An interplay of transcription factors interprets signalling pathways to define anteroposterior positions along the vertebrate axis. In the hindbrain, these transcription factors prompt the position-appropriate appearance of seven to eight segmental structures, known as rhombomeres (r1-r8). The evolutionarily conserved Cdx caudal-type homeodomain transcription factors help specify the vertebrate trunk and tail but have not been shown to directly regulate hindbrain patterning genes. Mafb (Kreisler, Krml1, valentino), a basic domain leucine zipper transcription factor, is required for development of r5 and r6 and is the first gene to show restricted expression within these two segments. The homeodomain protein vHnf1 (Hnf1b) directly activates Mafb expression. vHnf1 and Mafb share an anterior expression limit at the r4/r5 boundary but vHnf1 expression extends beyond the posterior limit of Mafb and, therefore, cannot establish the posterior Mafb expression boundary. Upon identifying regulatory sequences responsible for posterior Mafb repression, we have used in situ hybridization, immunofluorescence and chromatin immunoprecipitation (ChIP) analyses to determine that Cdx1 directly inhibits early Mafb expression in the neural tube posterior of the r6/r7 boundary, which is the anteriormost boundary of Cdx1 expression in the hindbrain. Cdx1 dependent repression of Mafb is transient. After the 10-somite stage, another mechanism acts to restrict Mafb expression in its normal r5 and r6 domain, even in the absence of Cdx1. Our findings identify Mafb as one of the earliest direct targets of Cdx1 and show that Cdx1 plays a direct role in early hindbrain patterning. Thus, just as Cdx2 and Cdx4 govern the trunk-to-tail transition, Cdx1 may regulate the hindbrain-to-spinal cord transition.

Seasonal changes in the composition and food habits of fish communities in a Sargassum confusum-dominated bed off the coast of Ishikari, Hokkaido, Japan were examined. Twenty-seven species, a total of 997 individuals, were collected by beach seine surveys from June 2002 to August 2003. Numbers of individuals and species increased in spring to fall compared to winter. A cluster analysis based on stomach contents composition showed that the fish communities comprised eight feeding groups: A) phytal amphipods, B) phytal and benthic amphipods, C) benthic polychaetes and phytal amphipods, D) phytal amphipods and isopods, E) benthic copepods and amphipods, F) planktonic copepods, G) phytal mysids and planktonic copepods, and H) phytal mysids feeders. Ontogenetic diet shifts were recognized in Pholidapus dybowskii, Aulichthys japonicus, Hexagrammos otakii and Syngnathus schlegeli, although Blepsias cirrhosus, Neozoarces steindachneri and Zoarchias veneficus were included in nearly the same feeding groups throughout the year. Sebastes schlegelii and Sebastes taczanowskii seemed to change their food habits with the amount of favorite diet. It is suggested that dietary diversity and stable production in the Sargassum bed contribute to the formation of the feeding ground.

Pancreatic endocrine cell differentiation depends on transcription factors that also contribute in adult insulin and glucagon gene expression. Islet cell development was examined in mice lacking MafB, a transcription factor expressed in immature α (glucagon + ) and β (insulin + ) cells and capable of activating insulin and glucagon expression in vitro . We observed that MafB −/− embryos had reduced numbers of insulin + and glucagon + cells throughout development, whereas the total number of endocrine cells was unchanged. Moreover, production of insulin + cells was delayed until embryonic day (E) 13.5 in mutant mice and coincided with the onset of MafA expression, a MafB-related activator of insulin transcription. MafA expression was only detected in the insulin + cell population in MafB mutants, whereas many important regulatory proteins continued to be expressed in insulin − β cells. However, Pdx1, Nkx6.1, and GLUT2 were selectively lost in these insulin-deficient cells between E15.5 and E18.5. MafB appears to directly regulate transcription of these genes, because binding was observed within endogenous control region sequences. These results demonstrate that MafB plays a previously uncharacterized role by regulating transcription of key factors during development that are required for the production of mature α and β cells.

