Skip to main content

Tract Tracing Methods at the Light Microscopic Level

  • Chapter
  • 235 Accesses

Abstract

Tract tracing techniques provide tools for the study of the termination or origin of central neural pathways or peripheral nerves. The understanding of the organization of neural circuits has represented one of the major goals in neuroscience since its birth. In the second half of the 19th century, the pioneers in tract tracing discovered that retrograde degeneration of neuronal cell bodies and anterograde degeneration of fibers could be used to trace pathways in the nervous system. Thus, “the earliest way to identify the neurons sending their axons to a given neural structure was to destroy the structure” (20). In addition, the Golgi method, which impregnates random subsets of neuronal cell bodies and processes in their entirety, played a crucial role not only in unraveling the basic structure of the nervous system, but also in pioneering investigations on its connectivity. Cajal’s seminal studies on the wiring of the nervous system were in fact based on Golgi-impregnated material (5). It is very hard, however, to effectively impregnate the axons and to reconstruct their trajectory over long distances with the Golgi staining. This techconnections, i.e., at a limited distance from the cell body.

This is a preview of subscription content, log in via an institution.

Buying options

Chapter
USD   29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   129.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Alvarez-Buylla, A., C.Y. Ling, and J.R. Kirn. 1990. Cresyl violet: a red fluorescent Nissl stain. J. Neurosci. Methods 33:129–133.

    Article  PubMed  CAS  Google Scholar 

  2. Basbaum, A.I. and D. Menétrey. 1987. Wheat germ agglutinin-apoHRP gold: a new retrograde tracer for light-and electron-microscopic single-and double-label studies. J. Comp. Neurol. 261:306–318.

    Article  PubMed  CAS  Google Scholar 

  3. Bentivoglio, M. and S. Chen. 1993. Retrograde neuronal tracing combined with immunocytochemistry, p. 301–328. In A.C. Cuello (Ed.), IBRO Handbook Series: Methods in Neurosciences, Vol. 14, Immunohistochemistry II. John Wiley & Sons, Chichester.

    Google Scholar 

  4. Bentivoglio, M. and H.-S. Su. 1990. Photoconversion of fluorescent tracers. Neurosci. Lett. 113:127–133.

    Article  PubMed  CAS  Google Scholar 

  5. Cajal, S.R. 1911. Histologie du Système Nerveux de l’Homme et des Vertébrés. Maloine, Paris.

    Google Scholar 

  6. Callahan, C.A., S. Yoshikawa, and J.B. Thomas. 1998. Tracing axons. Curr. Opin. Neurobiol. 8:582–586.

    Article  PubMed  CAS  Google Scholar 

  7. Cowan, W.M., D.I. Gottlieb, A.E. Hendrickson, J.L. Price, and T.A. Woolsey. 1972. The autoradiographic demonstration of axonal connections in the central nervous system. Brain Res. 37:21–51.

    Article  PubMed  CAS  Google Scholar 

  8. Cuénod, M., P. Bagnoli, A. Beaudet, A. Rustioni, L. Wiklund, and P. Streit. 1982. Transmitter-specific retrograde labeling of neurons, p. 17–44. In V Chan-Palay and S. Palay (Eds.), Cytochemical Methods in Neuroanatomy. A.R. Liss, New York.

    Google Scholar 

  9. Fink, R.P. and L. Heimer. 1967. Two methods for selective silver impregnation of degenerating axons and their synaptic endings in the central nervous system. Brain Res. 4:369–374.

    Article  PubMed  CAS  Google Scholar 

  10. Gerfen, C.R., P.E. Sawchenko, and J. Carlsen. 1989. The PHA-L anterograde axonal tracing method, p. 19–47. In L. Heimer and L. Zaborsky (Eds.), Neuroanatomical Tract-Tracing Methods. Plenum Press, New York.

    Google Scholar 

  11. Glover, J.C. 1995. Retrograde and anterograde axonal tracing with fluorescent dextran-amines in the embryonic nervous system. Neurosci. Prot. 30:1–13.

