Abstract
The distribution and length of actin microfilaments (MF) was determined in axoplasm extruded from the giant axons of the squid (Loligo pealeii). Extruded axoplasm that was separated from the axonal cortex contains approximately 92% of the total axonal actin, and 60% of this actin is polymerized (Morris, J., and R. Lasek. 1984. J. Cell Biol. 98:2064-2076). Localization of MF with rhodamine-phalloidin indicated that the MF were organized in fine columns oriented longitudinally within the axoplasm. In the electron microscope, MF were surrounded by a dense matrix and they were associated with the microtubule domains of the axoplasm. The surrounding matrix tended to obscure the MF which may explain why MF have rarely been recognized before in the inner regions of the axon. The axoplasmic MF are relatively short (number average length of 0.55 micron). Length measurements of MF prepared either in the presence or absence of the actin-filament stabilizing drug phalloidin indicate that axoplasm contains two populations of MF: stable MF (number average length of 0.79 micron) and metastable MF (number average length of 0.41 micron). Although individual axonal MF are much shorter than axonal microtubules, the combined length of the total MF is twice that of the total microtubules. Apparently, these numerous short MF have an important structural role in the architecture of the inner axonal cytoskeleton.
Full Text
The Full Text of this article is available as a PDF (2.4 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Allen R. D., Weiss D. G., Hayden J. H., Brown D. T., Fujiwake H., Simpson M. Gliding movement of and bidirectional transport along single native microtubules from squid axoplasm: evidence for an active role of microtubules in cytoplasmic transport. J Cell Biol. 1985 May;100(5):1736–1752. doi: 10.1083/jcb.100.5.1736. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Alonso G., Gabrion J., Travers E., Assenmacher I. Ultrastructural organization of actin filaments in neurosecretory axons of the rat. Cell Tissue Res. 1981;214(2):323–341. doi: 10.1007/BF00249215. [DOI] [PubMed] [Google Scholar]
- Brady S. T., Lasek R. J., Allen R. D., Yin H. L., Stossel T. P. Gelsolin inhibition of fast axonal transport indicates a requirement for actin microfilaments. Nature. 1984 Jul 5;310(5972):56–58. doi: 10.1038/310056a0. [DOI] [PubMed] [Google Scholar]
- Brady S. T., Lasek R. J. Nerve-specific enolase and creatine phosphokinase in axonal transport: soluble proteins and the axoplasmic matrix. Cell. 1981 Feb;23(2):515–523. doi: 10.1016/0092-8674(81)90147-1. [DOI] [PubMed] [Google Scholar]
- Bray D., Bunge M. B. Serial analysis of microtubules in cultured rat sensory axons. J Neurocytol. 1981 Aug;10(4):589–605. doi: 10.1007/BF01262592. [DOI] [PubMed] [Google Scholar]
- Clarke F. M., Masters C. J. On the association of glycolytic enzymes with structural proteins of skeletal muscle. Biochim Biophys Acta. 1975 Jan 13;381(1):37–46. doi: 10.1016/0304-4165(75)90187-7. [DOI] [PubMed] [Google Scholar]
- Cooper J. A., Pollard T. D. Methods to measure actin polymerization. Methods Enzymol. 1982;85(Pt B):182–210. doi: 10.1016/0076-6879(82)85021-0. [DOI] [PubMed] [Google Scholar]
- Dancker P., Löw I., Hasselbach W., Wieland T. Interaction of actin with phalloidin: polymerization and stabilization of F-actin. Biochim Biophys Acta. 1975 Aug 19;400(2):407–414. doi: 10.1016/0005-2795(75)90196-8. [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]
- Heriot K., Gambetti P., Lasek R. J. Proteins transported in slow components a and b of axonal transport are distributed differently in the transverse plane of the axon. J Cell Biol. 1985 Apr;100(4):1167–1172. doi: 10.1083/jcb.100.4.1167. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hirokawa N. Cross-linker system between neurofilaments, microtubules, and membranous organelles in frog axons revealed by the quick-freeze, deep-etching method. J Cell Biol. 1982 Jul;94(1):129–142. doi: 10.1083/jcb.94.1.129. [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]
- Kawamura M., Maruyama K. Electron microscopic particle length of F-actin polymerized in vitro. J Biochem. 1970 Mar;67(3):437–457. doi: 10.1093/oxfordjournals.jbchem.a129267. [DOI] [PubMed] [Google Scholar]
- Knull H. R., Bronstein W. W., DesJardins P., Niehaus W. G., Jr Interaction of selected brain glycolytic enzymes with an F-actin-tropomyosin complex. J Neurochem. 1980 Jan;34(1):222–225. doi: 10.1111/j.1471-4159.1980.tb04646.x. [DOI] [PubMed] [Google Scholar]
- Korn E. D. Actin polymerization and its regulation by proteins from nonmuscle cells. Physiol Rev. 1982 Apr;62(2):672–737. doi: 10.1152/physrev.1982.62.2.672. [DOI] [PubMed] [Google Scholar]
