Summary
In this study I have used a tracer complex made of wheat germ agglutinin horseradish peroxidase conjugate (WGA*HRP) coupled to colloidal gold for retrograde tracing of neuronal pathways at the light microscopic level. Visualization of the gold was achieved by silver precipitation (the gold silver intensification method) with gold particles acting as specific cores of nucleation. The presence of horseradish peroxidase in the protein conjugate allowed this method to be compared with classical histochemistry using tetramethylbenzidine as a chromogen. The gold silver intensification method proved to be reliable, specific and sensitive. It has been demonstrated to be useful with fixatives containing a high percentage of paraformaldehyde and compatible with histochemical procedures to show projections of transmitter specific pathways.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Bowker RM, Westlund KN, Sullivan MC, Wilber JF, Coulter JD (1983) Descending scrotoninergic, peptidergic and cholinergic pathways from the raphe nuclei: a multiple transmitter complex. Brain Res 288:33–48
Danscher G (1981) Localization of gold in biological tissue. A photochemical method for light and electronmicroscopy. Histochemistry 71:81–88
Gallyas F (1982) Suppression of the argyrophil III reaction by mercapto compounds (A prerequisite for the intensification of certain histochemical reactions by physical developers). Acta Histochem 70:99–105
Geoghegan WD, Ackerman GA (1977) Adsorption of horseradish peroxidase, ovomucoid and anti-immunoglobulin to colloidal gold for the indirect detection of concanavalin A, wheat germ agglutinin and goat antihuman immunoglobulin G on cell surfaces at the electron microscopic level: a new method, theory and application. J Histochem 25:1187–1200
Gonatas NK, Harper C, Mizutani T, Gonatas JO (1979) Superior sensitivity of conjugates of horseradish peroxidase with wheat germ agglutinin for studies of retrograde axonal transport. J Histochem Cytochem 27:728–742
Goodman SL, Hodges GM, Trejdosiewicz LK, Livingston DC (1981) Colloidal gold markers and probes for routine application in microscopy. J Microsc 123:201–213
Hockfelt T, Terenius L, Kuypers HGJM, Dann O (1979) Evidence for enkephalin immunoreactive neurons in the medulla oblongata projecting to the spinal cord. Neurosci Lett 14:55–60
Hsu S, Raine L, Fanger H (1981) A comparative study of the peroxidase-antiperoxidase method and an avidin-biotin complex method for studying polypeptide hormones with radioimmunoassay antibodies. Am J Clin Pathol 75:734–738
Kristensson K, Olsson Y (1971) Retrograde axonal transport of protein. Brain Res 29:363–368
La Vail JH, La Vail MM (1972) Retrograde axonal transport in the central nervous system. Science 176:1416–1417
Lechan RM, Nestler JL, Jacobson S (1981) Immunohistochemical localization of retrogradely and anterogradely transported wheat germ agglutinin (WGA) within the central nervous system of the rat: application to immunostaining of a second antigen within the same neuron. J Histochem Cytochem 29:1255–1262
Lee CL, Menétrey D (1984) An electron microscopic method to demonstrate collateral projections in the central nervous system using gold-wheat germ agglutinin horseradish peroxidase conjugate. Neurosc Abs 10:420
MacLean IW, Nakane PK (1974) Periodate-lysine-paraformaldehyde fixative. A new fixative for immunoelectron microscopy. J Histochem Cytochem 22:1077–1083
Malmgren L, Olsson Y (1978) A sensitive method for histochemical demonstration of horseradish peroxidase in neurons following retrograde axonal transports. Brain Res 148:279–294
Mesulam MM (1982) Tracing neural connections with horseradish peroxidase. Wiley & Sons. New York, pp. 1–251
Muhlpfordi H (1982) The preparation of colloidal gold particles using tannic acid as an additional reducing agent. Experientia 38:1127–1128
Roberts WJ (1935) A new procedure for the detection of gold in animal tissue. Proc R Acad (Amsterdam) 38:540–544
Rye DB, Saper CB, Wayner BH (1984) Stabilization of the tetramethylbenzidine (TMB) reaction product: application for retrograde and anterograde tracing, and combination with immunohistochemistry. J Histochem Cytochem 32:1145–1153
Schwab ME, Thoenen H (1978) Selective binding, uptake and retrograde transport of tetanus toxin by nerve terminals in the rat iris. An electron microscope study using colloidal gold as a tracer. J Cell Biol 77:1–13
Sternberger LA, Hardy PH Jr, Cuculis JJ, Meyer HG (1970) The unlabeled antibody enzyme method of immunohistochemistry. Preparation and properties of soluble antigen-antibody complex (horseradish peroxidase- anti horseradish peroxidase) and its use in the identification of spirochetes. J Histochem Cytochem 18:315–331
Streefkerk JG (1972) Inhibition of erythrocyte pseudoperoxidase activity by treatment with hydrogen peroxide following methanol. J Histochem Cytochem 20:829–831
Zehr DR (1978) Use of hydrogen peroxide-egg albumin to eliminate nonspecific staining in immunoperoxidase techniques. J Histochem Cytochem 26:415–416
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Menétrey, D. Retrograde tracing of neural pathways with a protein-gold complex. Histochemistry 83, 391–395 (1985). https://doi.org/10.1007/BF00509197
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00509197