The role of planar and early vertical signaling in patterning the expression of Hoxb-1 in Xenopus.
In this paper we examine the contributions of planar and vertical signaling to the patterning of gene expression in neural development and we examine the routes of this neural induction. We have examined how the expression of Xenopus homeobox gene, Hoxb-1, is regulated by instruction from the mesoderm and/or endoderm and ask whether this instruction is by the vertical or planar routes. We investigated normal expression patterns of Hoxb-1 during early Xenopus development and Hoxb-1 expression in sandwich explants of the dorsal marginal zone, which putatively allow only planar signals to pass from the mesodermal and endodermal tissue (Spemann's organizer) to the prospective neural tissue. In the latter case we found significant variability of expression. Observations during dissections suggested that variable degrees of invasion of the mesodermal-endodermal tissue at the leading edge of the mesodermal mantle might be the cause of this variability. Alternatively, differing lengths of time that the prospective neural region spends in planar contact with tissues of the lateral or ventral regions of the embryo could also contribute to this variability. Analysis of staged Keller sandwich explants, "skewered" sandwiches, in which the degree of contact with underlying, involuted mesoderm-endodermal tissues was marked, and "over-the-pole" and "giant" sandwich explants, in which the degree of planar contact with lateral or ventral tissues was normalized, suggests that both planar and vertical signals are involved in induction and patterning of Hoxb-1 expression. The shift in Hoxb-1 expression from a broad, diffuse pattern to a local, focused pattern, characteristic of the ultimate expression pattern in vivo, does not reflect variable degrees of contact with ventral or lateral tissues, but rather reflects early vertical contact with underlying mesodermal-endodermal tissues. We observed such contact at early gastrula stages (stages 10 to 10+), stages commonly assumed not to have the potential for vertical signaling. As the bottle cells first begin to form, at stage 10-, a massive rotation of the lower involuting marginal zone occurs around an internal lip ("levre interne," Nieuwkoop and Florschutz, 1950). This rotation initiates the formation of the Cleft of Brachet from the floor of the blastocoele and brings the prospective mesoderm and endoderm at the leading edge of the marginal zone into vertical apposition with the prospective neural region quite early in gastrulation. The consequence and importance of recognizing these early internal rearrangements are that it pushes backward the time at which potential vertical inductive interactions between mesoderm and neurectoderm can occur. This means that a purely planar inductive situation can cease to exist as early as the inception of bottle cell formation and that neural patterning through vertical induction starts at the very beginning of gastrulation.
Poznanski
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Molecular and Cell Biology, University of California, Berkeley 94720, USA.
Keller
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R
D000595:Amino Acid Sequence; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002478:Cells, Cultured; D017124:Conserved Sequence / Q000235:genetics; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000378:metabolism; D051767:Early Growth Response Protein 2; D004625:Embryo, Nonmammalian; D018507:Gene Expression Regulation, Developmental; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000378:metabolism; D009420:Nervous System / Q000196:embryology* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction / Q000502:physiology*; D014157:Transcription Factors / Q000378:metabolism; D014981:Xenopus
D015415:Biomarkers; D017931:DNA Primers; D004268:DNA-Binding Proteins; D051767:Early Growth Response Protein 2; C101121:HOXB1 homeodomain protein; D018398:Homeodomain Proteins; D009419:Nerve Tissue Proteins; D014157:Transcription Factors