Abstract
Light and high voltage electron microscopy (HVEM) procedures have been employed to examine the processes regulating saltatory motion in neurons. Light microscope studies demonstrate that organelle transport occurs by rapid bidirectional saltations along linear pathways in cultured neuroblastoma cells. HVEM stereo images of axons reveal that microtubules (Mts) and organelles are suspended in a continuous latticework of fine microtrabecular filaments and that the Mts and lattice constitute a basic cytoskeletal structure mediating the motion of particles along axons. We propose that particle transport depends on dynamic properties of nonstatic microtrabecular lattice components. EXperiments were initiated to determine the effects of changes in divalent cation concentrations (Ca2+ and Mg2+) on: (a)the continuation of transport and (b) the corresponding structural properties of the microtrabecular lattice. We discovered that transport continues or is stimulated to a limited extent in cells exposed to small amounts of exogenously supplied Ca2+ and Mg2+ ions (less than 0.1 mM). Exposure of neurons to increased dosages of Ca2+ and Mg2+ (0.2-1.0 mM) stimulates transport for 2-4 min at 37 degrees C, but after a 5- to 20-min exposure the saltatory movements of organelles are observed gradually to become shorter in duration and rate particle motion ceases to occur. HVEM observations demonstrated that Ca2+ - and with the cessation of motion. Ca2+-containing solutions produced contractions of the microtrabecular filaments, whereas Mg2+-containing solutions had the opposing effect of stimulating an elongation and assembly (expansion) of microtrabeculae. On the basis of these observations we hypothesize that cycles of Ca2+/Mg2+-coupled contractions and expansions of the microtrabecular lattice probably regulate organelle motion in nerve cells.
Full Text
The Full Text of this article is available as a PDF (1.7 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Berlinrood M., McGee-Russell S. M., Allen R. D. Patterns of particle movement in nerve fibres in vitro. An analysis by photokymography and microscopy. J Cell Sci. 1972 Nov;11(3):875–886. doi: 10.1242/jcs.11.3.875. [DOI] [PubMed] [Google Scholar]
- Brady S. T., Crothers S. D., Nosal C., McClure W. O. Fast axonal transport in the presence of high Ca2+: evidence that microtubules are not required. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5909–5913. doi: 10.1073/pnas.77.10.5909. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Buckley I. K., Porter K. R. Electron microscopy of critical point dried whole cultured cells. J Microsc. 1975 Jul;104(2):107–120. doi: 10.1111/j.1365-2818.1975.tb04010.x. [DOI] [PubMed] [Google Scholar]
- Buckley I. K. Three dimensional fine structure of cultured cells: possible implications for subcellular motility. Tissue Cell. 1975;7(1):51–72. doi: 10.1016/s0040-8166(75)80007-3. [DOI] [PubMed] [Google Scholar]
- Burton P. R., Fernandez H. L. Delineation by lanthanum staining of filamentous elements associated with the surfaces of axonal microtubules. J Cell Sci. 1973 Mar;12(2):567–583. doi: 10.1242/jcs.12.2.567. [DOI] [PubMed] [Google Scholar]
- Byers H. R., Porter K. R. Transformations in the structure of the cytoplasmic ground substance in erythrophores during pigment aggregation and dispersion. I. A study using whole-cell preparations in stereo high voltage electron microscopy. J Cell Biol. 1977 Nov;75(2 Pt 1):541–558. doi: 10.1083/jcb.75.2.541. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Byers M. R. Structural correlates of rapid axonal transport: evidence that microtubules may not be directly involved. Brain Res. 1974 Jul 19;75(1):97–113. doi: 10.1016/0006-8993(74)90773-2. [DOI] [PubMed] [Google Scholar]
- Chang C. M., Goldman R. D. The localization of actin-like fibers in cultured neuroblastoma cells as revealed by heavy meromyosin binding. J Cell Biol. 1973 Jun;57(3):867–874. doi: 10.1083/jcb.57.3.867. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Douglas W. W., Sorimachi M. Colchicine inhibits adrenal medullary secretion evoked by acetylcholine without affecting that evoked by potassium. Br J Pharmacol. 1972 May;45(1):129–132. [PMC free article] [PubMed] [Google Scholar]
