Live imaging of neuroepithelial cells in the rat spinal cord by confocal laser-scanning microscopy

The vertebrate nervous system consists of a huge number of neurons and glial cells. These cell types are generated from neuroepithelial cells at precise positions during development. However, little is known about how kinetics of neuroepithelial cells is regulated, and how the balance of proliferation and differentiation are genetically coordinated. To elucidate cellular mechanisms underlying such complex cell behaviors in the neuroepithelium, we established time-lapse imaging system by applying confocal laser-scanning microscopy for the rat spinal cord slice culture. By electroporation the spinal cord with expression plasmids of fluorescent proteins, nuclear movement of neuroepithelial cells and morphological change on their radial fibers were clearly visualized. Four-dimensional (4-D) time-lapse analyses allowed us to investigate the spatiotemporal dynamics of neuroepithelial cells. These approaches make it possible to analyze functions of genes that regulate cell kinetics and cell morphology.