Fgf10+ progenitors give rise to the chick hypothalamus by rostral and caudal growth and differentiation.

Classical descriptions of the hypothalamus divide it into three rostro-caudal domains but little is known about their embryonic origins. To investigate this, we performed targeted fate-mapping, molecular characterisation and cell cycle analyses in the embryonic chick. Presumptive hypothalamic cells derive from the rostral diencephalic ventral midline, lie above the prechordal mesendoderm and express Fgf10Fgf10+ progenitors undergo anisotropic growth: those displaced rostrally differentiate into anterior cells, then those displaced caudally differentiate into mammillary cells. A stable population of Fgf10+ progenitors is retained within the tuberal domain; a subset of these gives rise to the tuberal infundibulum - the precursor of the posterior pituitary. Pharmacological approaches reveal that Shh signalling promotes the growth and differentiation of anterior progenitors, and also orchestrates the development of the infundibulum and Rathke's pouch - the precursor of the anterior pituitary. Together, our studies identify a hypothalamic progenitor population defined by Fgf10 and highlight a role for Shh signalling in the integrated development of the hypothalamus and pituitary.

Pax3 synergizes with Gli2 and Zic1 in transactivating the Myf5 epaxial somite enhancer.

Both Glis, the downstream effectors of hedgehog signaling, and Zic transcription factors are required for Myf5 expression in the epaxial somite. Here we demonstrate a novel synergistic interaction between members of both families and Pax3, a paired-domain transcription factor that is essential for both myogenesis and neural crest development. We show that Pax3 synergizes with both Gli2 and Zic1 in transactivating the Myf5 epaxial somite (ES) enhancer in concert with the Myf5 promoter. This synergy is dependent on conserved functional domains of the proteins, as well as on a novel homeodomain motif in the Myf5 promoter and the essential Gli motif in the ES enhancer. Importantly, overexpression of Zic1 and Pax3 in the 10T1/2 mesodermal cell model results in enrichment of these factors at the endogenous Myf5 locus and induction of Myf5 expression. In our previous work, we showed that by enhancing nuclear translocation of Gli factors, Zics provide spatiotemporal patterning for Gli family members in the epaxial induction of Myf5 expression. Our current study indicates a complementary mechanism in which association with DNA-bound Pax3 strengthens the ability of both Zic1 and Gli2 to transactivate Myf5 in the epaxial somite.

Six3 inactivation causes progressive caudalization and aberrant patterning of the mammalian diencephalon.

The homeobox gene Six3 represses Wnt1 transcription. It is also required in the anterior neural plate for the development of the mammalian rostral forebrain. We have now determined that at the 15- to 17-somite stage, the prospective diencephalon is the most-anterior structure in the Six3-null brain, and Wnt1 expression is anteriorly expanded. Consequently, the brain caudalizes, and at the 22- to 24-somite stage, the prospective thalamic territory is the most-anterior structure. At around E11.0, the pretectum replaces this structure. Analysis of Six3;Wnt1 double-null mice revealed that Six3-mediated repression of Wnt1 is necessary for the formation of the rostral diencephalon and that Six3 activity is required for the formation of the telencephalon. These results provide insight into the mechanisms that establish anteroposterior identity in the developing mammalian brain.

Genetic analysis of somite formation in laboratory fish models.

The repeated appearance of somites is one of the most fascinating aspects of vertebrate embryogenesis. Recent studies identified complex regulatory circuits that provide the molecular basis for the "clock and wave front" model, postulated almost 30 years ago by Cooke and Zeeman. The highly coordinated process of somite formation involves several networks of molecular cascades including the Delta/Notch, Wnt, FGF and retinoid signalling pathways. Studies in mouse, Xenopus and especially chicken over the last decade have helped to understand the role and interactions of these pathways in somitogenesis. More recently, this has been supplemented by experiments in zebrafish. This animal model offers the possibility of performing large scale mutagenesis screens to identify novel factors and pathways involved in somitogenesis. Molecular cloning of zebrafish somite mutants mainly resulted in genes that belong to the Delta/Notch pathway and therefore underlined the importance of this pathway during somitogenesis. The fact that other pathways have not yet been identified by genetic screening in this species was assumed to be caused by functional redundancy of duplicated genes in zebrafish. In 2000, a large-scale mutagenesis screen has been initiated in Kyoto, Japan using the related teleost medaka (Oryzias latipes). In this screen, mutants with unique phenotypes have been identified, which have not been described in zebrafish or mouse. In this chapter, we will review the progress that has been made in understanding the molecular control of somite formation in zebrafish and will discuss recent efforts to screen for novel phenotypes using medaka somitogenesis mutants.