Skip to main content
SpringerLink
Log in
Book cover

Neuroanatomical Tract-Tracing Methods pp 91–116Cite as

Methods for Delivering Tracers

  • Chapter

Abstract

The delivery of chemical tracers into the central nervous system is often an essential, although sometimes problematic aspect of modern tract-tracing experiments. In this chapter we shall concentrate on the most common methods of tracer delivery: small injections of a solution, either using a microsyringe or through glass micropipettes, or extracellular of intracellular iontophoresis. It should be kept in mind, however, that other methods of delivering tracers may be more suitable for certain types of problems. Tracers have been injected into the periphery—for instance, into muscle in order to label motoneurons in the spinal cord or brain stem (Kristenson, 1970; Kristenson et al., 1971; Kristenson and Olsson, 1973)—and they have been applied to the cut or crushed ends of peripheral nerves (Kristenson, 1975; lies and Mulloney, 1971; Tweedle, 1978) and deposited in naturally closed structures such as the eye (LaVail and LaVail, 1972, 1974). In cases where diffusion from the site of deposition is not a serious limitation, tracers may be applied directly to the surface of the brain, either as a liquid or by means of absorbant pellets or wicks that have been soaked in the tracer solution (Arbuthnott, 1969; Descarries and Lapierre, 1973; Held and Young, 1969; Weiss and Holland, 1967).

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

Chapter
USD   29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   84.99
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   109.99
Price excludes VAT (USA)
  • Compact, lightweight 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

  • Abols, 1. A., and Basbaum, A. L, 1979, The posterior pretectal nucleus: Evidence for a direct projection to the inferior olive of the cat, Neurosci. Lett. 13: 111–116.

    Article  Google Scholar 

  • Aghajanian, G. K., and Wang, R. Y., 1977, Habenular and other midbrain afferents demonstrated by a modified retrograde tracing technique. Brain Res. 122: 229–242.

    Article  PubMed  CAS  Google Scholar 

  • Ahlsen, G., Lindstrom, S., and Sybirska, E., 1978, Subcortical axon collaterals of principal cells in the lateral geniculate body of the cat. Brain Res. 156: 106–109.

    Article  PubMed  CAS  Google Scholar 

  • Arbuthnott, G. W., 1969, Noradrenaline uptake into cerebral cortex: A histochemical study, J. Neurochem. 16: 1599–1604.

    Article  CAS  Google Scholar 

  • Barrett, J., and Whitlock, D. G., 1973, Technique for believing glass microelectrodes, in: Intracellular Staining in Neurobiology ( S. B. Kater and C. Nicholsen, eds.), pp. 297–299, Springer-Verlag, Berlin.

    Google Scholar 

  • Beckstead, R. M., 1979a, An autoradiographic examination of corticocortical and subcortical projections of the mediodorsal projection (prefrontal) cortex in the rat, J. Comp. Neurol. 184: 43–62.

    Article  PubMed  CAS  Google Scholar 

  • Beckstead, R. M., 1979b, Convergent prefrontal and nigral projections to the striatum of the rat, Neurosci. Lett. 123: 59–64.

    Article  Google Scholar 

  • Brown, K. T., and Flaming, D. G., 1974, Beveling of fine micro pipette electrodes by a rapid precision method, Science 185: 693–695.

    Article  PubMed  CAS  Google Scholar 

  • Brown, K. T., and Flaming, D. G., 1975, Instrumentation and technique for beveling fine micropipette electrodes. Brain Res. 86: 172–180.

    Article  PubMed  CAS  Google Scholar 

  • Brown, K. T., and Flaming, D. G., 1977, New microelectrode techniques for intracellular work in small cells, Neuroscience 2: 813–827.

    Article  Google Scholar 

  • Brown, K. T., and Flaming, D. G. 1979, Technique for precision beveling of relatively large micropipettes, J. Neurosci. Methods 1: 25–34.

    Article  CAS  Google Scholar 

  • Brown, P. B., Maxfield, B. W., and Moraff, H., 1973, Electronics for Neurobiologists, M.I.T. Press, Cambridge, Massachusetts.

    Google Scholar 

  • Bunney, B. S., and Aghajanian, G. K., 1976, The precise localization of nigral afferents in the rat as determined by a retrograde tracing technique. Brain Res. 117: 423–435.

    Article  PubMed  CAS  Google Scholar 

  • Bunt, A., and Haschke, R. H., 1978, Features of foreign proteins affecting their retrograde transport in axons of the visual system, J. Neurocytol. 7: 665–678.

