Exogenous retinoic acid induces digit reduction in opossums (Monodelphis domestica) by disrupting cell death and proliferation, and apical ectodermal ridge and zone of polarizing activity function.

BACKGROUND: Retinoic acid (RA) is a vitamin A derivative. Exposure to exogenous RA generates congenital limb malformations (CLMs) in species from frogs to humans. These CLMs include but are not limited to oligodactyly and long-bone hypoplasia. The processes by which exogenous RA induces CLMs in mammals have been best studied in mouse, but as of yet remain unresolved. METHODS: We investigated the impact of exogenous RA on the cellular and molecular development of the limbs of a nonrodent model mammal, the opossum Monodelphis domestica. Opossums exposed to exogenous retinoic acid display CLMs including oligodactly, and results are consistent with opossum development being more susceptible to RA-induced disruptions than mouse development. RESULTS: Exposure of developing opossums to exogenous RA leads to an increase in cell death in the limb mesenchyme that is most pronounced in the zone of polarizing activity, and a reduction in cell proliferation throughout the limb mesenchyme. Exogenous RA also disrupts the expression of Shh in the zone of polarizing activity, and Fgf8 in the apical ectodermal ridge, and other genes with roles in the regulation of limb development and cell death. CONCLUSION: Results are consistent with RA inducing CLMs in opossum limbs by disrupting the functions of the apical ectodermal ridge and zone of polarizing activity, and driving an increase in cell death and reduction of cell proliferation in the mesenchyme of the developing limb.

Development of the marsupial shoulder girdle complex: a case study in Monodelphis domestica.

During their embryogenesis, marsupials transiently develop a unique structure, the shoulder arch, which provides the structural support and muscle-attachments necessary for the newborn's crawl to the teat. One of the most pronounced and functionally important aspects of the shoulder arch is an enlarged coracoid. The goal of this study is to determine the molecular basis of shoulder arch formation in marsupials. To achieve this goal, this study investigates the relative expression of several genes with known roles in shoulder girdle morphogenesis in a marsupial-the opossum, Monodelphis domestica-and a placental, the mouse, Mus musculus. Results indicate that Hoxc6, a gene involved in coracoid patterning, is expressed for a longer period of time and at higher levels in opossum relative to mouse. Functional manipulation suggests that these differences in Hoxc6 expression are independent of documented differences in retinoic acid signaling in opossum and mouse forelimbs. Results also indicate that Emx2, a gene involved in scapular blade condensation, is upregulated in opossum relative to mouse. However, several other genes involved in shoulder girdle patterning (e.g., Gli3, Pax1, Pbx1, Tbx15) are comparably expressed in these species. These findings suggest that the upregulation of Hoxc6 and Emx2 occurs through independent genetic modifications in opossum relative to mouse. In summary, this study documents a correlation between gene expression and the divergent shoulder girdle morphogenesis of marsupial (i.e., opossum) and placental (i.e., mouse) mammals, and thereby provides a foundation for future research into the genetic basis of shoulder girdle morphogenesis in marsupials. Furthermore, this study supports the hypothesis that the mammalian shoulder girdle is a highly modular structure whose elements are relatively free to evolve independently.