Quantitative and qualitative analysis of Wallerian degeneration using restricted axonal labelling in YFP-H mice.

Abstract

We investigated the usefulness of YFP-H transgenic mice [Neuron 28 (2000) 41] which express yellow fluorescent protein (YFP) in a restricted subset of neurons to study Wallerian degeneration in the PNS. Quantification of YFP positive axons and myelin basic protein (MBP) immunocytochemistry revealed that YFP was randomly distributed to approximately 3% of myelinated motor and sensory fibres. Axotomy-induced Wallerian degeneration appeared as fragmentation of fluorescent signals in individual YFP positive axons with a morphology and timing similar to Wallerian degeneration observed by more traditional methods. In YFP-H transgenic mice co-expressing a high dosage of WldS, a chimeric gene that protects from Wallerian degeneration [Nat Neurosci. 4 (2001) 1199], axonal fragmentation in distal tibial nerves after sciatic nerve axotomy was approximately 10 times delayed. Considerable retardations of Wallerian degeneration using the same transgenic expression system were also observed in cultures of nerve explants, enabling in vitro real-time imaging of axonal fragmentation. Remarkably, single YFP-labelled axons could be traced in peripheral nerves for unusually long distances of up to 2.9 cm exploiting confocal fluorescence imaging. Altogether transgenic YFP-H mice prove to be a valuable tool to study mechanisms of Wallerian degeneration in vivo and in vitro.

• Authors: Bogdan Beirowski, Livia Berek, Robert Adalbert, Diana Wagner, Daniela S Grumme, Klaus Addicks, Richard R Ribchester, Michael P Coleman
• Journal: Journal of neuroscience methods (J Neurosci Methods) Vol. 134 Issue 1 Pg. 23-35 (Mar 15 2004) ISSN: 0165-0270 [Print] Netherlands
• PMID: 15102500 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Bacterial Proteins
• Luminescent Proteins
• yellow fluorescent protein, Bacteria

Topics

• Animals
• Axons (chemistry, ultrastructure)
• Bacterial Proteins (biosynthesis, genetics)
• Luminescent Proteins (biosynthesis, genetics)
• Mice
• Mice, Transgenic
• Sciatic Neuropathy (genetics, pathology)
• Staining and Labeling (methods)
• Wallerian Degeneration (genetics, pathology)