    Article  CAS  Google Scholar 

  • Bunt, A. H., Haschke, R. H., Lund, R. D., and Calkins, D. F., 1976, Factors affecting retrograde axonal transport of horseradish peroxidase in the visual system, Brain Res. 102: 152–155.

    Article  PubMed  CAS  Google Scholar 

  • Bures, J., Buresova, O., and Huston, J., 1976, Techniques and Basic Experiments for the Study of Brain and Behavior, Elsevier, Amsterdam.

    Google Scholar 

  • Cedarbaum, J. M., and Aghajanian, G. K., 1978, Afferent projections to the rat locus coeruleus as determined by a retrograde tracing technique, J. Comp. Neurol. 178: 1–16.

    Article  CAS  Google Scholar 

  • Chowdhury, T. K., 1969, Techniques of intracellular injection, in Glass Microelectrodes ( M. La Vallee, O. F. Schanne, and N. C. Herbert, eds.), pp. 404–423, John Wiley & Sons, New York.

    Google Scholar 

  • Corson, D. W., Goodman, S., and Fein, A., 1979, An adaptation of the jet stream micropipette beveler, Science 205: 1302.

    Article  PubMed  CAS  Google Scholar 

  • Davis, R. E., and Agranoff, B. W., 1977, Microimplantation of 3H-proline on a single bead of ion-exchange resin, Brain Res. 124: 341–346.

    Article  PubMed  CAS  Google Scholar 

  • Descarries, L., and Lapierre, Y., 1973, Noradrenergic axon terminals in the cerebral cortex of rat. I. Radioautographic visualization after topical application of d, 1 3H-norepinephrine, Brain Res. 51: 141–160.

    Article  PubMed  CAS  Google Scholar 

  • Dreyer, F., and Peper, K., 1974, lontophoretic applications of acetylcholine: Advantages of high resistance micropipettes in connection with an electronic current pump, Pfluegers Arch. 348: 263–272.

    Google Scholar 

  • Edwards, S. B., and Henkel, C. K., 1978, Superior colliculus connections with the extraocular motor nuclei in the cat, J. Comp. Neurol. 179: 451–468.

    Article  CAS  Google Scholar 

  • Edwards, S. B., and Shalna, E. J., 1974, Microinjector for use in the autoradiographic neuroanatomical tracing method, Pharmacol. Biochem. Behav. 2: 111–113.

    Article  PubMed  CAS  Google Scholar 

  • Eide, E., Illert, M., and Tanaka, R., 1976, Injection of horseradish peroxidase solution in calibrated volumes (nanolitre) into the spinal cord, Neurosci. Lett. 2: 51–56.

    Article  PubMed  CAS  Google Scholar 

  • Fein, H., 1966, Passing current through recording glass micro–pipette electrodes, IEEE Trans. Biomed. Eng. 13: 211–212.

    Article  PubMed  CAS  Google Scholar 

  • Gallager, D. W., and Pert, A., 1978, Afferents to brain stem nuclei (brain stem raphe, nucleus reticularis pontis caudalis and nucleus giganto cellularis) in the rat as demonstrated by microiontophoretically applied horseradish peroxidase. Brain Res. 144: 257–275.

    Article  PubMed  CAS  Google Scholar 

  • Geller, H. M., and Woodward, D. J., 1972, An improved constant current source for micro-iontophoretic drug application studies, Electroencephalogr. Clin. Neurophysiol. 33: 430–432.

    Article  PubMed  CAS  Google Scholar 

  • Graybiel, A. M., and Devor, M. A., 1974, A microelectrode delivery technique for use with horseradish peroxidase, Brain Res. 68: 167–173.

    Article  PubMed  CAS  Google Scholar 

  • Griffin, G., Watkins, L. R., and Mayer, D. V., 1979, HRP pellets and slow release gels: Two new techniques for greater localization and sensitivity. Brain Res. 168: 595–601.

    Article  PubMed  CAS  Google Scholar 

  • Guglielmotti, V., 1979, A thermally controlled microinjector, Stain Technol. 54: 151–157.

    PubMed  CAS  Google Scholar 

  • Held, J., and Young, I. J., 1969, A comparative study of the somato-axonal flow of protein in the feline hypoglossal and vagus nerves, Exp. Brain Res. 8: 150–162.

    CAS  Google Scholar 

  • Henkel, C. K., and Edwards, S. B., 1978, The superior colliculus control of pinna movements in the cat: Possible anatomical connections, J. Comp. Neurol. 182: 763–776.

    Article  CAS  Google Scholar 

  • Herkenham, M., 1978, The connections of the nucleus reuniens thalami: Evidence for a direct thalamo–hippocampal pathway in the rat, J. Comp. Neurol. 177: 589–610.