    Google Scholar 

  12. Herrera, D.G. and H.A. Robertson. 1996. Activation of c-fos in the brain. Prog. Neurobiol. 50:83–107.

    Article  PubMed  CAS  Google Scholar 

  13. Honig, M.G. and R.I. Hume. 1986. Fluorescent carbocyanine dyes allow living neurons of identified origin to be studied in long-term cultures. J. Cell Biol. 103:171–187.

    Article  PubMed  CAS  Google Scholar 

  14. Honig, M.G. and R.I. Hume. 1989. Dil and DiO: versatile fluorescent dyes for neuronal labelling and pathway tracing. Trends Neurosci. 12:333–341.

    Article  PubMed  CAS  Google Scholar 

  15. Kristensson, K. and Y. Olsson. 1971. Retrograde axonal transport of protein. Brain Res. 29:363–365.

    Article  PubMed  CAS  Google Scholar 

  16. Kuypers, H.G. and G. Ugolini. 1990. Viruses as transneuronal tracers. Trends Neurosci. 13:71–75.

    Article  PubMed  CAS  Google Scholar 

  17. LaVail, J.H. and M.M. LaVail. 1972. Retrograde axonal transport in the central nervous system. Science 176:1416–1417.

    Article  PubMed  CAS  Google Scholar 

  18. Lübke, J. 1993. Photoconversion of different fluorescent substances for light and electron microscopy. Neurosci. Prot. 5:1–13.

    Google Scholar 

  19. Mesulam, M.M. 1978. Tetramethyl benzidine for horseradish peroxidase neurohistochemistry: a noncarcinogenic blue reaction product with superior sensitivity for visualizing neural afferents and efferents. J. Histochem. Cytochem. 26:106–117.

    PubMed  CAS  Google Scholar 

  20. Nauta, W.J.H. and M. Feirtag. 1986. Fundamental Neuroanatomy. Freeman & Co., New York.

    Google Scholar 

  21. Nauta, W.J.H. and P.A. Gygax. 1951. Silver impregnation of degenerating axon terminals in the central nervous system: (1) technic (2) chemical notes. Stain Technol. 26:3–9.

    Google Scholar 

  22. Olucha, F., G.F. Martinez, and G.C. Lopez. 1985. A new stabilizing agent for the tetramethyl benzidine (TMB) reaction product in the histochemical detection of horseradish peroxidase (HRP). J. Neurosci. Methods 13:131–138.

    Article  PubMed  CAS  Google Scholar 

  23. Schmued, L.C. 1994. Anterograde and retrograde neuroanatomical tract tracing with fluorescent compounds. Neurosci. Prot. 5:1–15.

    Google Scholar 

  24. Schmued, L.C. and L.F. Snavely. 1993. Photoconversion and electron microscopic localization of the fluorescent axon tracer fluoro-ruby (rhodamine-dextranamine). J. Histochem. Cytochem. 41:777–782.

    PubMed  CAS  Google Scholar 

  25. van der Want, J.J., J. Klooster, B.N. Cardozo, H. de Weerd, and R.S. Liem. 1997. Tract-tracing in the nervous system of vertebrates using horseradish peroxidase and its conjugates: tracers, chromogens and stabilization for light and electron microscopy. Brain Res. Brain Res. Protocol 1:269–279.

    Article  Google Scholar 

  26. Vercelli, A., M. Repici, D. Garbossa, and A. Grimaldi. 2000. Recent techniques for tracing pathways in the central nervous system of developing and adult mammals. Brain Res. Bull. 51:11–28.

    Article  PubMed  CAS  Google Scholar 

  27. Weinberg, R.J. and S.L. van Eyck. 1991. A tetramethylbenzidine/tungstate reaction for horseradish peroxidase histochemistry. J. Histochem. Cytochem. 39:1143–1148.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2002 Springer-Verlag New York, Inc.

About this chapter

Cite this chapter

Bentivoglio, M., Bertini, G. (2002). Tract Tracing Methods at the Light Microscopic Level. In: Merighi, A., Carmignoto, G. (eds) Cellular and Molecular Methods in Neuroscience Research. Springer, New York, NY. https://doi.org/10.1007/978-0-387-22460-2_12

Download citation

  • DOI: https://doi.org/10.1007/978-0-387-22460-2_12

  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-0-387-95386-1

  • Online ISBN: 978-0-387-22460-2

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics