Abstract
The peripheral feeding network of the giant freshwater ameba Reticulomyxa can be easily and rapidly lysed to produce an extensive, stable, and completely exposed cytoskeletal framework of colinear microtubules and microfilaments. Most of the organelles that remain attached to this framework resume rapid saltatory movements at rates of up to 20 micron/s if ATP is added. This lysed model system is also capable of other forms of motility, namely an active splaying of microtubule bundles and bulk streaming. Reactivation does not occur with other nucleoside triphosphates, requires Mg ions, is insensitive to even high concentrations of erythro-9-(3-[2-hydroxynonyl]) adenine, is sensitive to vanadate only at concentrations of approximately 100 microM, and is inhibited by N-ethylmaleimide at concentrations greater than 100 microM. The physiology of this reactivation suggests an organelle transport motor distinct from cytoplasmic dynein and possibly the recently described kinesin. This system can serve as a model for elucidating the mechanisms of intracellular transport and, in addition, provides a unique opportunity to examine associations between microtubules and microfilaments.
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Selected References
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- Allen R. D., Allen N. S., Travis J. L. Video-enhanced contrast, differential interference contrast (AVEC-DIC) microscopy: a new method capable of analyzing microtubule-related motility in the reticulopodial network of Allogromia laticollaris. Cell Motil. 1981;1(3):291–302. doi: 10.1002/cm.970010303. [DOI] [PubMed] [Google Scholar]
- Allen R. D. New observations on cell architecture and dynamics by video-enhanced contrast optical microscopy. Annu Rev Biophys Biophys Chem. 1985;14:265–290. doi: 10.1146/annurev.bb.14.060185.001405. [DOI] [PubMed] [Google Scholar]
- 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]
- Bouchard P., Penningroth S. M., Cheung A., Gagnon C., Bardin C. W. erythro-9-[3-(2-Hydroxynonyl)]adenine is an inhibitor of sperm motility that blocks dynein ATPase and protein carboxylmethylase activities. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1033–1036. doi: 10.1073/pnas.78.2.1033. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brady S. T. A novel brain ATPase with properties expected for the fast axonal transport motor. Nature. 1985 Sep 5;317(6032):73–75. doi: 10.1038/317073a0. [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]
- Cande W. Z., Tooth P. J., Kendrick-Jones J. Regulation of contraction and thick filament assembly-disassembly in glycerinated vertebrate smooth muscle cells. J Cell Biol. 1983 Oct;97(4):1062–1071. doi: 10.1083/jcb.97.4.1062. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clark T. G., Rosenbaum J. L. Pigment particle translocation in detergent-permeabilized melanophores of Fundulus heteroclitus. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4655–4659. doi: 10.1073/pnas.79.15.4655. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Forman D. S. Vanadate inhibits saltatory organelle movement in a permeabilized cell model. Exp Cell Res. 1982 Sep;141(1):139–147. doi: 10.1016/0014-4827(82)90076-3. [DOI] [PubMed] [Google Scholar]
- Gibbons I. R., Cosson M. P., Evans J. A., Gibbons B. H., Houck B., Martinson K. H., Sale W. S., Tang W. J. Potent inhibition of dynein adenosinetriphosphatase and of the motility of cilia and sperm flagella by vanadate. Proc Natl Acad Sci U S A. 1978 May;75(5):2220–2224. doi: 10.1073/pnas.75.5.2220. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gilbert S. P., Allen R. D., Sloboda R. D. Translocation of vesicles from squid axoplasm on flagellar microtubules. Nature. 1985 May 16;315(6016):245–248. doi: 10.1038/315245a0. [DOI] [PubMed] [Google Scholar]
