Molecular Methods in Developmental Biology

This book is primarily aimed at newcomers to the field of developmental biology, but is also useful to experienced workers. In this volume, the South African clawed frog Xenopus Iaevis and the zebrafish Brachydanio rerio serve as the model organisms for the developmental studies. Both species are well suited to investigations of early vertebrate development because of their embryos develop rapidly and externally, but each also has its set of disadvantages. This methods manual includes experimental approaches which can be applied to both species, since many laboratories also use these two models for similar studies. For example, the approaches for the analysis of steady-state mRNA levels by RT-PCR or PNase protection are identical for Xenopus and zebrafish, but for whole-mount in situ hybridization, two tissue explant and transplant techniques, quantitative and spatial analysis of mRNA and immunohistochemistry. The final chapters focus on more specialized methods for expression and promoter analysis, such as the analysis of transcription control in early development. The methods and techniques presented in this volume will provide a sound basis for the majority of studies in this field.