    Article  CAS  Google Scholar 

  • Herkenham, M., 1979, The afferent and efferent connections of the ventromedial thalamic nucleus in the rat, J. Comp. Neurol. 183: 487–518.

    Article  CAS  Google Scholar 

  • Herkenham, M., and Nauta, W. J. H., 1979, Efferent connections of the habenula nuclei in the rat, J. Comp. Neurol. 187: 19–48.

    Article  CAS  Google Scholar 

  • Hoenig, S. A., and F. L. Payne, 1973, How to Build and Use Electronic Devices without Frustration, Panic, Mountains of Money, or an Engineering Degree, Little, Brown & Co., Boston.

    Google Scholar 

  • Holland, V. R., Saunders, B. C., Rose, F. L., and Walpole, A. L., 1974, A safer substitute for benzidine in detection of blood. Tetrahedron 30: 3299–3302.

    Article  CAS  Google Scholar 

  • Iles, J. F., and Mulloney, B., 1971, Procion yellow staining of cockroach motor neurons without the use of microelectrodes, Brain Res. 30: 397–400.

    Article  PubMed  CAS  Google Scholar 

  • Jones, E. G., Coulter, J. D., and Hendry, S. H. C., 1978, Intracortical connectivity of architectonic fields in the somatic sensory, motor and parietal cortex of monkeys, J. Comp. Neurol 181: 291–348.

    Article  CAS  Google Scholar 

  • Kallstrom, Y., and Lindstrom, S., 1978, Simple device for pressure injections of horseradish peroxidase into small central neurons. Brain Res. 156: 102–105.

    Article  PubMed  CAS  Google Scholar 

  • Kater, S. B., and Nicholson, C., 1973, Intracellular Staining in Neurobiology, Springer-Verlag, Berlin and New York.

    Google Scholar 

  • Kater, S., Nicholson, C., and Davis, W. J., 1973, Guide to intracellular staining techniques, in: Intracellular Staining in Neurobiology ( S. B. Kater and C. Nicholson, eds.), pp. 307–325, Springer-Verlag, Berlin and New York.

    Google Scholar 

  • Kawamura, S., and Diamond, I. T., 1978, The laminar origin of descending projections from the cortex to the thalamus in Topaia glis, Brain Res. 153: 333–339.

    Article  PubMed  CAS  Google Scholar 

  • Kawamura, S., Fukushima, N., Hattori, S., and Tashiro, T., 1978, A ventral lateral geniculate nucleus projection to the dorsal thalamus and the midbrain in the cat, Exp. Brain Res. 34: 95–106.

    Google Scholar 

  • Kelly, J. P., and Gilbert, C. D., 1975, The projections of different morphological types of ganglion cells in the cat retina, J. Comp. Neurol 163: 65–80.

    Article  CAS  Google Scholar 

  • Koike, H., Eisenstadt, M., and Schwartz, J. H., 1972, Axonal transport of newly synthesized acetylcholine in an identified neuron of aplysia, Res. 37: 152–159.

    CAS  Google Scholar 

  • Kristenson, K., 1970, Transport of fluorescent protein tracer in peripheral nerves. Acta Neuropathol (Berlin) 16: 293–300.

    Article  Google Scholar 

  • Kristenson, K., 1975, Retrograde axonal transport of protein tracers, in: The Use of Axonal Transport for Studies of Neuronal Connectivity ( W. M. Cowan and M. Cuenod, eds.), Elsevier, Amsterdam.

    Google Scholar 

  • Kristenson, K., and Olsson, Y., 1973, Diffusion pathways and retrograde axonal transport of protein tracers in peripheral nerves. Prog. Neurobiol. 1: 85–109.

    Article  Google Scholar 

  • Kristenson, K., Olsson, Y., and Sjostrand, J., 1971, Axonal uptake and retrograde transport of exogenous proteins in the hypoglossal nerve. Brain Res. 32: 399–406.

    Article  Google Scholar 

  • Lasek, R., Joeseph, B. S., and Whitlock, D. G., 1968, Evaluation of a radiautographic neuroanatomical tracing method. Brain Res. 8: 319–336.

    Article  PubMed  CAS  Google Scholar 

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

    Article  Google Scholar 

  • La Vail, J. H., and La Vail, M. M., 1974, The retrograde intraaxonal transport of horseradish peroxidase in the check visual system: A light and electron microscope study, J. Comp. Neurol. 157: 303–358.

    Article  Google Scholar 

  • La Vallee, M., Schanne, O. F., and Herbert, N. C., 1969, Glass Microelectrodes, John Wiley & Sons, New York.

    Google Scholar 

  • Luiten, P. G. M., 1975, The horseradish peroxidase technique applied to the teleostan nervous system, Brain Res. 89: 181–186.

    Article  PubMed  CAS  Google Scholar 

  • Luiten, P. G. M., and Van der Pers, J. N. C., 1977, The connection of the trigeminal and facial motor nuclei in the brain stem of the carp (Cyprinus carpio 1.) as revealed by anterograde and retrograde transport of horseradish peroxidase, J. Comp. Neurol. 174: 575–590.

    Article  CAS  Google Scholar 

  • Lynch, G., Deadwyler, S., and Gall, C., 1974, Labeling of central nervous system neurons with extracellular recording microelectrodes. Brain Res. 66: 337–341.

    Article  Google Scholar 

  • Maekawa, K., and Takeda, T., 1979, Origin of descending afferents to the rostral part of the dorsal cap of inferior olive which transfers contralateral optic activities to the flocculus. A horseradish peroxidase study, Brain Res. 172: 393–405.

    Article  PubMed  CAS  Google Scholar 

  • McCaman, R. E., McKenna, D. G., and Ono, J. K., 1977, A pressure system for intracellular and extracellular ejections of picoliter volumes. Brain Res. 136: 141–147.

    Article  PubMed  CAS  Google Scholar 

  • Myers, R. D., 1974, Handbook of Drug and Chemical Stimulation of the Brain, Van Nostrand Reinhold Co., London.

    Google Scholar 

  • Nastuk, W. L., 1964, Physical Techniques in Biological Research, Vol. V. Electrophysiological Methods, Part A, Academic Press, New York.

    Google Scholar 

  • Norgren, R., 1976, Taste pathways to hypothalamus and amygdala, J. Comp. Neurol. 166: 17–30.

    Article  CAS  Google Scholar 

  • Ogden, T. E., Citron, M. C., and Pierantoni, R., 1978, The jet stream microbeveler: An inexpensive way to bevel ultrafine glass micropipettes. Science 201: 469.

    Article  PubMed  CAS  Google Scholar 

  • Ottersen, O. P., and Ben-Ari, Y., 1978a, Pontine and mesencephalic afferents to the central nucleus of the amygdala of the rat, Neurosci. Lett. 8: 329–334.

    Article  PubMed  CAS  Google Scholar 

  • Ottersen, O. P., and Ben-Ari, Y., 1978b, Demonstration of a heavy projection of midline thalamic neurons upon the lateral nucleus of the amygadala of the rat, Neurosci. Lett. 9: 147–152.

    Article  PubMed  CAS  Google Scholar 

  • Ottersen, O. P., and Ben-Ari, Y., 1979, Afferent connections to the amygdaloid complex of the rat and cat. 1. Projections from the thalamus, J. Comp. Neurol. 187: 401–424.

    Article  CAS  Google Scholar 

  • Phillipson, O. T., 1978, Afferent projections to A10 dopaminergic neurons in the rat as shown by the retrograde transport of horseradish peroxidase, Neurosci. Lett. 9: 353–359.

    Article  PubMed  CAS  Google Scholar 

  • Phillipson, O. T., 1979, Afferent projection to the ventral tegmental area of Tsai and interfascicular nucleus: A horseradish peroxidase study in the rat, J. Comp. Neurol. 187: 117–144.

    Article  CAS  Google Scholar 

  • Pitman, R. M., Tweedle, C. D., and Cohen, M. J., 1972, Branching of central neurons: Intracellular cobalt injections for light and electron microscopy. Science 176: 412–414.

    Article  PubMed  CAS  Google Scholar 

  • Price, P., Fisher, A. W. F., and Redstone, P., 1977, A simple apparatus for injection of small (nanolitre) volumes of horseradish-peroxidase, Neurosci. Lett. 6: 21–25.

    Article  PubMed  CAS  Google Scholar 

  • Remler, M. P., Selverston, A. I., and Kennedy, D., 1968, Lateral giant fibers of crayfish: Location of somata by dye injection. Science 162: 281–283.

    Article  PubMed  CAS  Google Scholar 

  • RoBards, M. J., 1979, Somatic neurons in the brainstem and neocortex projecting to the external nucleus of the inferior colliculus: An anatomical study in the opossum, J. Comp. Neurol. 184: 547–566.

    Article  CAS  Google Scholar 

  • Sakai, M., Sakai, H., and Woody, C. D., 1978, Intracellular staining of cortical neurons by pressure microinjections and recovery by core biopsy, Exp. Neurol. 58: 138–144.

