Letter | Published:

Fluorescent peptides highlight peripheral nerves during surgery in mice

Nature Biotechnology volume 29, pages 352356 (2011) | Download Citation

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.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    Intraoperative facial nerve monitoring. Am. J. Otol. Nov., (Suppl.), 58–61 (1985).

  2. 2.

    , & Recurrent laryngeal nerve localization using a microlaryngeal electrode. Otolaryngol. Head Neck Surg. 87, 330–333 (1979).

  3. 3.

    & Identification and monitoring of the recurrent laryngeal nerve during thyroidectomy. Surg. Oncol. Clin. N. Am. 17, 121–144 (2008).

  4. 4.

    , & Basic principles of anatomy for optimal surgical treatment of prostate cancer. World J. Urol. 25, 31–38 (2007).

  5. 5.

    et al. A critical analysis of the current knowledge of surgical anatomy related to optimization of cancer control and preservation of continence and erection in candidates for radical prostatectomy. Eur. Urol. 57, 179–192 (2010).

  6. 6.

    et al. Impact of nerve sparing technique on patient self-assessed outcomes after radical perineal prostatectomy. J. Urol. 178, 488–492 (2007).

  7. 7.

    , , & Real-time mapping of the subepithelial nerve plexus by in vivo confocal laser scanning microscopy. Br. J. Ophthalmol. 94, 1133–1135 (2010).

  8. 8.

    , , , & Optical coherence tomography: a review of clinical development from bench to bedside. J. Biomed. Opt. 12, 051403 (2007).

  9. 9.

    et al. Currents concepts in neuroanatomical tracing. Prog. Neurobiol. 62, 327–351 (2000).

  10. 10.

    et al. Efficacy of seven retrograde tracers, compared in multiple-labelling studies of feline motoneurones. J. Neurosci. Methods 53, 35–46 (1994).

  11. 11.

    , , & In vivo visualization of the cochlear nerve and nuclei with fluorescent axonal tracers. Hear. Res. 162, 48–52 (2001).

  12. 12.

    et al. Fluorescent retrograde axonal tracing of the facial nerve. Laryngoscope 116, 1792–1797 (2006).

  13. 13.

    & Organ targeting in vivo using phage display peptide libraries. Nature 380, 364–366 (1996).

  14. 14.

    , , , & PepBank—a database of peptides based on sequence text mining and public peptide data sources. BMC Bioinformatics 8, 280 (2007).

  15. 15.

    , & Autoreactive antibodies are present in sheep with Johne's disease and cross-react with mycobacterium avium subsp. Paratuberculosis antigens. Microbes Infect. 9, 963–970 (2007).

  16. 16.

    et al. Suppression of receptor-mediated apoptosis by death effecter domain recruiting domain binding peptide aptamer. Biochem. Biophys. Res. Commun. 343, 1165–1170 (2006).

  17. 17.

    et al. Targeting of hepatoma cell and suppression of tumor growth by a novel 12mer peptide fused to superantigen TSST-1. Mol. Med. 12, 81–87 (2006).

  18. 18.

    et al. Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP. Neuron 28, 41–51 (2000).

  19. 19.

    et al. Activatable cell penetrating peptides attached to nanoparticles: dual probes for fluorescence and magnetic resonance imaging of proteases in vivo. Proc. Natl. Acad. Sci. USA 107, 4311–4316 (2010).

  20. 20.

    et al. Surgery with molecular fluorescence imaging using activatable cell penetrating peptides decreases residual cancer and improves survival. Proc. Natl. Acad. Sci. USA 107, 4317–4322 (2010).

  21. 21.

    , & Noninvasive model of sciatic nerve conduction in healthy and septic mice: reliability and normative data. Muscle Nerve 40, 610–616 (2009).

Download references

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.

Author information

Affiliations

  1. Department of Pharmacology, University of California at San Diego, La Jolla, California, USA.

    • Michael A Whitney
    • , Beth Friedman
    •  & Roger Y Tsien
  2. Department of Chemistry and Biochemistry, University of California at San Diego, La Jolla, California, USA.

    • Jessica L Crisp
    •  & Roger Y Tsien
  3. Division of Otolaryngology-Head and Neck Surgery, University of California at San Diego, La Jolla, California, USA.

    • Linda T Nguyen
    •  & Quyen T Nguyen
  4. Howard Hughes Medical Institute, University of California at San Diego, La Jolla, California, USA.

    • Larry A Gross
    • , Paul Steinbach
    •  & Roger Y Tsien

Authors

  1. Search for Michael A Whitney in:

  2. Search for Jessica L Crisp in:

  3. Search for Linda T Nguyen in:

  4. Search for Beth Friedman in:

  5. Search for Larry A Gross in:

  6. Search for Paul Steinbach in:

  7. Search for Roger Y Tsien in:

  8. Search for Quyen T Nguyen in:

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.

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.

Corresponding author

Correspondence to Quyen T Nguyen.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Table 1, Supplementary Figs. 1–8 and Supplementary Video 1

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nbt.1764