Mixture suppression without inhibition for binary mixtures from whole cell patch clamp studies of in situ olfactory receptor neurons of the spiny lobster

Abstract

Whole cell patch clamping was used to investigate mechanisms of mixture suppression for in situ olfactory receptor neurons (ORNs) of the spiny lobster Panulirus argus. We used a set of single compounds and binary mixtures that have been used in previous biochemical studies of receptor-odorant binding, electrophysiological studies of spiking output from ORNs, and behavioral studies. These odorants were adenosine 5′-monophosphate (AMP), betaine (Bet), l-cysteine (Cys), l-glutamate (Glu), taurine (Tau), ammonium chloride, d,l-succinate, binary mixtures of these compounds, as well as a 33-component artificial oyster mixture (AOM). For the 40 ORNs studied, these stimuli more frequently elicited inward than outward currents. AMP, Glu, Tau and Bet evoked the largest and most numerous inward currents; Cys most commonly evoked outward currents. Na + was an important charge-carrying ion for the Glu-evoked response in one ORN and the Bet-evoked response in another ORN. Mixture suppression, defined conservatively in this study as cases where the response to a binary mixture was less than the response to the more excitatory component of that mixture, was observed in 6 ORNs. In all 6 cases, neither component of the mixture evoked an outward conductance (i.e. neither was inhibitory). Five of these cases of mixture suppression involved a mixture containing two excitatory compounds (i.e. producing inward conductances): four ORNs were excited by both Glu and AMP, and one ORN was excited by both Tau and Glu. One case of mixture suppression occurred for a compound (Tau) that did not produce a current when presented alone but which when added to Bet suppressed the inward current generated by Bet. Mechanisms for these suppressions are discussed, including inhibition of receptor binding by the components of a binary mixture and effects on second messengers or ion channels.