10.6084/m9.figshare.6200501.v1 David A. Gray David A. Gray Scherezade Hormozi Scherezade Hormozi Fritz R. Libby Fritz R. Libby Randy W. Cohen Randy W. Cohen Supplementary Methods and Figures S1 - S4 from Induced expression of a vestigial sexual signal The Royal Society 2018 vestigial behaviour sexual signals Gryllus 2018-04-30 12:14:58 Journal contribution https://rs.figshare.com/articles/journal_contribution/Supplementary_Methods_and_Figures_S1_-_S4_from_Induced_expression_of_a_vestigial_sexual_signal/6200501 Vestigial morphological traits are common and well known in a variety of taxa. Identification of vestigial genes has illustrated the potential for evolutionary reversals and the re-expression of atavistic traits. Here we induce expression of a behavioural sexual signal, male calling song, in a cricket species, <i>Gryllus ovisopis,</i> which lacks a functional calling song. We successfully used acetylcholine injections in the frontal space of the head of male crickets to activate cerebral command neurons for cricket calling, and we recorded calling songs with a temporal chirp pattern similar to that of <i>G. ovisopis</i>' close evolutionary relatives, <i>G. firmus</i> and <i>G. pennsylvanicus</i>, implying that the neural pattern generators which underlie cricket calling behaviour persist in a vestigial state in <i>G. ovisopis</i>. To our knowledge, this is the first demonstration of the induced expression of a vestigial behaviour in any organism. The retention of latent neural capacity to express sexual behaviours could have important implications for rapid evolution, trait re-emergence and reproductive isolation.