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A combination of both in vivo and in vitro techniques is invaluable for studying semaphorin signaling in the avian central nervous system. Here we describe how both types of approaches can be used to compliment each other in order to unravel the role that semaphorins play during embryonic development and elucidate the functional consequences of semaphorin knockdown using RNA interference vectors. We describe and discuss specifically the use of in ovo electroporation and primary oculomotor neuron culture to identify the role of semaphorins in oculomotor neuron migration and assess functional consequences of semaphorin disruption in this system.

Reptiles and birds have been highlighted as excellent experimental models for the study of developmental biology; however, due to technical limitations in cellular analysis, dynamics of neural stem/progenitor cells of these animals remain unclear. In this chapter, we introduce the protocols for neurosphere culture and ex ovo embryonic culture of developing reptilian and avian embryos, which are modified from the method originally established for rodent embryos. Applications of these techniques provide powerful strategies for the study of comparative neural development of amniotes.

The avian embryo has proven utility for studying ethanol's damaging effects upon the embryo. Chicken and quail are long-established models for developmental biology research; much of what we know regarding limb, craniofacial, neural crest, hindbrain, and cardiac morphogenesis was first established with avian models. These models also are for popular mechanistic studies of teratogens, including ethanol. Avian models have been used to explore ethanol's effects on neurogenesis, cardiogenesis, intracellular signaling, neurobehavior, and apoptosis. Presented here are several of these methodologies for adaptation by interested researchers.