Summary
Evans blue bound to albumin, forming a vividly red fluorescent complex, was injected into the gastrocnemius muscle of suckling and adult mice. Its distribution in the peripheral and central nervous system was studied. The principal findings were as follows:
Marked age dependent differences were observed. In the adult mice a red fluorescence was found in the sciatic nerve restricted to the epi- and perineurium, and there was no staining of the endoneurium. In suckling mice, on the other hand, a red fluorescent material was present also in the endoneurium of the sciatic nerve.
In suckling mice red fluorescent granules in the cytoplasm of the ipsilateral motor neurones of the spinal cord were found. These motor neurones, showing a red fluorescence were restricted to a level corresponding to the entrance of the spinal roots of the sciatic nerve into the spinal cord. In adult mice no such fluorescence was found in the corresponding motor neurones.
This finding is considered to provide a convincing evidence that transport of a substance can take place in peripheral nerves in a disto-proximal direction. Certain similarities to the behaviour of herpes simplex virus infection after peripheral inoculation of virus, as regards the affection of the motor neurones and the differences between suckling and adult mice, are pointed out.
Zusammenfassung
An Albumin gebundenes Evansblau, das einen Komplex mit lebhafter Rotfluorescenz ergibt, wurde in den M. gastrocnemicus von Saug- und erwachsenen Mäusen injiziert. Seine Verteilung im peripheren und zentralen Nervensystem wurde untersucht. Die wichtigsten Befunde sind:
Deutliche altersabhängige Unterschiede wurden angetroffen. Bei erwachsenen Mäusen blieb die Rotfluorescenz auf das Epi- und Perineurium beschränkt und es fand sich keine Anfärbung des Endoneurium. Bei Säugmäusen hingegen war rot fluorescierendes Material auch im Endoneurium des N. ischiadicus vorhanden.
Bei Saugmäusen fanden sich rot fluorescierende Granula im Cytoplasma homolateraler Motoneurone des Rückenmarks. Die Fluorescenz der Nervenzellen war auf das dem Eintritt der Spinalwurzeln des N. ischiadicus entsprechende Rückenmarkssegment beschränkt. Bei adulten Mäusen war eine solche Fluorescenz entsprechender Motoneurone nicht nachweisbar.
Diese Befunde ergeben einen überzeugenden Hinweis dafür, daß in peripheren Nerven ein Substanztransport in disto-proximaler Richtung erfolgen kann. Auf gewisse Ähnlichkeiten mit dem Verhalten des Herpes simplex Virus nach peripherer Virusapplikation im Hinblick auf die Affektion der spinalen Motoneurone sowie die Unterschiede zwischen Saugmäusen und erwachsenen Tieren wird hingewiesen.
Similar content being viewed by others
References
Andervont, H. B.: Activity of herpetic virus in mice. J. infect. Dis.44, 383–393 (1929).
Becker, N. H., Hirano, A., Zimmerman, H. M.: Observations of the distribution of exogenous peroxidase in the rat cerebellum. J. Neuropath. exp. Neurol.27, 439–452 (1968).
Brightman, M. W.: The intracerebral movement of proteins injected into blood and cerebrospinal fluid of mice. Progr. Brain Res.29, 19–37 (1968).
Clasen, R. A., Pandolfi, S., Hass, G. M.: Vital staining, serum albmuin and the blood-brain barrier. J. Neuropath. exp. Neurol.29, 266–284 (1970).
Dahlström, A.: Observations on the accumulation of noradrenaline in the proximal and distal parts of peripheral adrenergic nerves after compression. J. Anat. (Lond.)99, 677–689 (1965).
Hamberger, A., Hamberger, B.: Uptake of catecholamines and penetration of trypan blue after blood-brain lesions. Z. Zellforsch.70, 386–392 (1966).
Holtzman, E., Peterson, E.: Cytochemical studies of protein uptake, lysosomes and GERL in neurons. J. Histochem. Cytochem.16, 503 (1968).
Hughes, A.: The growth of embryonic neurites. A study on cultures of chick neural tissue. J. Anat. (Lond.)87, 150–162 (1953).
