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Generation of Dispersed Presomitic Mesoderm Cell Cultures for Imaging of the Zebrafish Segmentation Clock in Single Cells
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Anteroposterior and dorsoventral patterning are coordinated by an identical patterning clock.

Megumi Hashiguchi1, Mary C Mullins

  • 1Department of Cell and Developmental Biology, University of Pennsylvania, Perelman School of Medicine, 421 Curie Blvd., Philadelphia, PA 19104-6058, USA.

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|March 29, 2013
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Summary

Vertebrate body plan formation relies on coordinated tissue patterning. This study reveals that dorsoventral and anteroposterior patterning in zebrafish share a common molecular clock, ensuring simultaneous positional and temporal information acquisition.

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Area of Science:

  • Developmental Biology
  • Molecular Biology
  • Genetics

Background:

  • Vertebrate body plan establishment requires precise temporal coordination of tissue patterning along the anterior-posterior (AP) and dorsoventral (DV) axes.
  • Previous research indicated DV patterning progresses anteriorly to posteriorly via Bone Morphogenetic Protein (BMP) signaling.
  • The integration of AP and DV patterning mechanisms remained incompletely understood.

Purpose of the Study:

  • To investigate the temporal coordination between DV and AP patterning along the zebrafish body axes.
  • To elucidate the molecular mechanisms integrating BMP signaling with other key developmental pathways (FGF, Wnt, retinoic acid) in patterning.
  • To propose a model for the simultaneous temporal coordination of DV and AP patterning.

Main Methods:

  • Manipulation of FGF, Wnt, and retinoic acid signaling pathways, combined with BMP signaling inhibition, at various developmental time points in zebrafish embryos.
  • Analysis of phosphorylated Smad1/5 (P-Smad1/5) localization and function in response to signaling pathway alterations.
  • Site-directed mutagenesis of Smad5 linker phosphorylation sites to assess functional consequences on DV patterning.

Main Results:

  • Zebrafish DV patterning along the AP axis is temporally coordinated with AP patterning by an identical patterning clock.
  • FGF/MAPK signaling, not Wnt/GSK3, phosphorylates the Smad1/5 linker region in a ventral vegetal gastrula region, potentially coordinating DV and AP patterning without P-Smad1/5 degradation.
  • Altering MAPK phosphorylation sites on Smad5 resulted in precocious DV tissue patterning along the AP axis during gastrulation.

Conclusions:

  • Dorsoventral and anteroposterior patterning in zebrafish are intimately coordinated by a shared molecular clock.
  • FGF/MAPK signaling plays a crucial role in integrating DV and AP patterning through Smad1/5 linker phosphorylation.
  • This coordination allows cells to acquire both positional and temporal information simultaneously during vertebrate development.