A new method to study nerve fiber projecting in the spinal cord during chicken embryo development

doi:10.3969/j.issn.0529-1356.2014.05.025

Abstract

Abstract:

Objective To develop a method of studying fiber projecting in the spinal cord duiring chicken embryo development. Methods At embryonic incubation 3 day (E3), pCAGGS-green fluorescent protein (GFP) plasmid was injected into the spinal cord usingin vivo electroporation. Three days after transfection (E6), GFP-positive embryos were collected under a stereo fluorescence microscope. Subsequently, the spinal cord was separated from the embryos and cut from the roof plate as an open book. After fixed with 4% paraformaldehyde (PFA) for one hour, the opened spinal cords were used for immunohistochemistry with N-cadherin antibody and with DAPI for nuclei. Finally, the nerve fiber projecting was photographed and analyzed under a fluorescence microscope. Results Based on the opened spinal cord and immunostaining in the cryosection, we observed that the nerve fibers projected across the midline of the floor plate and reached to the sulcus terminalis along the white matter of the contra side. The immunoreaction against N-cadherin indicated that overexpression of GFP has no significant effect on chicken embryonic development. Conclusion A new method to study fiber projecting in the developing chicken spinal cord is established successfully in this study.

Key words: Spinal cord, Nerve fiber, Projecting, Electroporationin vivo, Chicken embryo

HU A-zhen1 YANG Ci-qing FU SU-lei JIA Yang-yang LI Han GUO Zhi-kun LIN Jun-tang *. A new method to study nerve fiber projecting in the spinal cord during chicken embryo development[J]. AAS, 2014, 45(5): 724-728.

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