    Article  CAS  Google Scholar 

  • Schubert, P., and Holländer, H., 1975, Methods for delivery of tracers to the central nervous system, in: The Use of Axonal Transport for Studies of Neuronal Connectivity ( W. M. Cowan and M. Cuenod, eds.), Elsevier, Amsterdam.

    Google Scholar 

  • Schubert, P., Kreutzberg, G. W., and Lux, H. D., 1972, Use of microelectrophoresis in the autoradiographic demonstration of fiber projections. Brain Res. 39: 274–277.

    Article  PubMed  CAS  Google Scholar 

  • Shaw, M. L., and Lee, D. R., 1973, Micropipette sharpener with audio and hydraulic readouts, J. Appl. Physiol. 34: 523–524.

    PubMed  CAS  Google Scholar 

  • Simon, A., LeMoal, M., and Galas, A., 1979, Efferents and afferents of the ventral tegmental A-10 region studied after local injection of ®H-leucine and horseradish peroxidase, Brain Res. 178: 17–40.

    Article  PubMed  CAS  Google Scholar 

  • Spencer, H. J., Jones, R. K., and Lynch, G., 1978, The use of somatougal transport of horseradish peroxidase for tract tracing and cell labeling, in: Methods in Physiological Psychology, Vol. II, Neuroanatomical Research Techniques ( R. T. Robertson, ed.). Academic Press, New York and London.

    Google Scholar 

  • Tasaki, I., Tsukahara, Y., Ito, S., Wayner, M. J., and Yu, W. Y., 1968, A simple, direct and rapid method for filling microelectrodes, Physiol. Behav. 3: 1009–1010.

    Article  Google Scholar 

  • Thompson, R. F., and Patterson, M. M., 1973, Methods in Physiological Psychology, Vol. 1. Bioelectric Recording Techniques, Part A, Cellular Processes and Brain Potentials, Academic Press, New York and London.

    Google Scholar 

  • Thompson, R. F., and Patterson, M. M., 1974, Methods in Physiological Psychology, Vol. I. Bioelectric Recording Techniques, Part C, Receptor and Effector Processes, Academic Press, New York and London.

    Google Scholar 

  • Tweedle, G. D., 1978, Single cell staining techniques, in: Methods in Physiological Psychology, Vol. II, Neuroanatomical Research Techniques ( R. T. Robertson, ed.). Academic Press, New York and London.

    Google Scholar 

  • Wakefield, G., and Shonnard, N., 1979, Observations of HRP labeling following injection through a chronically implanted cannula. A method to avoid diffusion into injured fibers. Brain Res. 168: 221–226.

    Article  PubMed  CAS  Google Scholar 

  • Weiss, P., and Holland, Y., 1967, Neuronal dynamics and axonal flow. II. The olfactory nerve as a model test object, Proc. Natl. Acad. Sei. USA 57: 259–264.

    Google Scholar 

  • Woodward, W. R., and Lindström, S. H., 1977, A potential screening technique for neurotransmitters in the GNS: Model studies in the cat spinal cord. Brain Res. 137: 37–52.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Neurology, University of Virginia School of Medicine, Charlottesville, Virginia, 22908, USA

    George F. Alheid

  2. Department of Anatomy, University of Virginia School of Medicine, Charlottesville, Virginia, 22908, USA

    Stephen B. Edwards

  3. Department of Anatomy, Michigan State University, East Lansing, Michigan, 48824, USA

    Stephen T. Kitai & Melburn R. Park

  4. Comparative Animal Research Laboratory, Oak Ridge, Tennessee, 37830, USA

    Robert C. Switzer III

Authors
  1. George F. Alheid
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Stephen B. Edwards
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Stephen T. Kitai
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Melburn R. Park
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Robert C. Switzer III
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Department of Neurology, University of Virginia School of Medicine, Charlottesville, Virginia, 22908, USA

    Lennart Heimer

  2. University of California, Irvine, California, USA

    Martine J. Robards

Rights and permissions

Reprints and permissions

Copyright information

© 1981 Plenum Press, New York

About this chapter

Cite this chapter

Alheid, G.F., Edwards, S.B., Kitai, S.T., Park, M.R., Switzer, R.C. (1981). Methods for Delivering Tracers. In: Heimer, L., Robards, M.J. (eds) Neuroanatomical Tract-Tracing Methods. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-3189-6_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-4613-3189-6_3

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4613-3191-9

  • Online ISBN: 978-1-4613-3189-6

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation