A Functional in Vitro Model for Studies of Intracellular Motility in Digitonin-permeabilized Erythrophores

Overview

Journal J Cell Biol

Specialty Cell Biology

Date 1982 Sep 1

PMID 6215414

Citations 13

Authors

M E Stearns

R L Ochs

Affiliations

Soon will be listed here.

Abstract

Phase contrast cine results demonstrate that erythrophores maintain saltatory particle motion for hours after permeabilization with 0.001% digitonin in a cytoskeletal stabilizing solution at 23 degrees C. High voltage electron microscopy (HVEM) studies reveal that cytoskeletal elements are retained intact, except in immediate subplasmalemmal regions where the plasma membrane is punctured by digitonin. During digitonin treatments, cells are permeable to ions, small molecules, and antibodies. We find that motion is Ca2+ and ATP-sensitive, and optimal in PIPES buffer (pH 7.2 containing 1 mM Mg2+/ATP and EGTA-CA2+ (10(-7) M Ca2+) at 37 degrees C. Experiments testing the inhibitory effects of vanadate (0.4-10 microM), ouabain (100-600 microM), N-ethyl maleimide, and the cytochalasins B and D indicate that a dyneinlike ATPase may provide the motive force for driving saltatory pigment motion in erythropores.

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References

Fiskum G, Craig S, Decker G, Lehninger A . The cytoskeleton of digitonin-treated rat hepatocytes. Proc Natl Acad Sci U S A. 1980; 77(6):3430-4. PMC: 349630. DOI: 10.1073/pnas.77.6.3430. View

Obika M, Menter D, Tchen T, Taylor J . Actin microfilaments in melanophores of Fundulus heteroclitus. Their possible involvement in melanosome migration. Cell Tissue Res. 1978; 193(3):387-97. DOI: 10.1007/BF00225337. View

SCALLEN T, DIETERT S . The quantitative retention of cholesterol in mouse liver prepared for electron microscopy by fixation in a digitonin-containing aldehyde solution. J Cell Biol. 1969; 40(3):802-13. PMC: 2107645. DOI: 10.1083/jcb.40.3.802. View

Gibbons I, Cosson M, Evans J, GIBBONS B, Houck B, Martinson K . Potent inhibition of dynein adenosinetriphosphatase and of the motility of cilia and sperm flagella by vanadate. Proc Natl Acad Sci U S A. 1978; 75(5):2220-4. PMC: 392523. DOI: 10.1073/pnas.75.5.2220. View

Cande W, McDonald K, Meeusen R . A permeabilized cell model for studying cell division: a comparison of anaphase chromosome movement and cleavage furrow constriction in lysed PtK1 cells. J Cell Biol. 1981; 88(3):618-29. PMC: 2112770. DOI: 10.1083/jcb.88.3.618. View

Ebashi S, Endo M, OTSUKI I . Control of muscle contraction. Q Rev Biophys. 1969; 2(4):351-84. DOI: 10.1017/s0033583500001190. View

Stossel T, Hartwig J . Interactions of actin, myosin, and a new actin-binding protein of rabbit pulmonary macrophages. II. Role in cytoplasmic movement and phagocytosis. J Cell Biol. 1976; 68(3):602-19. PMC: 2109643. DOI: 10.1083/jcb.68.3.602. View

Cantley Jr L, Josephson L, Warner R, Yanagisawa M, Lechene C, Guidotti G . Vanadate is a potent (Na,K)-ATPase inhibitor found in ATP derived from muscle. J Biol Chem. 1977; 252(21):7421-3. View

Cande W . A permeabilized cell model for studying cytokinesis using mammalian tissue culture cells. J Cell Biol. 1980; 87(2 Pt 1):326-35. PMC: 2110752. DOI: 10.1083/jcb.87.2.326. View

10.

Luby K, PORTER K . The control of pigment migration in isolated erythrophores of Holocentrus ascensionis (Osbeck). I. Energy requirements. Cell. 1980; 21(1):13-23. DOI: 10.1016/0092-8674(80)90110-5. View

11.

Connolly J, Kalnins V . Tau and HMW microtubule-associated proteins have different microtubule binding sites in vivo. Eur J Cell Biol. 1980; 21(3):296-300. View

12.

Filo R, BOHR D, Ruegg J . GLYCERINATED SKELETAL AND SMOOTH MUSCLE: CALCIUM AND MAGNESIUM DEPENDENCE. Science. 1965; 147(3665):1581-3. DOI: 10.1126/science.147.3665.1581. View

13.

Bessen M, Fay R, Witman G . Calcium control of waveform in isolated flagellar axonemes of Chlamydomonas. J Cell Biol. 1980; 86(2):446-55. PMC: 2111489. DOI: 10.1083/jcb.86.2.446. View

14.

Hitchcock S . Regulation of motility in nonmuscle cells. J Cell Biol. 1977; 74(1):1-15. PMC: 2109870. DOI: 10.1083/jcb.74.1.1. View

15.

Connolly J, Kalnins V, Cleveland D, Kirschner M . Intracellular localization of the high molecular weight microtubule accessory protein by indirect immunofluorescence. J Cell Biol. 1978; 76(3):781-6. PMC: 2110001. DOI: 10.1083/jcb.76.3.781. View

16.

Schliwa M, Euteneuer U, Herzog W, Weber K . Evidence for rapid structural and functional changes of the melanophore microtubule-organizing center upon pigment movements. J Cell Biol. 1979; 83(3):623-32. PMC: 2110514. DOI: 10.1083/jcb.83.3.623. View

17.

Beckerle M, PORTER K . Inhibitors of dynein activity block intracellular transport in erythrophores. Nature. 1982; 295(5851):701-3. DOI: 10.1038/295701a0. View

18.

Wolosewick J, PORTER K . Microtrabecular lattice of the cytoplasmic ground substance. Artifact or reality. J Cell Biol. 1979; 82(1):114-39. PMC: 2110423. DOI: 10.1083/jcb.82.1.114. View

19.

Connolly J, Kalnins V . The distribution of tau and HMW microtubule-associated proteins in different cell types. Exp Cell Res. 1980; 127(2):341-50. DOI: 10.1016/0014-4827(80)90439-5. View

20.

Schliwa M, Bereiter-Hahn J . Pigment movements in fish melanophores: morphological and physiological studies. V. Evidence for a microtubule-independent contractile system. Cell Tissue Res. 1975; 158(1):61-73. DOI: 10.1007/BF00219951. View