Double abdomen in a short-germ insect: Zygotic control of axis formation revealed in the beetle Tribolium castaneum

Significance One of the first crucial steps of animal development is to distinguish the anterior versus the posterior pole of the embryo, i.e., the AP axis. If this process fails, embryos may develop two mirror image tails or heads. In the fly Drosophila, the mother provides the signals required for AP axis formation, while in vertebrates, gene activity of the embryo is required as well. We identified two genes whose knockdown leads to double-tail phenotypes in the beetle Tribolium, representing the insect-typical short-germ embryogenesis. Intriguingly, embryo polarity depends on zygotic gene activities and Wnt signaling. Hence, short-germ insect axis formation is more similar to vertebrates than the mechanism employed by Drosophila.
The distinction of anterior versus posterior is a crucial first step in animal embryogenesis. In the fly Drosophila, this axis is established by morphogenetic gradients contributed by the mother that regulate zygotic target genes. This principle has been considered to hold true for insects in general but is fundamentally different from vertebrates, where zygotic genes and Wnt signaling are required. We investigated symmetry breaking in the beetle Tribolium castaneum, which among insects represents the more ancestral short-germ embryogenesis. We found that maternal Tc-germ cell-less is required for anterior localization of maternal Tc-axin, which represses Wnt signaling and promotes expression of anterior zygotic genes. Both RNAi targeting Tc-germ cell-less or double RNAi knocking down the zygotic genes Tc-homeobrain and Tc-zen1 led to the formation of a second growth zone at the anterior, which resulted in double-abdomen phenotypes. Conversely, interfering with two posterior factors, Tc-caudal and Wnt, caused double-anterior phenotypes. These findings reveal that maternal and zygotic mechanisms, including Wnt signaling, are required for establishing embryo polarity and induce the segmentation clock in a short-germ insect.