Peri-gastrulation development of the dasyurid marsupial Sminthopsis macroura (stripe-faced dunnart) in vitro and evidence for patterning of the epiblast prior to gastrulation

Marsupials are potentially excellent models for the study of gastrulation because of their superficial embryonic area (EA), post-gastrulation implantation and their potential to provide information about the evolution of gastrulation. Very few studies have examined this developmental period in marsupials. Using an established developmental timetable, peri-gastrula stage Sminthopsis macroura blastocysts were collected and described in detail by observations on live blastocysts and by the use of histological and immunohistochemical techniques on fixed blastocysts. Gastrulation in S. macroura shares several aspects common to that of both eutherian mammals and birds, but in terms of tissue arrangement and conceptus form, is more similar to the chick than other mammals. Two methods of culturing EA explants flat without their shell were devised. The techniques will markedly increase the number of possible experimental manipulations, which previously were limited by the presence of blastocyst investments. Exposure of fractions of explants of round, morphologically uniform pre-gastrula stage EA to growth factors or signaling molecules implicated in vertebrate gastrulation suggests that like the chick and mouse, the marsupial epiblast is patterned prior to gastrulation. Of all factors tested, basic fibroblast growth factor (bFGF) had the most prominent effect, promoting cell differentiation, and possible mesoderm formation. Data from explant culture suggests that similar to the chick and mouse, limited specification precedes the onset of gastrulation.