The early transcriptional hierarchy that subdivides the vertebrate hindbrain into seven to eight segments, the rhombomeres (r1–r8), is largely unknown. The Kreisler (MafB, Krml1, Val) gene is earliest gene expressed in an r5/r6-restricted manner and is essential for r5 and r6 development. We have identified the S5 regulatory element that directs early Kreisler expression in the future r5/r6 domain in 0–10 somite stage embryos. variant Hepatocyte Nuclear Factor 1 (vHNF1/HNF1β/LF-3B) is transiently expressed in the r5/r6 domain of 0–10 somite stage embryos and a vHNF1binding site within this element is essential but not sufficient for r5/r6-specific expression. Thus, early inductive events that initiate Kreisler expression are clearly distinct from later-acting ones that modulate its expression levels. This site and some of the surrounding sequences are evolutionarily conserved in the genomic DNA upstream of the Kreisler gene among species as divergent as mouse, humans, and chickens. This provides the first evidence of a direct requirement for vHNF1 in initiation of Kreisler expression, suggests that the role of vHNF1 is evolutionarily conserved, and indicates that vHNF1 collaborates with other transcription factors, which independently bind to the S5 regulatory region, to establish the r5/r6 domain.

Mammalian Polycomb group (PcG) proteins are known to function during the maintenance of spatially restricted expression of Hox cluster genes and cellular proliferation. To understand the molecular basis of PcG functions, it is important to identify the components of mammalian PcG complexes. We isolated mouse YAF2 as a protein that interacts with Ring1B, a known constituent of mammalian PcG complexes. We show that the murine YAF2 locus generates two different transcripts, mYAF2-a and mYAF2-b by alternative splicing of the third exons which encode two YAF2 isoforms of 179 and conceptual 60 amino acids, respectively. At least five exons encoding mYAF2 transcripts are mapped on chromosome 15E3 region. Expression of mYAF2 mRNA was observed in both pre- and postimplantation embryos. In mid-gestation embryos, mYAF2 expression is strongly seen in the region close to the surface ectoderm. Finally, biochemical evidence and colocalization studies in tissue culture cells suggest that the product of the mYAF2 gene is involved in PcG complexes together with Ring1B and/or Ring1A.

NiB and NiWB films fabricated by electroless deposition were evaluated aiming for the application to a metal cap in the copper interconnects technology. The content of B and W was varied by adjusting the concentration of components in electroless deposition baths in order to clarify the effect of co-deposited element on thermal stability of the films. The thermal stability was evaluated by Auger electron spectroscope, X-ray diffractometer (XRD) and sheet resistance measurement. By measuring the variation in sheet resistance with annealing temperature, it was confirmed that the NiB films showed good thermal stability up to 450 °C, whereas the NiWB films deteriorated at 300 °C. The effect of co-deposited element was discussed based on the results obtained by XRD as well as that of sheet resistance measurement.

The products of the Polycomb group of genes form complexes that maintain the state of transcriptional repression of several genes with relevance to development and in cell proliferation. We have identified Ring1B, the product of the Ring1B gene (Rnf2 - Mouse Genome Informatics), by means of its interaction with the Polycomb group protein Mel18. We describe biochemical and genetic studies directed to understand the biological role of Ring1B. Immunoprecipitation studies indicate that Ring1B form part of protein complexes containing the products of other Polycomb group genes, such as Rae28/Mph1 and M33, and that this complexes associate to chromosomal DNA. We have generated a mouse line bearing a hypomorphic Ring1B allele, which shows posterior homeotic transformations of the axial skeleton and a mild derepression of some Hox genes (Hoxb4, Hoxb6 and Hoxb8) in cells anterior to their normal boundaries of expression in the mesodermal compartment. By contrast, the overexpression of Ring1B in chick embryos results in the repression of Hoxb9 expression in the neural tube. These results, together with the genetic interactions observed in compound Ring1B/Mel18 mutant mice, are consistent with a role for Ring1B in the regulation of Hox gene expression by Polycomb group complexes.

The precise expression patterns of two IgLON genes, CEPU‐1 and limbic system‐associated membrane protein (LAMP), were studied during early embryogenesis. It was found that expression of both was localized to restricted regions of the brain and neural crest. In the developing neural tube, CEPU‐1 was expressed in the isthmus and a restricted region of the hindbrain, whereas LAMP was expressed in the anterior midbrain. Most neural crest cells expressed LAMP, whereas CEPU‐1 expression was limited to crest cells derived from the hindbrain. These results suggest that members of the IgLON family have important roles during embryogenesis, particularly in brain formation and differentiation.