During vertebrate development the dorsal gastrula or Spemann-Mangold organizer orchestrates axis formation largely by limiting the ventralizing and posteriorizing activity of bone morphogenetic proteins (BMPs). In mouse and Xenopus laevis , genes encoding the zinc finger transcriptional repressor Prdm1/Blimp1 (PR domain containing 1, with ZNF domain; previously named B lymphocyte-induced maturation protein 1) were recently shown to be expressed in the visceral endoderm and anterior endomesoderm, respectively,and the prechordal plate of gastrula stage embryos. Later in development Prdm1/Blimp1 is expressed in many other tissues, including pharyngeal arches, limb buds, otic vesicles, photoreceptor cell layer, slow muscle and cloaca. Based on misexpression and dominant-negative studies, Prdm1/Blimp1 was proposed to promote anterior endomesoderm and head development in Xenopus laevis . Here we report the isolation and functional characterization of zebrafish prdm1 exhibiting a dynamic and evolutionarily conserved expression pattern. Misexpression of prdm1 inhibits the formation of dorsoanterior structures and reduces expression of chordin , which encodes a BMP antagonist. Conversely, interference with Prdm1 translation using antisense morpholino oligonucleotides, increases chordin expression, while reducing expression of Bmp genes, and consequently dorsalizing the embryo. At the end of the gastrula period, prdm1 morphant embryos have enlarged animal-vegetal and anteroposterior embryonic axes. This altered embryo morphology is associated with augmented extension movements of dorsal tissues and normal posterior migration of ventral tissues. Additionally, Prdm1 activity is essential for proper development of slow muscle, the photoreceptor cell layer, branchial arches and pectoral fins. Our studies reveal essential roles for prdm1 in limiting the function of the gastrula organizer and regulating cell fate specification and morphogenetic processes in precise correspondence with its intricate expression pattern.

Little is known about the factors that control the specification of the mid-hindbrain domain (MHD) within the vertebrate embryonic neural plate. Because the head-trunk junction of the Drosophila embryo and the MHD have patterning similarities, we have searched for vertebrate genes related to the Drosophila head gap gene buttonhead ( btd ), which in the fly specifies the head-trunk junction. We report here the identification of a zebrafish gene which, like btd , encodes a zinc-finger transcriptional activator of the Sp-1 family (hence its name, bts1 for btd/Sp-related-1 ) and shows a restricted expression in the head. During zebrafish gastrulation, bts1 is transcribed in the posterior epiblast including the presumptive MHD, and precedes in this area the expression of other MHD markers such as her5 , pax2.1 and wnt1 . Ectopic expression of bts1 combined to knock-down experiments demonstrate that Bts1 is both necessary and sufficient for the induction of pax2.1 within the anterior neural plate, but is not involved in regulating her5 , wnt1 or fgf8 expression. Our results confirm that early MHD development involves several genetic cascades that independently lead to the induction of MHD markers, and identify Bts1 as a crucial upstream component of the pathway selectively leading to pax2.1 induction. In addition, they imply that flies and vertebrates, to control the development of a boundary embryonic region, have probably co-opted a similar strategy: the restriction to this territory of the expression of a Btd/Sp-like factor.

The dorsal ectoderm of the vertebrate gastrula was proposed by Nieuwkoop to be specified towards an anterior neural fate by an activation signal, with its subsequent regionalization along the anteroposterior (AP) axis regulated by a graded transforming activity, leading to a properly patterned forebrain, midbrain, hindbrain and spinal cord. The activation phase involves inhibition of BMP signals by dorsal antagonists, but the later caudalization process is much more poorly characterized. Explant and overexpression studies in chick, Xenopus , mouse and zebrafish implicate lateral/paraxial mesoderm in supplying the transforming influence, which is largely speculated to be a Wnt family member. We have analyzed the requirement for the specific ventrolaterally expressed Wnt8 ligand in the posteriorization of neural tissue in zebrafish wild-type and Nodal-deficient embryos (Antivin overexpressing or cyclops;squint double mutants), which show extensive AP brain patterning in the absence of dorsal mesoderm. In different genetic situations that vary the extent of mesodermal precursor formation, the presence of lateral wnt8 -expressing cells correlates with the establishment of AP brain pattern. Cell tracing experiments show that the neuroectoderm of Nodal-deficient embryos undergoes a rapid anterior-to-posterior transformation in vivo during a short period at the end of the gastrula stage. Moreover, in both wild-type and Nodal-deficient embryos, inactivation of Wnt8 function by morpholino (MO wnt8 ) translational interference dose-dependently abrogates formation of spinal cord and posterior brain fates, without blocking ventrolateral mesoderm formation. MO wnt8 also suppresses the forebrain deficiency in bozozok mutants, in which inactivation of a homeobox gene causes ectopic wnt8 expression. In addition, the bozozok forebrain reduction is suppressed in bozozok;squint;cyclops triple mutants, and is associated with reduced wnt8 expression, as seen in cyclops;squint mutants. Hence, whereas boz and Nodal signaling largely cooperate in gastrula organizer formation, they have opposing roles in regulating wnt8 expression and forebrain specification. Our findings provide strong support for a model of neural transformation in which a planar gastrula-stage Wnt8 signal, promoted by Nodal signaling and dorsally limited by Bozozok, acts on anterior neuroectoderm from the lateral mesoderm to produce the AP regional patterning of the CNS.