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

Amy M. Moore, Gregory H. Borschel, Katherine B. Santosa, Eric R. Flagg, Alice Y. Tong, Rahul Kasukurthi, Piyaraj Newton, Ying Yan, Daniel A. Hunter, Philip J. Johnson, Susan E. Mackinnon

Research output: Contribution to journalArticle

42 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)19-27
Number of pages9
JournalJournal of Neuroscience Methods
Volume204
Issue number1
DOIs
StatePublished - Feb 1 2012

Keywords

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

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