- Ellisman M. H., Porter K. R. Microtrabecular structure of the axoplasmic matrix: visualization of cross-linking structures and their distribution. J Cell Biol. 1980 Nov;87(2 Pt 1):464–479. doi: 10.1083/jcb.87.2.464. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fernandez H. L., Samson F. E. Axoplasmic transport: differential inhibition by cytochalasin-B. J Neurobiol. 1973;4(3):201–206. doi: 10.1002/neu.480040305. [DOI] [PubMed] [Google Scholar]
- Forman D. S., Padjen A. L., Siggins G. R. Axonal transport of organelles visualized by light microscopy: cinemicrographic and computer analysis. Brain Res. 1977 Nov 11;136(2):197–213. doi: 10.1016/0006-8993(77)90798-3. [DOI] [PubMed] [Google Scholar]
- Forman D. S., Padjen A. L., Siggins G. R. Effect of temperature on the rapid retrograde transport of microscopically visible intra-axonal organelles. Brain Res. 1977 Nov 11;136(2):215–226. doi: 10.1016/0006-8993(77)90799-5. [DOI] [PubMed] [Google Scholar]
- Freed J. J., Lebowitz M. M. The association of a class of saltatory movements with microtubules in cultured cells. J Cell Biol. 1970 May;45(2):334–354. doi: 10.1083/jcb.45.2.334. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Furcht L. T., Scott E. Effect of vinblastine sulfate, colchicine and lumicolchicine on membrane organization of normal and transformed cells. Exp Cell Res. 1975 Dec;96(2):271–282. doi: 10.1016/0014-4827(75)90257-8. [DOI] [PubMed] [Google Scholar]
- Gabbay K. H., Tze W. J. Inhibition of glucose-induced release of insulin by aldose reductase inhibitors. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1435–1439. doi: 10.1073/pnas.69.6.1435. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gibbons B. H., Gibbons I. R. The effect of partial extraction of dynein arms on the movement of reactivated sea-urchin sperm. J Cell Sci. 1973 Sep;13(2):337–357. doi: 10.1242/jcs.13.2.337. [DOI] [PubMed] [Google Scholar]
- Goldberg D. J., Harris D. A., Lubit B. W., Schwartz J. H. Analysis of the mechanism of fast axonal transport by intracellular injection of potentially inhibitory macromolecules: evidence for a possible role of actin filaments. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7448–7452. doi: 10.1073/pnas.77.12.7448. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Griffith L. M., Pollard T. D. Evidence for actin filament-microtubule interaction mediated by microtubule-associated proteins. J Cell Biol. 1978 Sep;78(3):958–965. doi: 10.1083/jcb.78.3.958. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hammerschlag R., Dravid A. R., Chiu A. Y. Mechanism of axonal transport: a proposed role for calcium ions. Science. 1975 Apr 18;188(4185):273–275. doi: 10.1126/science.47182. [DOI] [PubMed] [Google Scholar]
- Henkart M. P., Reese T. S., Brinley F. J., Jr Endoplasmic reticulum sequesters calcium in the squid giant axon. Science. 1978 Dec 22;202(4374):1300–1303. doi: 10.1126/science.725607. [DOI] [PubMed] [Google Scholar]
- Heslop J. P. Axonal flow and fast transport in nerves. Adv Comp Physiol Biochem. 1975;6:75–163. doi: 10.1016/b978-0-12-011506-8.50008-1. [DOI] [PubMed] [Google Scholar]
- Hinkley R. E., Jr Axonal microtubules and associated filaments stained by Alcian blue. J Cell Sci. 1973 Nov;13(3):753–761. doi: 10.1242/jcs.13.3.753. [DOI] [PubMed] [Google Scholar]
- Hoffman P. N., Lasek R. J. The slow component of axonal transport. Identification of major structural polypeptides of the axon and their generality among mammalian neurons. J Cell Biol. 1975 Aug;66(2):351–366. doi: 10.1083/jcb.66.2.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Isenberg G., Schubert P., Kreutzberg G. W. Experimental approach to test the role of actin in axonal transport. Brain Res. 1980 Aug 4;194(2):588–593. doi: 10.1016/0006-8993(80)91247-0. [DOI] [PubMed] [Google Scholar]
- Kreutzberg G. W. Neuronal dynamics and axonal flow. IV. Blockage of intra-axonal enzyme transport by colchicine. Proc Natl Acad Sci U S A. 1969 Mar;62(3):722–728. doi: 10.1073/pnas.62.3.722. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kuczmarski E. R., Rosenbaum J. L. Studies on the organization and localization of actin and myosin in neurons. J Cell Biol. 1979 Feb;80(2):356–371. doi: 10.1083/jcb.80.2.356. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lane N. J., Treherne J. E. Lanthanum staining of neurotubules in axons from cockroach ganglia. J Cell Sci. 1970 Jul;7(1):217–231. doi: 10.1242/jcs.7.1.217. [DOI] [PubMed] [Google Scholar]