- LeBeux Y. J., Willemot J. An ultrastructural study of the microfilaments in rat brain by means of heavy meromyosin labeling. I. The perikaryon, the dendrites and the axon. Cell Tissue Res. 1975 Jun 27;160(1):1–36. doi: 10.1007/BF00219840. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Letourneau P. C. Differences in the organization of actin in the growth cones compared with the neurites of cultured neurons from chick embryos. J Cell Biol. 1983 Oct;97(4):963–973. doi: 10.1083/jcb.97.4.963. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maupin P., Pollard T. D. Arrangement of actin filaments and myosin-like filaments in the contractile ring and of actin-like filaments in the mitotic spindle of dividing HeLa cells. J Ultrastruct Mol Struct Res. 1986 Jan;94(1):92–103. doi: 10.1016/0889-1605(86)90055-8. [DOI] [PubMed] [Google Scholar]
- McDonald K. Osmium ferricyanide fixation improves microfilament preservation and membrane visualization in a variety of animal cell types. J Ultrastruct Res. 1984 Feb;86(2):107–118. doi: 10.1016/s0022-5320(84)80051-9. [DOI] [PubMed] [Google Scholar]
- McQuarrie I. G., Brady S. T., Lasek R. J. Diversity in the axonal transport of structural proteins: major differences between optic and spinal axons in the rat. J Neurosci. 1986 Jun;6(6):1593–1605. doi: 10.1523/JNEUROSCI.06-06-01593.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Metuzals J., Hodge A. J., Lasek R. J., Kaiserman-Abramof I. R. Neurofilamentous network and filamentous matrix preserved and isolated by different techniques from squid giant axon. Cell Tissue Res. 1983;228(3):415–432. doi: 10.1007/BF00211465. [DOI] [PubMed] [Google Scholar]
- Metuzals J., Tasaki I. Subaxolemmal filamentous network in the giant nerve fiber of the squid (Loligo pealei L.) and its possible role in excitability. J Cell Biol. 1978 Aug;78(2):597–621. doi: 10.1083/jcb.78.2.597. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morris J. R., Lasek R. J. Monomer-polymer equilibria in the axon: direct measurement of tubulin and actin as polymer and monomer in axoplasm. J Cell Biol. 1984 Jun;98(6):2064–2076. doi: 10.1083/jcb.98.6.2064. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morris J. R., Lasek R. J. Stable polymers of the axonal cytoskeleton: the axoplasmic ghost. J Cell Biol. 1982 Jan;92(1):192–198. doi: 10.1083/jcb.92.1.192. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nemhauser I., Goldberg D. J. Structural effects in axoplasm of DNase I, an actin depolymerizer that blocks fast axonal transport. Brain Res. 1985 May 13;334(1):47–58. doi: 10.1016/0006-8993(85)90566-9. [DOI] [PubMed] [Google Scholar]
- Papasozomenos S. C., Payne M. R. Actin immunoreactivity localizes with segregated microtubules and membraneous organelles and in the subaxolemmal region in the beta,beta'-iminodipropionitrile axon. J Neurosci. 1986 Dec;6(12):3483–3491. doi: 10.1523/JNEUROSCI.06-12-03483.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sattilaro R. F., Dentler W. L., LeCluyse E. L. Microtubule-associated proteins (MAPs) and the organization of actin filaments in vitro. J Cell Biol. 1981 Aug;90(2):467–473. doi: 10.1083/jcb.90.2.467. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scordilis S. P., Adelstein R. S. A comparative study of the myosin light chain kinases from myoblast and muscle sources. Studies on the kinases from proliferative rat myoblasts in culture, rat thigh muscle, and rabbit skeletal muscle. J Biol Chem. 1978 Dec 25;253(24):9041–9048. [PubMed] [Google Scholar]
- Shaw G., Osborn M., Weber K. Arrangement of neurofilaments, microtubules and microfilament-associated proteins in cultured dorsal root ganglia cells. Eur J Cell Biol. 1981 Apr;24(1):20–27. [PubMed] [Google Scholar]
- Shen B. W., Josephs R., Steck T. L. Ultrastructure of unit fragments of the skeleton of the human erythrocyte membrane. J Cell Biol. 1984 Sep;99(3):810–821. doi: 10.1083/jcb.99.3.810. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spooner B. S., Holladay C. R. Distribution of tubulin and actin in neurites and growth cones of differentiating nerve cells. Cell Motil. 1981;1(2):167–178. doi: 10.1002/cm.970010202. [DOI] [PubMed] [Google Scholar]
- Tsukita S., Tsukita S., Kobayashi T., Matsumoto G. Subaxolemmal cytoskeleton in squid giant axon. II. Morphological identification of microtubule- and microfilament-associated domains of axolemma. J Cell Biol. 1986 May;102(5):1710–1725. doi: 10.1083/jcb.102.5.1710. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vale R. D., Schnapp B. J., Reese T. S., Sheetz M. P. Movement of organelles along filaments dissociated from the axoplasm of the squid giant axon. Cell. 1985 Feb;40(2):449–454. doi: 10.1016/0092-8674(85)90159-x. [DOI] [PubMed] [Google Scholar]
- Weeds A. G., Taylor R. S. Separation of subfragment-1 isoenzymes from rabbit skeletal muscle myosin. Nature. 1975 Sep 4;257(5521):54–56. doi: 10.1038/257054a0. [DOI] [PubMed] [Google Scholar]
- Wulf E., Deboben A., Bautz F. A., Faulstich H., Wieland T. Fluorescent phallotoxin, a tool for the visualization of cellular actin. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4498–4502. doi: 10.1073/pnas.76.9.4498. [DOI] [PMC free article] [PubMed] [Google Scholar]