Side-by-side secretion of Late Palaeozoic diverged courtship pheromones in an aquatic salamander

Ines Van Bocxlaer, Dag Treer, Margo Maex, Wim Vandebergh, Sunita Janssenswillen, Gwij Stegen, Philippe Kok, Bert Willaert, Severine Matthijs, Erik Martens, Anneleen Mortier, Henri De Greve, Paul Proost, Franky Bossuyt

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Males of the advanced salamanders (Salamandroidea) attain internal fertilization without a copulatory organ by depositing a spermatophore on the substrate in the environment, which females subsequently take up with their cloaca. The aquatically reproducing modern Eurasian newts (Salamandridae) have taken this to extremes, because most species do not display close physical contact during courtship, but instead largely rely on females following the male track at spermatophore deposition. Although pheromones have been widely assumed to represent an important aspect of male courtship, molecules able to induce the female following behaviour that is the prelude for successful insemination have not yet been identified. Here, we show that uncleaved sodefrin precursor-like factor (SPF) protein pheromones are sufficient to elicit such behaviour in female palmate newts (Lissotriton helveticus). Combined transcriptomic and proteomic evidence shows that males simultaneously tail-fan multiple ca 20 kDa glycosylated SPF proteins during courtship. Notably, molecular dating estimates show that the diversification of these proteins already started in the late Palaeozoic, about 300 million years ago. Our study thus not only extends the use of uncleaved SPF proteins outside terrestrially reproducing plethodontid salamanders, but also reveals one of the oldest vertebrate pheromone systems.
Original languageEnglish
Article number20142960
Number of pages9
JournalProceedings of the Royal Society B: Biological Sciences
Issue number1803
Early online date18 Feb 2015
Publication statusPublished - Mar 2015


  • Amphibians
  • Evolution
  • Gene Duplications
  • Phylogeny
  • Protein Pheromones


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