Summary
The dependence of anterograde axoplasmic transport on cytoskeletal components was investigated using microinjection of horseradish peroxidase (HRP) into the somata of chick dorsal root ganglion cells in vitro. Microinjected HRP was transported anterogradely in the neurites and their branches; this transport was disturbed by colchicine in a drug-dependent and time-dependent manner. Cytochalasin B, a drug that depolymerizes actin, did not inhibit the transport of HRP, despite the formation of local swellings in neurites. The microinjection of polyclonal antibodies directed against tubulin and monoclonal antibodies (mAbs) against 200-kDa neurofilaments disturbed the axoplasmic transport of co-injected HRP, which then exhibited an irregular and discontinuous distribution in the axonal branches. The transport of HRP became discontinuous after the injection of anti-tubulin antibodies and led to the formation of globular deposits of HRP. Polyclonal antibodies against actin and mAbs to 160-kDa and 68-kDa neurofilaments seemed to have no effect on the axoplasmic transport of co-injected HRP. Microinjection of antibodies against tubulin induced formation of perinuclear bundles consisting of cytoskeletal components. The transport of HRP thus appears to be regulated by an intact microtubular system and cross-linker components (200-kDa neurofilaments) of the cytoskeleton. Actin and most intermediate filament proteins do not seem to play an essential role in the transport of HRP.
Similar content being viewed by others
References
Adams RJ, Bray D (1983) Rapid transport of foreign particles microinjected into crab axons. Nature 303:718–720
Allen RD, Weiss DG, Hayden JH, Brown DT, Fujiwake H, Simpson M (1985) Gliding movement of an bidirectional transport along single native microtubules from squid axoplasm: evidence for an active role of microtubules in cytoplasmic transport. J Cell Biol 100:1736–1752
Ambron RT, Goldman JE, Schwartz JH (1974) Axonal transport of newly synthesized glyeoproteins in a single identified neuron of Aplysia californica. J Cell Biol 61:665–675
Anderson LE, McClure WO (1973) Differential transport of protein in axons: Comparison between the sciatic nerve and dorsal columns of cats. Proc Natl Acad Sci USA 70:1521–1525
Arnheiter H, Dubois-Dalcq M, Lazzarini RA (1984) Direct visualization of protein transport and processing in the living cell by microinjection of specific antibodies. Cell 39:99–109
Benavente R, Rose KM, Reimer G, Hügle-Dörr B, Scheer U (1987) Inhibition of nucleolar reformation after microinjection of antibodies to RNA polymerase I into mitotic cells. J Cell Biol 105:1483–1491
Berry RW, Schwartz AW (1977) Axonal transport and axonal processing of low molecular weight proteins from the abdominal ganglion of Aplysia. Brain Res 129:75–90
Blose SH, Meltzer DI, Feramisco JR (1984) 10-nm filaments are induced to collapse in living cells microinjected with monoclonal and polyclonal antibodies against tubulin. J Cell Biol 98:847–858
Brady ST, Lasek RJ, Allen RD, Yin HL, Stossel TP (1984) Gelsolin inhibition of fast axonal transport indicates a requirement for actin microfilaments. Nature 310:56–58
Broadwell RD, Oliver C, Brightman MW (1980) Neuronal transport of acid hydrolases and peroxidase within the lysosomal system of organelles: involvement of agranular reticulum-like cisterns. J Comp Neurol 190:519–532
Forman DS (1987) Fast axonal transport: recent developments. Prog Brain Res 71:103–112
Füchtbauer A, Herrmann M, Mandelkow E-M, Jockusch B (1985) Disruption of microtubules in living cells and cell models by high affinity antibodies to beta-tubulin. EMBO J 4:2807–2814
Goldberg DJ (1982) Microinjection into an identified axon to study the mechanism of fast axonal transport. Proc Natl Acad Sci USA 79:4818–4822
Graham RC, Karnovsky MJ (1966) The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney. Ultrastructural correlates by a new technique. J Histochem Cytochem 14:291–302
Graessmann A, Graessmann M (1986) Microinjection of tissue culture cells using glass capillaries: methods. In: Celis JE (ed) Microinjection and organelle transplantation techniques. Methods and applications. Academic Press, London, pp 3–13
Hendrickson AE (1972) Electron microscopic distribution of axoplasmic transport. J Comp Neurol 144:381–397
Hertzberg EL, Spray DC, Bennett MVL (1985) Reduction of gap junctional conductance by microinjection of antibodies against the 27-kDa liver gap junction polypeptide. Proc Natl Acad Sci USA 82:2412–2416
Hirokawa N (1982) Cross-linker system between neurofilaments, microtubules, and membranous organelles in frog axons revealed by the quick-freeze, deep-etching method. J Cell Biol 94:129–142
Isenberg G, Schubert P, Kreutzberg GW (1982) Actin, a neuroplasmic constituent requisite for axonal transport. In: Weiss DG (ed) Axoplasmic transport. Springer, Berlin Heidelberg New York, pp 314–321
Jasmin BJ, Lavoie PA, Gardiner PF (1988) Fast axonal transport of labeled proteins in motoneurons of exercise-trained rats. Am J Physiol 255:731–736
