In contrast to the single sensory surface present in teleost fishes, several spatially segregated subsystems with distinct molecular and functional characteristics define the mammalian olfactory system. However, the evolutionary steps of that transition remain unknown. Here we analyzed the olfactory system of an early diverging tetrapod, the amphibian Xenopus laevis, and report for the first time the existence of two odor-processing streams, sharply segregated in the main olfactory bulb and partially segregated in the olfactory epithelium of pre-metamorphic larvae. A lateral odor-processing stream is formed by microvillous receptor neurons and is characterized by amino acid responses and Gαo/Gαi as probable signal transducers, whereas a medial stream formed by ciliated receptor neurons is characterized by responses to alcohols, aldehydes, and ketones, and Gαolf/cAMP as probable signal transducers. To reveal candidates for the olfactory receptors underlying these two streams, the spatial distribution of 12 genes from four olfactory receptor gene families was determined. Several class II and some class I odorant receptors (ORs) mimic the spatial distribution observed for the medial stream, whereas a trace amine-associated receptor closely parallels the spatial pattern of the lateral odor-processing stream. Other olfactory receptors (some class I odorant receptors and vomeronasal type 1 receptors) and odor responses (to bile acids, amines) were not lateralized, the latter not even in the olfactory bulb, suggesting an incomplete segregation. Thus, the olfactory system of X. laevis exhibits an intermediate stage of segregation and as such appears well suited to investigate the molecular driving forces behind olfactory regionalization.

The main olfactory system of larval Xenopus laevis is made up of at least two subsystems consisting of subsets of olfactory receptor neurons (ORNs) with different transduction mechanisms. One ORN subset lacks the canonical cAMP transduction pathway and responds to amino acid odorants. The second subset has the cAMP transduction pathway but as yet suitable odorants are unknown. Here we report the identification of amines as proper olfactory stimuli for larval X. laevis using functional Ca(2+) imaging and slice preparations of the olfactory system. The response profiles of individual ORNs to a number of amines were extremely complex and mostly highly specific. The great majority of amine-sensitive ORNs responded also to forskolin, an activator of the olfactory cAMP transduction pathway. Most amine-induced responses could be attenuated by the cyclic nucleotide-gated channel inhibitor LY83583. This confirms that most amine-responsive olfactory receptors (ORs) are coupled to the cAMP-dependent transduction pathway. Furthermore, we show that trace amine-associated receptors (TAARs), which have been shown to act as specific ORs for amines in mammals, are expressed in the olfactory organ of X. laevis. The TAARs expressed in Xenopus cannot, however, explain the complex responses of individual ORNs to amines because there are too few of them. This indicates that, in addition to TAARs, there must be other receptor families involved in the detection of amines.

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