- Inoué S. Video image processing greatly enhances contrast, quality, and speed in polarization-based microscopy. J Cell Biol. 1981 May;89(2):346–356. doi: 10.1083/jcb.89.2.346. [DOI] [PMC free article] [PubMed] [Google Scholar]
- KANE R. E. THE MITOTIC APPARATUS. PHYSICAL-CHEMICAL FACTORS CONTROLLING STABILITY. J Cell Biol. 1965 Apr;25:SUPPL–SUPPL:144. doi: 10.1083/jcb.25.1.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Koonce M. P., Schliwa M. Bidirectional organelle transport can occur in cell processes that contain single microtubules. J Cell Biol. 1985 Jan;100(1):322–326. doi: 10.1083/jcb.100.1.322. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lasek R. J., Brady S. T. Attachment of transported vesicles to microtubules in axoplasm is facilitated by AMP-PNP. Nature. 1985 Aug 15;316(6029):645–647. doi: 10.1038/316645a0. [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]
- Sale W. S., Gibbons I. R. Study of the mechanism of vanadate inhibition of the dynein cross-bridge cycle in sea urchin sperm flagella. J Cell Biol. 1979 Jul;82(1):291–298. doi: 10.1083/jcb.82.1.291. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schliwa M., Ezzell R. M., Euteneuer U. erythro-9-[3-(2-Hydroxynonyl)]adenine is an effective inhibitor of cell motility and actin assembly. Proc Natl Acad Sci U S A. 1984 Oct;81(19):6044–6048. doi: 10.1073/pnas.81.19.6044. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schliwa M. Mechanisms of intracellular organelle transport. Cell Muscle Motil. 1984;5:1-82,403-6. doi: 10.1007/978-1-4684-4592-3_1. [DOI] [PubMed] [Google Scholar]
- Schliwa M., van Blerkom J. Structural interaction of cytoskeletal components. J Cell Biol. 1981 Jul;90(1):222–235. doi: 10.1083/jcb.90.1.222. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scholey J. M., Porter M. E., Grissom P. M., McIntosh J. R. Identification of kinesin in sea urchin eggs, and evidence for its localization in the mitotic spindle. Nature. 1985 Dec 5;318(6045):483–486. doi: 10.1038/318483a0. [DOI] [PubMed] [Google Scholar]
- Sheetz M. P., Spudich J. A. Movement of myosin-coated fluorescent beads on actin cables in vitro. Nature. 1983 May 5;303(5912):31–35. doi: 10.1038/303031a0. [DOI] [PubMed] [Google Scholar]
- Shimizu T., Johnson K. A. Presteady state kinetic analysis of vanadate-induced inhibition of the dynein ATPase. J Biol Chem. 1983 Nov 25;258(22):13833–13840. [PubMed] [Google Scholar]
- Spudich J. A., Kron S. J., Sheetz M. P. Movement of myosin-coated beads on oriented filaments reconstituted from purified actin. Nature. 1985 Jun 13;315(6020):584–586. doi: 10.1038/315584a0. [DOI] [PubMed] [Google Scholar]
- Stearns M. E., Ochs R. L. A functional in vitro model for studies of intracellular motility in digitonin-permeabilized erythrophores. J Cell Biol. 1982 Sep;94(3):727–739. doi: 10.1083/jcb.94.3.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vale R. D., Reese T. S., Sheetz M. P. Identification of a novel force-generating protein, kinesin, involved in microtubule-based motility. Cell. 1985 Aug;42(1):39–50. doi: 10.1016/s0092-8674(85)80099-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vale R. D., Schnapp B. J., Mitchison T., Steuer E., Reese T. S., Sheetz M. P. Different axoplasmic proteins generate movement in opposite directions along microtubules in vitro. Cell. 1985 Dec;43(3 Pt 2):623–632. doi: 10.1016/0092-8674(85)90234-x. [DOI] [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]
- Vale R. D., Schnapp B. J., Reese T. S., Sheetz M. P. Organelle, bead, and microtubule translocations promoted by soluble factors from the squid giant axon. Cell. 1985 Mar;40(3):559–569. doi: 10.1016/0092-8674(85)90204-1. [DOI] [PubMed] [Google Scholar]
- Walter R. J., Berns M. W. Computer-enhanced video microscopy: digitally processed microscope images can be produced in real time. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6927–6931. doi: 10.1073/pnas.78.11.6927. [DOI] [PMC free article] [PubMed] [Google Scholar]