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Low proliferative and high migratory activity in the area of Brachyury expressing mesoderm progenitor cells in the gastrulating rabbit embryo

Christoph Viebahn^1,2,, Christof Stortz², Sally A. Mitchell³ and Martin Blum^3,*

¹ Institute of Anatomy and Cell Biology, Martin-Luther-University Halle-Wittenberg, Germany
² Institute of Anatomy, Friedrich-Wilhelms-University Bonn, Germany
³ Institute of Toxicology and Genetics, Forschungszentrum Karlsruhe, Germany
^* Present address: Institute of Zoology, University of Stuttgart-Hohenheim, Germany

Author for correspondence (e-mail: christoph.viebahn{at}medizin.uni-halle.de)

Accepted 19 February 2002

General mechanisms initiating the gastrulation process in early animal development are still elusive, not least because embryonic morphology differs widely among species. The rabbit embryo is revived here as a model to study vertebrate gastrulation, because its relatively simple morphology at the appropriate stages makes interspecific differences and similarities particularly obvious between mammals and birds. Three approaches that centre on mesoderm specification as a key event at the start of gastrulation were chosen.

(1) A cDNA fragment encoding 212 amino acids of the rabbit Brachyury gene was cloned by RT-PCR and used as a molecular marker for mesoderm progenitors. Whole-mount in situ hybridisation revealed single Brachyury-expressing cells in the epiblast at 6.2 days post conception, i.e. several hours before the first ingressing mesoderm cells can be detected histologically. With the anterior marginal crescent as a landmark, these mesoderm progenitors are shown to lie in a posterior quadrant of the embryonic disc, which we call the posterior gastrula extension (PGE), for reasons established during the following functional analysis.

(2) Vital dye (DiI) labelling in vitro suggests that epiblast cells arrive in the PGE from anterior parts of the embryonic disc and then move within this area in a complex pattern of posterior, centripetal and anterior directions to form the primitive streak.

(3) BrdU labelling shows that proliferation is reduced in the PGE, while the remaining anterior part of the embryonic disc contains several areas of increased proliferation. These results reveal similarities with the chick with respect to Brachyury expression and cellular migration. They differ, however, in that local differences in proliferation are not seen in the pre-streak avian embryo. Rather, rabbit epiblast cells start mesoderm differentiation in a way similar to Drosophila, where a transient downregulation of proliferation initiates mesoderm differentiation and, hence, gastrulation.

Key words: Primitive streak, Epiblast, Anteroposterior axis, Gastrula, BrdU, DiI, Lagomorph

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