Neurodevelopment Genes in Lampreys Reveal Trends for Forebrain Evolution in Craniates

The forebrain is the brain region which has undergone the most dramatic changes through vertebrate evolution. Analyses conducted in lampreys are essential to gain insight into the broad ancestral characteristics of the forebrain at the dawn of vertebrates, and to understand the molecular basis for the diversifications that have taken place in cyclostomes and gnathostomes following their splitting. Here, we report the embryonic expression patterns of 43 lamprey genes, coding for transcription factors or signaling molecules known to be involved in cell proliferation, stemcellness, neurogenesis, patterning and regionalization in the developing forebrain. Systematic expression patterns comparisons with model organisms highlight conservations likely to reflect shared features present in the vertebrate ancestors. They also point to changes in signaling systems –pathways which control the growth and patterning of the neuroepithelium-, which may have been crucial in the evolution of forebrain anatomy at the origin of vertebrates

How blind are they? Phototactic responses in stygobiont diving beetles (Coleoptera: Dytiscidae) from calcrete aquifers of Western Australia

Subterranean water beetles endemic to groundwater calcretes of Western Australia exhibit convergent traits typical of troglomorphic arthropods, including loss of eyes, pigmentation and wings. As these dytiscid species are estimated to have been isolated underground in permanent darkness for over three million years, it is predicted that they will completely lack phototactic responses. We tested this hypothesis by analysing the behaviour of six subterranean beetle species within an observational arena with dark and light hemispheres. Scan samples at 1 min intervals and total time spent on each hemisphere were recorded over a 20 min period, testing at least 15 individuals per species. We quantified behaviour as an index (dark ratio) so that individual species in this, and future, studies can be consistently compared. Results analysed as both categorical and absolute proportion of time spent in each hemisphere suggest negative phototaxis in Paroster macrosturtensis. The remaining five species did not display any preference for either light or dark hemispheres. These results raise the possibility that some ancestral Paroster species may have exhibited negative phototactic behaviour prior to subterranean colonization. The retention of such a behavioural trait in lightless environments could represent the maintenance for some unknown pleiotropic function. Alternatively, it is possible that insufficient time has passed for neutral processes to render photoreception genes and phototactic behaviours non-functional. Our study adds to a growing body of evidence that implies highly troglomorphic animals may have evolved from ancestral species that exhibited negative phototaxis as a preadaptation to living in permanent darkness

Two lamprey Hedgehog genes share non-coding regulatory sequences and expression patterns with gnathostome Hedgehogs.

Hedgehog (Hh) genes play major roles in animal development and studies of their evolution, expression and function point to major differences among chordates. Here we focused on Hh genes in lampreys in order to characterize the evolution of Hh signalling at the emergence of vertebrates. Screening of a cosmid library of the river lamprey Lampetra fluviatilis and searching the preliminary genome assembly of the sea lamprey Petromyzon marinus indicate that lampreys have two Hh genes, named Hha and Hhb. Phylogenetic analyses suggest that Hha and Hhb are lamprey-specific paralogs closely related to Sonic/Indian Hh genes. Expression analysis indicates that Hha and Hhb are expressed in a Sonic Hh-like pattern. The two transcripts are expressed in largely overlapping but not identical domains in the lamprey embryonic brain, including a newly-described expression domain in the nasohypophyseal placode. Global alignments of genomic sequences and local alignment with known gnathostome regulatory motifs show that lamprey Hhs share conserved non-coding elements (CNE) with gnathostome Hhs albeit with sequences that have significantly diverged and dispersed. Functional assays using zebrafish embryos demonstrate gnathostome-like midline enhancer activity for CNEs contained in intron2. We conclude that lamprey Hh genes are gnathostome Shh-like in terms of expression and regulation. In addition, they show some lamprey-specific features, including duplication and structural (but not functional) changes in the intronic/regulatory sequences

Phylogenomic analysis and expression patterns of large Maf genes in Xenopus tropicalis provide new insights into the functional evolution of the gene family in osteichthyans.

