopamine receptors [27,30,44]

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Summary

Introduction

Octopamine, tyramine, and dopamine are structurally similar biogenic amines that are derived from the amino acid tyrosine (Fig. 1) [1]. In addition to functional selectivity evoked through different agonists, concentration-sensitive effects caused by high and low concentrations of the same agonist have been observed for a number of insect octopamine receptors [27,30,44]. At high concentrations, tyramine increases, whereas octopamine decreases ICl-T This is the first report of opposing cellular effects between tyramine and octopamine observed following the expression of an OA/TA receptor. This finding on the crustacean receptor agrees with previous findings from the insect OA/TA receptor (CG7485) that suggest the apparently minor chemical modification (a single hydroxyl group) between tyramine and octopamine (Fig. 1) can cause altered function of the OA/TA receptor [43]. These data provide evidence that functional selectivity between octopamine and tyramine may be a conserved property of arthropod OA/TA receptors

Results and Discussion
Materials and Methods
Methods
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