Johnson, R. T.: The pathogenesis of herpes virus encephalitis. II. A cellular basis for the development of resistance with age. J. exp. Med.120, 359–374 (1964).
Kapeller, K., Mayor, D.: An electron microscopic study of the early changes distal to a constriction in sympathetic nerves. Proc. roy. Soc. B172, 53–63 (1969).
Kristensson, K.: Morphological studies of the neural spread of herpes simplex virus to the central nervous system. Acta neuropath. (Berl.)16, 54–63 (1970).
Kristensson, K., Lycke, E., Sjöstrand, J.: Spread of herpes simplex virus in peripheral nerves. Acta neuropath. (Berl.) (in press).
Lasek, J.: Bidirectional transport of radioactively labelled axoplasmic components. Nature (Lond.)216, 1212–1214 (1967).
Lennette, E. H., Koprowski, H.: Influence of age on the susceptibility of mice to infection with certain neurotropic viruses. J. Immunol.49, 175–191 (1944).
Livett, B. G., Geffen, L. B., Austin, L.: Proximo-distal transport of14C-noradrenaline and protein in sympathetic nerves. J. Neurochem.15, 931–939 (1968).
Lubińska, L.: Axoplasmic streaming in regenerating and in normal nerve fibres. In: Mechanisms of Neural Regeneration. Progress in brain research, vol. 13, pp. 1–71. Amsterdam: Elsevier 1964.
Nakai, J.: Dissociated dorsal root ganglia in tissue culture. Amer. J. Anat.99, 81–129 (1956).
Ochs, S., Ranish, N.: Characteristics of the fast transport system in mammalian nerve fibres. J. Neurobiol.1, 247–261 (1969).
Olsson, Y.: Studies on vascular permeability in peripheral nerves. Acta neuropath. (Berl.)7, 1–15 (1966).
Pomerat, C. M., Hendelman, W. J., Raiborn, C. W., Massey, J. F.: Dynamic activities of nervous tissue in vitro. In: The Neuron, pp. 119–178. Amsterdam: Elsevier 1967.
Rinder, L.: Artefactitious extravasation of fluorescent indicators in the investigation of vascular permeability in brain and spinal cord. Acta path. microbiol. scand.74, 333–339 (1968).
Rosenbluth, J., Wissig, S. L.: The distribution of exogenous ferritin in toad spinal ganglia and the mechanism of its uptake by neurons. J. Cell Biol.23, 307–325 (1964).
Sjöstrand, J.: Fast and slow components of axoplasmic transport in the hypoglossal and vagus nerves of the rabbit. Brain Res.18, 461–467 (1970).
Steinwall, O., Klatzo, I.: Double tracer methods in studies on blood-brain barrier dysfunction and brain edema. Acta neurol. scand.41, 591–595 (1965).
Watson, W. E.: Centripetal passage of labelled molecules along mammalian motor axons. J. Physiol. (Lond.)196, 122–123P (1968).
Weiss, P. A.: Neuronal dynamics and neuroplasmic (“axonal”) flow. Symp. Int. Soc. Cell Biol.8, 3–34 (1969).
—, Hiscoe, H.: Experiments on the mechanism of nerve growth. J. exp. Zool.107, 315–396 (1948).
Wildy, P.: The progression of herpes simplex virus to the central nervous system of the mouse. J. Hyg. (Lond.)65, 173–192 (1967).
Wright, G. P.: Nerve trunks as pathways in infection. Proc. roy. Soc. Med.46, 319–330 (1953).
Zelená, J.: Bidirectional shift of mitochondria in axons after injury. Symp. Int. Soc. Cell Biol.8, 73–94 (1969).
Author information
Authors and Affiliations
Additional information
This study was supported by a grant, B70-12X-82-06, from the Swedish Medical Research Council. The author wishes to express his gratitude to Assoc. Prof. P. Sourander and Ass. Prof. Y. Olsson for valuable criticism and support of the study.
Rights and permissions
About this article
Cite this article
Kristensson, K. Transport of fluorescent protein tracer in peripheral nerves. Acta Neuropathol 16, 293–300 (1970). https://doi.org/10.1007/BF00686894
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00686894