A transgenic rat expressing green fluorescent protein (GFP) in peripheral nerves provides a new hindlimb model for the study of nerve injury and regeneration

作者：Moore Amy M; Borschel Gregory H; Santosa Katherine A; Flagg Eric R; Tong Alice Y; Kasukurthi Rahul; Newton Piyaraj; Yan Ying; Hunter Daniel A; Johnson Philip J; Mackinnon Susan E^*

来源：Journal of Neuroscience Methods, 2012, 204(1): 19-27.

DOI：10.1016/j.jneumeth.2011.10.011

• Nerve regeneration
• Transgenic rat
• Green fluorescent protein
• Peripheral nerve
• Fluorescence microscopy
• Confocal microscopy

Background: In order to evaluate nerve regeneration in clinically relevant hindlimb surgical paradigms not feasible in fluorescent mice models, we developed a rat that expresses green fluorescent protein (GFP) in neural tissue.
Methods: Transgenic Sprague-Dawley rat lines were created using pronuclear injection of a transgene expressing GFP under the control of the thy1 gene. Nerves were imaged under fluorescence microscopy and muscles were imaged with confocal microscopy to determine GFP expression following sciatic nerve crush, transection and direct suturing, and transection followed by repair with a nerve isograft from nonexpressing littermates.
Results: In each surgical paradigm, fluorescence microscopy demonstrated the loss and reappearance of fluorescence with regeneration of axons following injury. Nerve regeneration was confirmed with imaging of Wallerian degeneration followed by reinnervation of extensor digitorum longus (EDL) muscle motor endplates using confocal microscopy.
Conclusion: The generation of a novel transgenic rat model expressing GFP in neural tissue allows in vivo imaging of nerve regeneration and visualization of motor endplate reinnervation. This rat provides a new model for studying peripheral nerve injury and regeneration over surgically relevant distances.

• 出版日期2012-2-15