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The uptake of horseradish peroxidase by cortical synapses in rat brain

An in vivo study

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Summary

Horseradish peroxidase (HRP) was introduced directly into the cerebral cortex of adult rats, which were allowed to survive for 60 min before perfusion fixation. After the tissue had been incubated to demonstrate HRP at the LM and EM levels, blocks of cortical tissue were taken at varying distances from the injection site. These eight blocks of tissue constituted a time sequence for HRP diffusion.

Qualitative examination of the presynaptic terminals showed that the most commonly encountered profiles are the plain synaptic vesicles, many of which accumulate tracer. In some terminals labelled vesicles are “lined-up” in tubular fashion. Other profiles commonly labelled are coated vesicles, tubular and vacuolar cisternae, and plain and coated pinocytotic vesicles.

Quantitative analyses based on the number of terminals containing labelled profiles demonstrate an early rise in the rate of labelling of both plain synaptic vesicles and coated vesicles, after which synaptic vesicle labelling rises slowly towards a plateau. By contrast, there is a late parallel increase in the rate of labelling of coated vesicles and cisternae. A more detailed analysis, based on the actual numbers of labelled and total profiles within each presynaptic terminal, highlight early and late periods of rapid labelling for plain synaptic vesicles, coated vesicles and cisternae. A further aspect of HRP incorporation studied, concerns its uptake into four delineated regions of the presynaptic terminal.

Our data indicate that membrane uptake into the presynaptic terminal is accomplished mainly via coated vesicles, although plain synaptic vesicles may also be involved. Coated vesicles, in turn, appear to give rise directly to plain synaptic vesicles, with some coalescing to produce vacuolar cisternae. The latter are involved in a two-way interchange of membrane with tubular cisternae, plain synaptic vesicles and coated vesicles. An additional source of plain synaptic vesicles are the tubular cisternae. Exocytosis of plain synaptic vesicles constitutes the mechanism by which transmitter is released from the presynaptic terminal.

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References

  • Albrecht, M.H.: Mounting frozen sections with gelatin. Stain Technol. 29, 89–90 (1954)

    Google Scholar 

  • Baker, R.N., Cancilla, P.A., Pollock, P.S., Frommes, S.P.: The movement of exogenous protein in experimental edema: an electron microscopic study after freeze-injury. J. Neuropath. exp. Neurol. 30, 668–679 (1971)

    Google Scholar 

  • Becker, N.H., Hirano, A., Zimmerman, H.M.: Observations of the distribution of exogenous peroxidase in the rat cerebrum. J. Neuropath. exp. Neurol. 27, 439–452 (1968)

    Google Scholar 

  • Bird, M.M.: Microtubules synaptic vesicle associations in cultured rat spinal cord neurons. Cell Tiss. Res. 168, 101–115 (1976)

    Google Scholar 

  • Brightman, M.W.: The distribution within the brain of ferritin injected into cerebrospinal fluid compartments. I. Ependymal distribution. J. Cell Biol. 26, 99–123 (1965a)

    Google Scholar 

  • Brightman, M.W.: The distribution within the brain of ferritin injected into cerebrospinal fluid compartments. II. Parenchymal distribution. Amer. J. Anat. 117, 193–200 (1965b)

    Google Scholar 

  • Brightman, M.W.: The intracerebral movement of proteins injected into blood and cerebrospinal fluid of mice. Prog. Brain. Res. 29, 19–40 (1968)

    Google Scholar 

  • Broadwell, R.D., Brightman, M.W.: Entry of peroxidase into neurons of the central and peripheral nervous systems from extracerebral and cerebral blood. J. comp. Neurol. 166, 257–284 (1976)

    Google Scholar 

  • Ceccarelli, B., Hurlbut, W.P., Mauro, A.: Turnover of transmitter and synaptic vesicles at the frog neuromuscular junction. J. Cell Biol. 57, 499–524 (1973)

    Google Scholar 

  • Cooke, C.T., Cameron, P.U., Jones, D.G.: Stimulation-induced uptake of horseradish peroxidase by rat cortical synapses. Neurosci. Letters 1, 15–18 (1975)

    Google Scholar 

  • Delorme, P., Gayet, J., Grignon, G.: Diffusion of horseradish peroxidase perfused through the lateral ventricle of the chick telencephalon. Cell Tiss. Res. 157, 535–540 (1975)