We have performed an exhaustive characterization of the large Maf family of basic leucine zipper transcription factors in vertebrates using the genome data available, and studied the embryonic expression patterns of the four paralogous genes thus identified in Xenopus tropicalis. Our phylogenetic analysis shows that, in osteichthyans, the large Maf family contains four orthology classes, MafA, MafB, c-Maf and Nrl, which have emerged in vertebrates prior to the split between actinopterygians and sarcopterygians. It leads to the unambiguous assignment of the Xenopus laevis XLmaf gene, previously considered a MafA orthologue, to the Nrl class, the identification of the amphibian MafA and c-Maf orthologues and the identification of the zebrafish Nrl gene. The four X. tropicalis paralogues display partially redundant but nevertheless distinct expression patterns in the somites, developing hindbrain, pronephros, ventral blood island and lens. Comparisons with the data available in the mouse, chick and zebrafish show that these large Maf expression territories are highly conserved among osteichthyans but also highlight a number of differences in the timing of large Maf gene expression, the precise extent of some labelled territories and the combinations of paralogues transcribed in some organs. In particular, the availability of robust phylogenies leads to a reinterpretation of previous expression pattern comparisons, suggesting an important part for function shuffling within the gene family in the developing lens. These data highlight the importance of exhaustive characterizations of gene families for comparative analyses of the genetic mechanisms, which control developmental processes in vertebrates

A question of homology for chordate adhesive organs

The larvae of aquatic vertebrates sometimes possess a transient, mucus-secreting gland on their heads. The most studied of these organs is the Xenopus cement gland. The tadpoles use it to attach to plants or to the water surface, supposedly to hide from predators and save energy before they can swim or feed. Moreover their gland, being innervated by trigeminal fibres, also mediates a locomotor stopping response when the larvae encounter an obstacle. We have described an equivalent organ on the head of the teleost Astyanax mexicanus, that we have called the casquette because of its shape and position on the larval head. The casquette is transient, sticky, secretes mucus, is innervated by the trigeminal ganglion, has an inhibitory function on larval swimming behavior, and expresses Bmp4 and Pitx1/2 during embryogenesis. Here we further discuss the nature of the equivalence between the frog cement gland and the fish casquette, and highlight the usefulness of non-conventional model species to decipher developmental and evolutionary mechanisms of morphological variations

A supernumerary "B-sex" chromosome drives male sex determination in the Pachón cavefish, Astyanax mexicanus.

Sex chromosomes are generally derived from a pair of classical type-A chromosomes, and relatively few alternative models have been proposed up to now. <sup>1</sup> <sup>,</sup> <sup>2</sup> B chromosomes (Bs) are supernumerary and dispensable chromosomes with non-Mendelian inheritance found in many plant and animal species <sup>3</sup> <sup>,</sup> <sup>4</sup> that have often been considered as selfish genetic elements that behave as genome parasites. <sup>5</sup> <sup>,</sup> <sup>6</sup> The observation that in some species Bs can be either restricted or predominant in one sex <sup>7-14</sup> raised the interesting hypothesis that Bs could play a role in sex determination. <sup>15</sup> The characterization of putative B master sex-determining (MSD) genes, however, has not yet been provided to support this hypothesis. Here, in Astyanax mexicanus cavefish originating from Pachón cave, we show that Bs are strongly male predominant. Based on a high-quality genome assembly of a B-carrying male, we characterized the Pachón cavefish B sequence and found that it contains two duplicated loci of the putative MSD gene growth differentiation factor 6b (gdf6b). Supporting its role as an MSD gene, we found that the Pachón cavefish gdf6b gene is expressed specifically in differentiating male gonads, and that its knockout induces male-to-female sex reversal in B-carrying males. This demonstrates that gdf6b is necessary for triggering male sex determination in Pachón cavefish. Altogether these results bring multiple and independent lines of evidence supporting the conclusion that the Pachón cavefish B is a "B-sex" chromosome that contains duplicated copies of the gdf6b gene, which can promote male sex determination in this species