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Fluorescent peptides highlight peripheral nerves during surgery in mice

Abstract

Nerve preservation is an important goal during surgery because accidental transection or injury leads to significant morbidity, including numbness, pain, weakness or paralysis. Nerves are usually identified by their appearance and relationship to nearby structures or detected by local electrical stimulation (electromyography), but thin or buried nerves are sometimes overlooked. Here, we use phage display to select a peptide that binds preferentially to nerves. After systemic injection of a fluorescently labeled version of the peptide in mice, all peripheral nerves are clearly delineated within 2 h. Contrast between nerve and adjacent tissue is up to tenfold, and useful contrast lasts up to 8 h. No changes in behavior or activity are observed after treatment, indicating a lack of obvious toxicity. The fluorescent probe also labels nerves in human tissue samples. Fluorescence highlighting is independent of axonal integrity, suggesting that the probe could facilitate surgical repair of injured nerves and help prevent accidental transection.

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Figure 1: Whole-body survey of nerves in mice (n = 3) 4 h after injection with 450 nmoles of FAM-NP41.
Figure 2: Time course and dose response of FAM-NP41 binding to nerve and nonnerve tissue.
Figure 3: Cy5-NP41 (acetyl-SHSNTQTLAKAPEHTGC-(Cy5)-amide) labeling of sciatic nerve in Thy1-YFP transgenic mice.
Figure 4: NP41 can highlight buried nerve branches invisible by standard illumination.

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Acknowledgements

We are indebted to members of our laboratory for discussions and comments on the manuscript. Results described here are being used in support of a patent filing by the University of California, San Diego. This work was supported by the Howard Hughes Medical Institute, grants from the Burrough-Wellcome Fund (Career Award for Medical Scientists) and National Institutes of Health (NIH, 5K08EB008122) to Q.T.N. and NIH grant NS27177 to R.Y.T.

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Authors

Contributions

M.A.W. designed and performed experiments, interpreted data and wrote manuscript. J.L.C. designed and performed experiments and interpreted data. L.T.N. designed and performed experiments and interpreted data. B.F. designed and performed experiments and interpreted data. L.A.G. designed and performed experiments and interpreted data. P.S. provided computer support for experiments, R.Y.T. designed experiments, interpreted data and wrote manuscript. Q.T.N. designed and performed experiments, interpreted data and wrote manuscript.

Corresponding author

Correspondence to Quyen T Nguyen.

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Competing interests

M.A.W., R.Y.T. and Q.T.N. are scientific advisors for Avelas Biosciences, Inc., who has licensed the technology described above from the University of California San Diego.

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Supplementary Table 1, Supplementary Figs. 1–8 and Supplementary Video 1 (PDF 12353 kb)

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Whitney, M., Crisp, J., Nguyen, L. et al. Fluorescent peptides highlight peripheral nerves during surgery in mice. Nat Biotechnol 29, 352–356 (2011). https://doi.org/10.1038/nbt.1764

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