    Google Scholar 

  • Droz, B., Rambourg, A., Koenig, H.L.: The smooth endoplasmic reticulum: structure and role in the renewal of axonal membrane and synaptic vesicles by fast axonal transport. Brain Res. 93, 1–13 (1975)

    Google Scholar 

  • Gardner, D.E.: Brain oedema: an experimental model. Brit. J. exp. Path. 55, 453–457 (1974)

    Google Scholar 

  • Gray, E.G.: Pre-synaptic microtubules and their associations with synaptic vesicles. Proc. roy. Soc. B 190, 369–372 (1975)

    Google Scholar 

  • Halperin, J.J., LaVail, J.H.: A study of the dynamics of retrograde transport and accumulation of horseradish peroxidase in injured neurons. Brain Res. 100, 253–269 (1975)

    Google Scholar 

  • Heuser, J.E., Reese, T.S.: Evidence for recycling of synaptic vesicle membrane during transmitter release at the frog neuromuscular junction. J. Cell Biol. 57, 315–344 (1973)

    Google Scholar 

  • Jones, D.G.: Morphological features of central synapses, with emphasis on the presynaptic terminal. Life Sci. (in press, 1977)

  • Jones, D.G., Bradford, H.F.: Observations on the morphology of mammalian synaptosomes following their incubation and electrical stimulation. Brain Res. 28, 491–499 (1971)

    Google Scholar 

  • Jones, D.G., Devon, R.M.: Synaptic ultrastructure under varying degrees of Pentobarbitone anaesthesia. In preparation (1977)

  • Jorgensen, O.S., Mellerup, E.T.: Endocytotic formation of rat brain synaptic vesicles. Nature (Lond.) 249, 770–771 (1974)

    Google Scholar 

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

    Google Scholar 

  • LaVail, J.H.: Retrograde cell degeneration and retrograde transport techniques. In: The use of axonal transport for studies of neuronal connectivity (W.M. Cowan, M. Cuénod, eds.), pp. 217–248. Amsterdam: Elsevier 1975

    Google Scholar 

  • LaVail, J.H., LaVail, M.M.: The retrograde intraaxonal transport of horseradish peroxidase in the chick visual system: a light and electron microscopic study. J. comp. Neurol. 157, 303–358 (1974)

    Google Scholar 

  • Lynch, G., Gall, C., Mensah, P., Cotman, C.W.: Horseradish peroxidase histochemistry: a new method for tracing efferent projections in the central nervous system. Brain Res. 65, 373–380 (1974)

    Google Scholar 

  • Lynch, G., Smith, R.L., Browning, M.D., Deadwyler, S.: Evidence for bidirectional dendritic transport of horseradish peroxidase. Adv. Neurol. 12, 297–313 (1975)

    Google Scholar 

  • Model, P.G., Highstein, S.M., Bennett, M.V.L.: Depletion of vesicles and fatigue of transmission at a vertebrate central synapse. Brain Res. 98, 209–228 (1975)

    Google Scholar 

  • Nauta, H.J.W., Kaiserman-Abramof, I.R., Lasek, R.J.: Electron microscopic observations of horseradish peroxidase transported from the caudoputamen to the substantia nigra in the rat: possible involvement of the agranular reticulum. Brain Res. 85, 373–384 (1975)

    Google Scholar 

  • Nauta, H.J.W., Pritz, M.B., Lasek, R.J.: Afférents to the rat caudoputamen studied with horseradish peroxidase. An evaluation of a retrograde neuroanatomical research method. Brain Res. 67, 219–238 (1974)

    Google Scholar 

  • Nevis, A.H., Collins, G.H.: Electrical impedance and volume changes in brain during preparation for electron microscopy. Brain Res. 5, 57–85 (1967)

    Google Scholar 

  • Ogata, J., Hochwald, G.M., Cravioto, H., Ransohoff, J.: Distribution of intraventricular HRP in normal and hydrocephalic cat brains. J. Neuropath. exp. Neurol. 31, 454–463 (1972)

    Google Scholar 

  • Oppenheim, R.W., Heaton, M.B.: The retrograde transport of horseradish peroxidase from the developing limb of the chick embryo. Brain Res. 98, 291–302 (1975)