- Le Breton G. C., Dinerstein R. J., Roth L. J., Feinberg H. Direct evidence for intracellular divalent cation redistribution associated with platelet shape change. Biochem Biophys Res Commun. 1976 Jul 12;71(1):362–370. doi: 10.1016/0006-291x(76)90291-6. [DOI] [PubMed] [Google Scholar]
- Lubińska L. On axoplasmic flow. Int Rev Neurobiol. 1975;17:241–296. doi: 10.1016/s0074-7742(08)60211-1. [DOI] [PubMed] [Google Scholar]
- Luby K. J., Porter K. R. The control of pigment migration in isolated erythrophores of Holocentrus ascensionis (Osbeck). I. Energy requirements. Cell. 1980 Aug;21(1):13–23. doi: 10.1016/0092-8674(80)90110-5. [DOI] [PubMed] [Google Scholar]
- Metuzals J., Izzard C. S. Spatial patterns of threadlike elements in the axoplasm of the giant nerve fiber of the squid (Loligo pealii L.) as disclosed by differential interference microscopy and by electron microscopy. J Cell Biol. 1969 Dec;43(3):456–479. doi: 10.1083/jcb.43.3.456. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moran D., Rice R. W. Action of papaverine and ionophore A23187 on neurulation. Nature. 1976 Jun 10;261(5560):497–499. doi: 10.1038/261497a0. [DOI] [PubMed] [Google Scholar]
- Ochs S., Hollingsworth D. Dependence of fast axoplasmic transport in nerve on oxidative metabolism. J Neurochem. 1971 Jan;18(1):107–114. doi: 10.1111/j.1471-4159.1971.tb00172.x. [DOI] [PubMed] [Google Scholar]
- Samson F. E., Jr Mechanism of axoplasmic transport. J Neurobiol. 1971;2(4):347–360. doi: 10.1002/neu.480020407. [DOI] [PubMed] [Google Scholar]
- Schmitt F. O. Fibrous proteins--neuronal organelles. Proc Natl Acad Sci U S A. 1968 Aug;60(4):1092–1101. doi: 10.1073/pnas.60.4.1092. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Seeds N. W., Gilman A. G., Amano T., Nirenberg M. W. Regulation of axon formation by clonal lines of a neural tumor. Proc Natl Acad Sci U S A. 1970 May;66(1):160–167. doi: 10.1073/pnas.66.1.160. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shigenaka Y., Watanabe K., Kaneda M. Degrading and stabilizing effects of Mg2+ ions on microtubule-containing axopodia. Exp Cell Res. 1974 Apr;85(2):391–398. doi: 10.1016/0014-4827(74)90141-4. [DOI] [PubMed] [Google Scholar]
- Tani E., Ametani T. Substructure of microtubules in brain nerve cells as revealed by ruthenium red. J Cell Biol. 1970 Jul;46(1):159–165. doi: 10.1083/jcb.46.1.159. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Trifaró J. M., Collier B., Lastowecka A., Stern D. Inhibition by colchicine and by vinblastine of acetylcholine-induced catecholamine release from the adrenal gland: an anticholinergic action, not an effect upon microtubules. Mol Pharmacol. 1972 Mar;8(2):264–267. [PubMed] [Google Scholar]
- Wilson L., Bryan J., Ruby A., Mazia D. Precipitation of proteins by vinblastine and calcium ions. Proc Natl Acad Sci U S A. 1970 Jul;66(3):807–814. doi: 10.1073/pnas.66.3.807. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wolosewick J. J., Porter K. R. Microtrabecular lattice of the cytoplasmic ground substance. Artifact or reality. J Cell Biol. 1979 Jul;82(1):114–139. doi: 10.1083/jcb.82.1.114. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wuerker R. B., Kirkpatrick J. B. Neuronal microtubules, neurofilaments, and microfilaments. Int Rev Cytol. 1972;33:45–75. doi: 10.1016/s0074-7696(08)61448-5. [DOI] [PubMed] [Google Scholar]
- Wunderlich F., Müller R., Speth V. Direct evidence for a colchicine-induced impairment in the mobility of membrane components. Science. 1973 Dec 14;182(4117):1136–1138. doi: 10.1126/science.182.4117.1136. [DOI] [PubMed] [Google Scholar]
- Yamada K. M., Spooner B. S., Wessells N. K. Ultrastructure and function of growth cones and axons of cultured nerve cells. J Cell Biol. 1971 Jun;49(3):614–635. doi: 10.1083/jcb.49.3.614. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zanetti N. C., Mitchell D. R., Warner F. D. Effects of divalent cations on dynein cross bridging and ciliary microtubule sliding. J Cell Biol. 1979 Mar;80(3):573–588. doi: 10.1083/jcb.80.3.573. [DOI] [PMC free article] [PubMed] [Google Scholar]