Kenigsberg RL, Trifaró JM (1985) Microinjection of calmodulin antibodies into cultured chromaffin cells blocks catecholamine release in response to stimulation. Neuroscience 14:335–347
Koike H (1987) The disturbance of the fast axonal transport of protein by passive stretching of an axon in Aplysia. J Physiol (Lond) 390:489–500
Koike H, Eisenstadt M, Schwartz JH (1972) Axonal transport of newly synthesized acetylcholine in an identified neuron of Aplysia. Brain Res 37:152–159
Komiya Y, Kurokawa M (1978) Asymmetry of protein transport in two branches of bifurcating axons. Brain Res 139:354–358
Lasek RJ, Brady ST (1982) The structural hypothesis of axonal transport: two classes of moving elements. In: Weiss DG (ed) Axoplasmic transport. Springer, Berlin Heidelberg New York, pp 397–405
McClure WO (1972) Effect of drugs upon axoplasmic transport. Adv Pharmacol Chemotherapy 10:185–220
McEwen BS, Grafstein B (1968) Fast and slow components in axonal transport of protein. J Cell Biol 38:494–508
Meller K (1974) The reaggregation of neurons and their satellite cells in cultures of trypsin-dissociated spinal ganglia. Cell Tissue Res 152:175–183
Mori H, Komiya Y, Kurokawa M (1979) Slowly migrating axonal polypeptides. Inequalities in their rate and amount of transport between two branches of bifurcating axons. J Cell Biol 82:174–184
Morré DJ (1982) Intracellular vesicular transport: Vehicles, guide elements, and mechanisms. In: Weiss DG (ed) Axoplasmic Transport. Springer Berlin Heidelberg New York, pp 2–14
Nemhauser I, Goldberg DJ (1985) Structural effects in axoplasm of DNase I, an actin depolymerizer that blocks fast axonal transport. Brain Res 334:47–58
Ochs S (1972) Rate of fast axoplasmic transport in mammalian nerve fibres. J Physiol (Lond) 227:627–645
Ochs S (1975) Retention and redistribution of proteins in mammalian nerve fibres by axoplasmic transport. J Physiol (Lond) 253:459–475
Ochs S, Erdman J, Jersild RA, McAdoo V (1978) Routing of transported materials in the dorsal root and nerve fiber branches of the dorsal root ganglion. J Neurobiol 9:465–481
Papasozomenos SC, Yoon M, Crane R, Autilio-Gambetti L, Gambetti P (1982) Redistribution of proteins of fast axonal transport following administration of beta, beta'-iminodipropionitrile: a quantitative autoradiographic study. J Cell Biol 95:672–675
Paulson JC, McClure WO (1975) Microtubules and axoplasmic transport. Inhibition of transport by podophyllotoxin: an interaction with microtubule protein. J Cell Biol 67:461–467
Pilgrim C, Stumpf WE (1987) Applications of autoradiography in neurobiological research. J Histochem Cytochem 35:917–928
Price RL, Lasek RJ, Katz MJ (1991) Microtubules have special physical associations with smooth endoplasmic reticula and mitochondria in axons. Brain Res 540:209–216
Schmitt FO (1968) Fibrous proteins — neuronal organelles. Proc Natl Acad Sci USA 60:1092–1101
Schwartz JH, Goldman JE, Ambron RT, Goldberg DJ (1976) Axonal transport of vesicles carrying (3H)serotonin in the metacerebral neuron of Aplysia californica. Cold Spring Harbor Symp Quant Biol 40:83–92
Schwartz JH, Goldberg DJ (1982) Studies on the mechanism of fast axoplasmic transport in single identified neurons. In: Weiss DG (ed) Axoplasmic transport. Springer, Berlin Heidelberg New York, pp 351–361
Seiler M, Weiss DG (1987) Nocodazole irreversibly reduces the capacity of rapid axoplasmic transport in vitro. J Pharmacol Exp Ther 242:277–283
Sjöstrand J (1969) Rapid axoplasmic transport of labelled proteins in the vagus and hypoglossal nerves of the rabbit. Exp Brain Res 8:105–112
Sjöstrand J, Frizell M, Hasselgren P-O (1970) Effects of colchicine on axonal transport in peripheral nerves. J Neurochem 17:1563–1570
Sternberger LA (1986) The unlabeled antibody peroxidase-antiperoxidase (PAP) method. In: Sternberger LA. Immunocytochemistry. Wiley, New York Brisbane Toronto, pp 90–202
Stone GC, Hammerschlag R, Bobinski JA (1984) Involvement of coated vesicles in the initiation of fast axonal transport. Brain Res 291:219–228
Toelle H-G, Weber K, Osborn M (1986) Microinjection of monoclonal antibodies to vimentin, desmin, and GFA in cells which contain more than one IF type. Exp Cell Res 162:462–474
Wehland J, Schröder HC, Weber K (1986) Contribution of microtubules to cellular physiology: microinjection of well-characterized monoclonal antibodies into cultured cells. Ann NY Acad Sci 466:609–621
Wujek JR, Lasek RJ (1983) Correlation of axonal regeneration and slow component B in two branches of a single axon. J Neurosci 3:243–251
Zenker W, Mayr R, Gruber H (1973) Axoplasmic organelles: Quantitative differences between ventral and dorsal root fibres of the rat. Experientia 29:77–78
Zenker W, Mayr R, Gruber H (1975) Neurotubules: Different densities in peripheral motor and sensory nerve fibres. Experientia 31:318–320
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Meller, K. Axoplasmic transport of horseradish peroxidase in single neurons of the dorsal root ganglion studied in vitro by microinjection. Cell Tissue Res. 270, 139–148 (1992). https://doi.org/10.1007/BF00381888
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00381888