The notochord curvature in medaka (Oryzias latipes) embryos as a response to ultraviolet A irradiation.

In the present work, the destructive effects of ultraviolet A (UVA; 366nm) irradiation on the developmental stages of Japanese medaka (Oryzias latipes) are revealed in terms of hatching success, mortality rate, and morphological malformations (yolk sac edema, body curvature, fin blistering, and dwarfism). Fertilized eggs in stage 4 were exposed to 15, 30, and 60min/day UVA for 3days in replicates. Fish were staged and aged following the stages established by Iwamatsu [1]. We observed and recorded the hatching time and deformed and dead embryos continuously. The hatching time was prolonged and the deformed and dead embryos numbers were increased by UVA dose increase. At stage 40, samples from each group were fixed to investigate their morphology and histopathology. Some morphological malformations were recorded after UVA exposure in both strains. Histopathological changes were represented as different shapes of curvature in notochord with collapse. The degree of collapsation was depended on the dose and time of UVA exposure. Our findings show that exposure to UVA irradiation caused less vertebral column curvature in medaka fry. Moreover, p53-deficient embryos were more tolerant than those of wild-type (Hd-rR) Japanese medaka. This study indicated the dangerous effects of the UVA on medaka.

X-ray micro-diffraction studies on biological samples at the BioCAT Beamline 18-ID at the Advanced Photon Source.

The small source sizes of third-generation synchrotron sources are ideal for the production of microbeams for diffraction studies of crystalline and non-crystalline materials. While several such facilities have been available around the world for some time now, few have been optimized for the handling of delicate soft-tissue specimens under cryogenic conditions. Here the development of a new X-ray micro-diffraction instrument at the Biophysics Collaborative Access Team beamline 18-ID at the Advanced Photon Source, and its use with newly developed cryo-diffraction techniques for soft-tissue studies, are described. The combination of the small beam sizes delivered by this instrument, the high delivered flux and successful cryo-freezing of rat-tail tendon has enabled us to record data to better than 4 Šresolution. The ability to quickly raster scan samples in the beam allows selection of ordered regions in fibrous samples for markedly improved data quality. Examples of results of experiments obtainable using this instrument are presented.

Collagen type XI alpha1 may be involved in the structural plasticity of the vertebral column in Atlantic salmon (Salmo salar L.).

Atlantic salmon (Salmo salar L.) vertebral bone displays plasticity in structure, osteoid secretion and mineralization in response to photoperiod. Other properties of the vertebral bone, such as mineral content and mechanical strength, are also associated with common malformations in farmed Atlantic salmon. The biological mechanisms that underlie these changes in bone physiology are unknown, and in order to elucidate which factors might be involved in this process, microarray assays were performed on vertebral bone of Atlantic salmon reared under natural or continuous light. Eight genes were upregulated in response to continuous light treatment, whereas only one of them was upregulated in a duplicate experiment. The transcriptionally regulated gene was predicted to code for collagen type XI alpha1, a protein known to be involved in controlling the diameter of fibrillar collagens in mammals. Furthermore, the gene was highly expressed in the vertebrae, where spatial expression was found in trabecular and compact bone osteoblasts and in the chordoblasts of the notochordal sheath. When we measured the expression level of the gene in the tissue compartments of the vertebrae, the collagen turned out to be 150 and 25 times more highly expressed in the notochord and compact bone respectively, relative to the expression in the trabecular bone. Gene expression was induced in response to continuous light, and reduced in compressed vertebrae. The downregulation in compressed vertebrae was due to reduced expression in the compact bone, while expression in the trabecular bone and the notochord was unaffected. These data support the hypothesis that this gene codes for a presumptive collagen type XI alpha1, which may be involved in the regulatory pathway leading to structural adaptation of the vertebral architecture.

Synergistic effects of Vg1 and Wnt signals in the specification of dorsal mesoderm and endoderm.

In amphibians, dorsoventral asymmetry is established by cortical rotation, a cytoplasmic rearrangement in the egg which activates a dorsal determinant on one side of the zygote. This determinant has been proposed to be either Vgl, an endodermally derived molecule that can directly induce ectoderm to form dorsal mesoderm, or a member of the Wnt family, which patterns the ectoderm such that it forms dorsal mesoderm in response to ventral inductive signals. In this study, we have investigated whether the endogenous dorsal determinant(s) functions as a direct inducer of dorsal mesoderm (Vg1-like) or whether it acts to pattern the response of ectoderm to inductive signals (Wnt-like). We report here that cortical rotation enhances both the dorsal-inductive activity of endodermal cells and the response of ectodermal cells to endogenous inductive signals and that both of these activities are required for notochord induction in ectoderm/endoderm recombinants. While ectopically expressed Xwnt-8b can substitute for the dorsalizing signals activated in either ectoderm or endoderm, and can allow notochord formation in recombinants, Vg1 alone is not sufficient to induce notochord in ectodermal explants in the absence of signals activated by cortical rotation. Coexpression of Xwnt-8b along with Vg1 restores ectodermal competence to form notochord. Finally, in endodermal explants, ectopically expressed Xwnt-8b, but not Vg1, can divert the fate of ventral endodermal cells along a dorsal pathway. Thus, while Vg1 is most likely required for induction of mesoderm in vivo, our data suggest that a maternal Wnt-like signal acts synergistically with Vg1 to specify a dorsal fate not only in the mesoderm, but also in the endoderm.