Home LiteratureArticle Details
PMID: 2670955 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Probing the role of nonmuscle tropomyosin isoforms in intracellular granule movement by microinjection of monoclonal antibodies.

The Journal of cell biology ·Vol. 109 ·No. 3 ·1989-09-00 ·Pages 1141-52

Hegmann TE, Lin JL, Lin JJ

Abstract

Chicken embryo fibroblast (CEF) cells were microinjected with several different monoclonal antibodies that recognize certain nonmuscle isoforms of tropomyosin. Immediately after injection, cells were recorded with a time-lapse video imaging system; later analysis of the tapes revealed that particles in cells injected with one of these antibodies (CG1, specific for CEF tropomyosin isoforms 1 and 3) showed a dramatic decrease in instantaneous speed while moving, distance moved per saltation, and proportion of time spent in motion. Injection of Fab fragments of CG1 resulted in similar changes in the pattern of granule movement. This inhibition of granule movement by CG1 antibody was reversible; at 2.5 h after injection, granules in injected cells had already reached three-fourths of normal speed. The speed of granule movement in cells injected either with antibody specific for tropomyosin isoforms not present in CEF cells, or with CG1 antibody preabsorbed with tropomyosin, was not significantly different from the speed of granules in uninjected cells. When cells were injected with CG1 or Fab fragments of CG1, fixed, and counter-stained with rabbit antibodies to reveal the microtubule, microfilament, and intermediate filament systems, no obvious differences from the patterns normally seen in uninjected cells were observed. Examination of the ultrastructure of injected cells by EM confirmed the presence of apparently intact and normal microtubule, actin, and intermediate filament networks. These experiments suggest that tropomyosin may play an important role in the movement of vesicles and organelles in the cell cytoplasm. Also, we have shown previously that the CG1 determinant can undergo a motility-dependent change in reactivity, that may be important for the regulatory function of nonmuscle tropomyosin (Hegmann, T. E., J. L.-C. Lin, and J. J.-C. Lin. 1988. J. Cell Biol. 106:385-393). Therefore, in addition to postulated microtubule-based motors, microfilaments may play a critical role in regulating granule movement in nonmuscle cells.

