Inferring the ancestral function of the posterior Hox gene within the bilateria: controlling the maintenance of reproductive structures, the musculature and the nervous system in the acoel flatworm Isodiametra pulchra

Molecular phylogenies place the acoel flatworms as the sister-group to the remaining Bilateria, a position that should prove very valuable when trying to understand the evolutionary origins of the bilaterian body plan. A major feature characterizing Bilateria is the presence of two, orthogonal, body axis. In this article we aim at tackling the problem of how the bilaterian anterior-posterior (AP) axis is organized, and how this axis have been established over evolutionary time. To this purpose we have studied the role of some key regulatory genes involved in the control of the AP axis, the Hox family of transcription factors. All acoels studied to date contain a minimal complement of three Hox genes that are all expressed in nested domains along this major axis, providing the oldest evidence for a "Hox vectorial system" working in Bilateria. However, this proposition is not based in the analysis of Hox functions. Here we document the specific roles of one posterior Hox gene, IpHoxPost, in the postembryonic development of the acoel Isodiametra pulchra. The analysis has been done using RNA interference technologies, for the first time in acoels, and we demonstrate that the functions of this gene are restricted to the posterior region of the animal, within the muscular and neural tissues. We conclude, therefore, that the posterior Hox genes were used to specify and maintain defined anatomical regions within the AP axis of animals since the beginning of bilaterian evolution.