Phenotypic variation reveals sites of evolutionary constraint in the androgenic signaling pathway

Steroid hormone systems play an important role in shaping the evolution of vertebrate sexual traits, but several aspects of this relationship remain unclear. For example, we currently know little about how steroid signaling complexes are adapted to accommodate the emergence of behavior in response to sexual selection. We use downy woodpeckers (Dryobates pubescens) to evaluate how the machinery underlying androgen action can evolve to accommodate this bird's main territorial signal, the drum. We focus specifically on modifications to androgenic mechanisms in the primary neck muscle that actuates the hammering movements underlying this signal. Of the signaling components we examine, we find that levels of circulating testosterone (T) and androgen receptor (AR) expression are consistently increased in a way that likely enhances androgenic regulation of drumming. By contrast, the expression of nuclear receptor co-factors-the 'molecular rheostats' of steroid action-show no such relationship in our analyses. If anything, co-factors are expressed in directions that would presumably hinder androgenic regulation of the drum. These findings therefore collectively point to T levels and AR as the more evolutionarily labile components of the androgenic system, in that they are likely more apt to change over time to support sexual selection for territorial signaling in woodpeckers. Yet the signaling elements that fine-tune AR's functional effects on the genome-namely the receptor's transcriptional co-factors-do not change in such a manner, and thus may be under tighter evolutionary constraint.