MeSH Terms
Actin Cytoskeleton/physiology,ultrastructure Animals Antibodies, Monoclonal Antigen-Antibody Complex Blotting, Western Cells, Cultured Chick Embryo Cytoplasmic Granules/physiology,ultrastructure Fibroblasts/physiology,ultrastructure Fluorescent Antibody Technique Intermediate Filaments/physiology,ultrastructure Microinjections Microscopy, Electron Microtubules/physiology,ultrastructure Molecular Weight Tropomyosin/immunology,isolation & purification,physiology
Chemicals
Antibodies, Monoclonal Antigen-Antibody Complex Tropomyosin
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Hegmann T E
Department of Biology, University of Iowa, Iowa City 52242.
Lin J L
Lin J J
References (53)
53 references, click to expand
  1. Single microtubules from squid axoplasm support bidirectional movement of organelles.
    Cell. 1985 Feb;40(2):455-62 PMID: 2578325
  2. Differential localization of tropomyosin isoforms in cultured nonmuscle cells.
    J Cell Biol. 1988 Aug;107(2):563-72 PMID: 3047141
  3. 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):1736-52 PMID: 2580845
  4. Identification of a novel force-generating protein, kinesin, involved in microtubule-based motility.
    Cell. 1985 Aug;42(1):39-50 PMID: 3926325
  5. Monoclonal antibodies against chicken tropomyosin isoforms: production, characterization, and application.
    Hybridoma. 1985 Fall;4(3):223-42 PMID: 3899907
  6. Identification of a secretory granule-binding protein as caldesmon.
    Nature. 1986 Jan 2-8;319(6048):68-70 PMID: 3941739
  7. Thin filament regulatory proteins of smooth- and non-muscle cells.
    Nature. 1986 Jun 19-25;321(6072):726-7 PMID: 3713859
  8. Identification of a MAP 2-like ATP-binding protein associated with axoplasmic vesicles that translocate on isolated microtubules.
    J Cell Biol. 1986 Sep;103(3):947-56 PMID: 3091608
  9. Stable complexes of axoplasmic vesicles and microtubules: protein composition and ATPase activity.
    J Cell Biol. 1986 Sep;103(3):957-68 PMID: 2943747
  10. Propulsion of organelles isolated from Acanthamoeba along actin filaments by myosin-I.
    Nature. 1986 Aug 21-27;322(6081):754-6 PMID: 3748157
  11. Microtubules and the endoplasmic reticulum are highly interdependent structures.
    J Cell Biol. 1986 Oct;103(4):1557-68 PMID: 3533956
  12. Chromosomes move poleward in anaphase along stationary microtubules that coordinately disassemble from their kinetochore ends.
    J Cell Biol. 1987 Jan;104(1):9-18 PMID: 3793763
  13. The directed migration of eukaryotic cells.
    Annu Rev Cell Biol. 1986;2:337-65 PMID: 3548771
  14. Translocation and clustering of endosomes and lysosomes depends on microtubules.
    J Cell Biol. 1987 Sep;105(3):1253-65 PMID: 3308906
  15. MAP 1C is a microtubule-activated ATPase which translocates microtubules in vitro and has dynein-like properties.
    J Cell Biol. 1987 Sep;105(3):1273-82 PMID: 2958482
  16. The role of microtubule polarity in the movement of kinesin and kinetochores.
    J Cell Sci Suppl. 1986;5:121-8 PMID: 3115997
  17. Motility-dependence of the heterogenous staining of culture cells by a monoclonal anti-tropomyosin antibody.
    J Cell Biol. 1988 Feb;106(2):385-93 PMID: 2448313
  18. The microtubule-dependent formation of a tubulovesicular network with characteristics of the ER from cultured cell extracts.
    Cell. 1988 Jul 1;54(1):27-35 PMID: 3289756
  19. Dynamic behavior of endoplasmic reticulum in living cells.
    Cell. 1988 Jul 1;54(1):37-46 PMID: 3383243
  20. The association of a class of saltatory movements with microtubules in cultured cells.
    J Cell Biol. 1970 May;45(2):334-54 PMID: 5513607
  21. Control of muscle contraction.
    Q Rev Biophys. 1969 Nov;2(4):351-84 PMID: 4935801
  22. Polarized intracellular particle transport: saltatory movements and cytoplasmic streaming.
    Int Rev Cytol. 1972;32:93-137 PMID: 4114619
  23. The role of microfilaments and microtubules in cell movement, endocytosis and exocytosis.
    Ciba Found Symp. 1973;14:109-48 PMID: 4591633
  24. Microtubules in intracellular locomotion.
    Ciba Found Symp. 1973;14:149-69 PMID: 4130533
  25. The role of microtubules in the movement of pigment granules in teleost melanophores.
    J Cell Biol. 1974 Jun;61(3):757-79 PMID: 4836391
  26. Association of actin and myosin with secretory granule membranes.
    Nature. 1975 Apr 10;254(5500):526-9 PMID: 1121327
