Abstract
The mammalian neocortex is a remarkable structure that is characterized by tangential surface expansion and six-layered lamination. However, how the mammalian neocortex emerged during evolution remains elusive. Because all modern reptiles have a homolog of the neocortex at the dorsal pallium, developmental analyses of the reptilian cortex are valuable to explore the origin of the neocortex. However, reptilian cortical development and the underlying molecular mechanisms remain unclear, mainly due to technical difficulties with sample collection and embryonic manipulation. Here, we introduce a method of embryonic manipulations for the Madagascar ground gecko and Chinese softshell turtle. We established in ovo electroporation and an ex ovo culture system to address neural stem cell dynamics, neuronal differentiation and migration. Applications of these techniques illuminate the developmental mechanisms underlying reptilian corticogenesis, which provides significant insight into the evolutionary steps of different types of cortex and the origin of the mammalian neocortex.
Highlights
The mammalian cerebral cortex is a remarkable brain structure that is responsible for intricate social behaviors and intelligence
Comparative analyses of extant amniote brains are powerful approaches to understand the evolutionary processes of the mammalian neocortex and homologous structures in non-mammalian lineages (Molnar et al, 2006; Aboitiz, 2011; Medina et al, 2013)
Previous histological studies revealed that the stellate morphology of migrating neurons in the developing reptilian cortex resemble migrating neurons in the early stages of mammalian neocortex (Goffinet, 1983)
Summary
The mammalian cerebral cortex is a remarkable brain structure that is responsible for intricate social behaviors and intelligence. We describe a method of embryonic manipulation techniques for two reptilian species: the Madagascar ground gecko (Paroedura pictus) and the Chinese softshell turtle (Pelodiscus sinensis). To allow embryonic development in the medium after electroporation, we established an ex ovo culture system for the middle stages of reptilian embryos (Figure 4 and Table 3).
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