    Google Scholar 

  • Pelletier, G., Dupont, A., Puviani, R.: Ultrastructural study of the uptake of peroxidase by the rat median eminence. Cell Tiss. Res. 156, 521–532 (1975)

    Google Scholar 

  • Peters, A.: The fixation of central nervous tissue and the analysis of electron micrographs of the neuropil, with special reference to the cerebral cortex. In: Contemporary research methods in neuroanatomy (W.T.H. Nauta and S.O.E. Ebbeson, eds.), pp. 56–76. Berlin-Heidelberg-New York: Springer 1970

    Google Scholar 

  • Poisner, A.M., Cooke, P.: Microtubules and the adrenal medulla. Ann. N.Y. Acad. Sci. 253, 653–669 (1975)

    Google Scholar 

  • Pysh, J.J., Wiley, R.G.: Synaptic vesicle depletion and recovery in cat sympathetic ganglia electrically stimulated in vivo. Evidence for transmitter secretion by exocytosis. J. Cell Biol. 60, 365–374 (1974)

    Google Scholar 

  • Ralston, H.J., Sharp, P.V.: The identification of thalamocortical relay cells in the adult cat by means of retrograde axonal transport of horseradish peroxidase. Brain Res. 62, 273–278 (1973)

    Google Scholar 

  • Reperant, J.: The orthograde transport of horseradish peroxidase in the visual system. Brain Res. 85, 307–312 (1975)

    Google Scholar 

  • Ripps, H., Shakib, M., MacDonald, E.D.: Peroxidase uptake by photoreceptor terminals of the skate retina. J. Cell Biol. 70, 86–96 (1976)

    Google Scholar 

  • Schacher, S., Holtzman, E., Hood, D.C.: Synaptic activity of frog retinal photoreceptors. A peroxidase uptake study. J. Cell Biol. 70, 178–192 (1976)

    Google Scholar 

  • Selwood, L.: Electron microscopy of the fate of exogenous ferritin in the feline visual cortex. Z. Zellforsch. 107, 6–14 (1970)

    Google Scholar 

  • Sherlock, D.A., Field, P.M., Raisman, G.: Retrograde transport of horseradish peroxidase in the magnocellular neurosecretory system of the rat. Brain Res. 88, 403–414 (1975)

    Google Scholar 

  • Smith, D.S.: On the significance of cross-bridges between microtubules and synaptic vesicles. Phil. Trans. B 261, 395–405 (1971)

    Google Scholar 

  • Stoeckart, R., Jansen, H.G., Kreike, A.J.: Micro-pinocytosis and exocytosis in nerve terminals in the median eminence of the rat. Prog. Brain Res. 41, 111–115 (1974)

    Google Scholar 

  • Turner, P.T., Harris, A.B.: Ultrastructure of synaptic vesicle formation in cerebral cortex. Nature (Lond.) 242, 57–59 (1973)

    Google Scholar 

  • Turner, P.T., Harris, A.B.: Ultrastructure of exogenous peroxidase in cerebral cortex. Brain Res. 74, 305–326 (1974)

    Google Scholar 

  • Whittaker, V.P.: The vesicle hypothesis. In: Excitatory synaptic mechanisms (P. Andersen, J.K.S. Jansen, eds.), pp. 67–76. Oslo: Universitatsforlaget 1970

    Google Scholar 

  • Winfield, D.A., Gatter, K.C., Powell, T.P.S.: An electron microscopic study of retrograde and orthograde transport of horseradish peroxidase to the lateral geniculate nucleus of the monkey. Brain Res. 92, 462–467 (1975)

    Google Scholar 

  • Wooten, G.F., Kopin, I.J., Axelrod, J.: Effects of colchicine and vinblastine on axonal transport and transmitter release in sympathetic nerves. Ann. N.Y. Acad. Sci. 253, 528–533 (1975)

    Google Scholar 

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Supported by the Nuffield Foundation. We are grateful to Mr. M. Austin for help with the photography

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Jones, D.G., Cameron, P.U. & Ellison, L.T. The uptake of horseradish peroxidase by cortical synapses in rat brain. Cell Tissue Res. 178, 355–373 (1977). https://doi.org/10.1007/BF00218700

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