  27. Regulation of muscular contraction. Distribution of actin control and myosin control in the animal kingdom.
    J Gen Physiol. 1975 Jul;66(1):1-30 PMID: 125778
  28. Endocytosis.
    Annu Rev Biochem. 1977;46:669-722 PMID: 332066
  29. Nonmuscle contractile proteins: the role of actin and myosin in cell motility and shape determination.
    Annu Rev Biochem. 1977;46:797-822 PMID: 143236
  30. Functions of cytoplasmic fibers in intracellular movements in BHK-21 cells.
    J Cell Biol. 1978 Dec;79(3):708-26 PMID: 569659
  31. Microinjection of early SV40 DNA fragments and T antigen.
    Methods Enzymol. 1980;65(1):816-25 PMID: 6246377
  32. Experimental approach to test the role of actin in axonal transport.
    Brain Res. 1980 Aug 4;194(2):588-93 PMID: 6155979
  33. 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):5909-13 PMID: 6160585
  34. 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):7448-52 PMID: 6164061
  35. Intermediate filaments in 3T3 cells collapse after intracellular injection of a monoclonal anti-intermediate filament antibody.
    Nature. 1981 May 21;291(5812):249-51 PMID: 6785655
  36. Disruption of the in vivo distribution of the intermediate filaments in fibroblasts through the microinjection of a specific monoclonal antibody.
    Cell. 1981 Apr;24(1):185-93 PMID: 7016335
  37. Conformational dynamics of tropomyosin in solution: evidence for two conformational states.
    Ann N Y Acad Sci. 1981;366:285-99 PMID: 6942749
  38. Purification of a calmodulin-binding protein from chicken gizzard that interacts with F-actin.
    Proc Natl Acad Sci U S A. 1981 Sep;78(9):5652-5 PMID: 6946503
  39. Coiling of intermediate filaments induced by microinjection of a vimentin-specific antibody does not interfere with locomotion and mitosis.
    Eur J Cell Biol. 1981 Dec;26(1):83-90 PMID: 7198976
  40. Chromaffin granule membrane-F-actin interactions are calcium sensitive.
    Nature. 1982 Jan 28;295(5847):336-9 PMID: 7057898
  41. 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):129-42 PMID: 6181077
  42. Microtubule-organizing centers and cell migration: effect of inhibition of migration and microtubule disruption in endothelial cells.
    J Cell Biol. 1983 May;96(5):1266-72 PMID: 6341378
  43. Movement of myosin-coated fluorescent beads on actin cables in vitro.
    Nature. 1983 May 5-11;303(5912):31-5 PMID: 6682486
  44. Cytoplasmic transport in keratocytes: direct visualization of particle translocation along microtubules.
    Cell Motil. 1983;3(1):1-19 PMID: 6601992
  45. A rat monoclonal antibody reacting specifically with the tyrosylated form of alpha-tubulin. II. Effects on cell movement, organization of microtubules, and intermediate filaments, and arrangement of Golgi elements.
    J Cell Biol. 1983 Nov;97(5 Pt 1):1476-90 PMID: 6685128
  46. A time-lapse video image intensification analysis of cytoplasmic organelle movements during endosome translocation.
    J Cell Biol. 1984 Feb;98(2):565-76 PMID: 6693496
  47. Tropomyosin-enriched and alpha-actinin-enriched microfilaments isolated from chicken embryo fibroblasts by monoclonal antibodies.
    J Cell Biol. 1984 Jan;98(1):116-27 PMID: 6538570
  48. Gelsolin inhibition of fast axonal transport indicates a requirement for actin microfilaments.
    Nature. 1984 Jul 5-11;310(5972):56-8 PMID: 6204208
  49. Persistent, directional motility of cells and cytoplasmic fragments in the absence of microtubules.
    Nature. 1984 Jul 5-11;310(5972):58-61 PMID: 6377086
  50. Chromatophores--models for studying cytomatrix translocations.
    J Cell Biol. 1984 Jul;99(1 Pt 2):152s-158s PMID: 6746727
  51. Interaction of rat liver lysosomal membranes with actin.
    J Cell Biol. 1984 Aug;99(2):680-5 PMID: 6746742
  52. Monoclonal antibodies against caldesmon, a Ca++/calmodulin- and actin-binding protein of smooth muscle and nonmuscle cells.
    Hybridoma. 1988 Jun;7(3):273-88 PMID: 3294163
  53. A miniature flow cell designed for rapid exchange of media under high-power microscope objectives.
    J Gen Microbiol. 1984 Nov;130(11):2915-20 PMID: 6396378
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.109.3.1141
Published
1989-09-00
Pages
1141-52
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2115770
Subset
IM
Grants
NIGMS NIH HHS · GM40580 · United States
NICHD NIH HHS · HD18577 · United States
External Links
PubMed source DOI: 10.1083/jcb.109.3.1141 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2