Clustered brachiopod Hox genes are not expressed collinearly and are associated with lophotrochozoan novelties.

Abstract

Temporal collinearity is often considered the main force preserving Hox gene clusters in animal genomes. Studies that combine genomic and gene expression data are scarce, however, particularly in invertebrates like the Lophotrochozoa. As a result, the temporal collinearity hypothesis is currently built on poorly supported foundations. Here we characterize the complement, cluster, and expression of Hox genes in two brachiopod species, <i>Terebratalia transversa</i> and <i>Novocrania anomala</i><i>T. transversa</i> has a split cluster with 10 genes (<i>lab</i>, <i>pb</i>, <i>Hox3</i>, <i>Dfd</i>, <i>Scr</i>, <i>Lox5</i>, <i>Antp</i>, <i>Lox4</i>, <i>Post2</i>, and <i>Post1</i>), whereas <i>N. anomala</i> has 9 genes (apparently missing <i>Post1</i>). Our in situ hybridization, real-time quantitative PCR, and stage-specific transcriptomic analyses show that brachiopod Hox genes are neither strictly temporally nor spatially collinear; only <i>pb</i> (in <i>T. transversa</i>), <i>Hox3</i> (in both brachiopods), and <i>Dfd</i> (in both brachiopods) show staggered mesodermal expression. Thus, our findings support the idea that temporal collinearity might contribute to keeping Hox genes clustered. Remarkably, expression of the Hox genes in both brachiopod species demonstrates cooption of Hox genes in the chaetae and shell fields, two major lophotrochozoan morphological novelties. The shared and specific expression of Hox genes, together with <i>Arx</i>, <i>Zic</i>, and Notch pathway components in chaetae and shell fields in brachiopods, mollusks, and annelids provide molecular evidence supporting the conservation of the molecular basis for these lophotrochozoan hallmarks.