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. 1980 Aug;66(2):284–291. doi: 10.1172/JCI109855

Platelets and Microtubules

EFFECT OF COLCHICINE AND D2O ON PLATELET AGGREGATION AND RELEASE INDUCED BY CALCIUM IONOPHORE A23187

D Menche 1, A Israel 1, S Karpatkin 1
PMCID: PMC371709  PMID: 6772671

Abstract

We examined the role of microtubules in platelet aggregation and secretion (release reaction) induced by the calcium ionophore A23187 (0.8-5 μM). At these concentrations, platelet aggregation was preceded by a lag period of ∼1 min. Colchicine (an agent that disrupts microtubule assembly-disassembly) was shown to bind to platelet microtubules by employing [3H]colchicine at a concentration that is specific for microtubules in other tissues (10 nM). Colchicine prolonged the lag period, inhibited the secondary wave of platelet aggregation, and inhibited the release reaction (release of [14C]serotonin). Platelets were next incubated with 20-60% D2O, an agent that stabilizes microtubules. D2O overcame colchicine-induced inhibition of the lag period, aggregation, and release. D2O alone enhanced platelet aggregation by 59±14% (SEM) and shortened the lag period by 43±10%. We conclude that functioning microtubules are required for platelet aggregation and release, and that microtubules of platelet preparations are functioning submaximally.

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Selected References

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  1. Andrews P. Estimation of the molecular weights of proteins by Sephadex gel-filtration. Biochem J. 1964 May;91(2):222–233. doi: 10.1042/bj0910222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Behnke O. Further studies on microtubules. A marginal bundle in human and rat thrombocytes. J Ultrastruct Res. 1965 Dec;13(5):469–477. doi: 10.1016/s0022-5320(65)90009-2. [DOI] [PubMed] [Google Scholar]
  3. Boyle Kay M. M., Fudenberg H. H. Inhibition and reversal of platelet activation by cytochalasin B or colcemid. Nature. 1973 Aug 3;244(5414):288–289. doi: 10.1038/244288a0. [DOI] [PubMed] [Google Scholar]
  4. Charo I. F., Feinman R. D., Detwiler T. C. Interrelations of platelet aggregation and secretion. J Clin Invest. 1977 Oct;60(4):866–873. doi: 10.1172/JCI108841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dechatelet L. R., Cooper M. R., McCall C. E. Dissociation by colchicine of the hexose monophosphate shunt activation from the bactericidal activity of the leukocyte. Infect Immun. 1971 Jan;3(1):66–72. doi: 10.1128/iai.3.1.66-72.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ehrlich H. P., Ross R., Bornstein P. Effects of antimicrotubular agents on the secretion of collagen. A biochemical and morphological study. J Cell Biol. 1974 Aug;62(2):390–405. doi: 10.1083/jcb.62.2.390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Feinstein M. B., Fraser C. Human platelet secretion and aggregation induced by calcium ionophores. Inhibition by PGE1 and dibutyryl cyclic AMP. J Gen Physiol. 1975 Nov;66(5):561–581. doi: 10.1085/jgp.66.5.561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Goldfinger S. E., Howell R. R., Seegmiller J. E. Suppression of metabolic accompaniments of phagocytosis by colchicine. Arthritis Rheum. 1965 Dec;8(6):1112–1122. doi: 10.1002/art.1780080610. [DOI] [PubMed] [Google Scholar]
  9. Harris E. D., Jr, Krane S. M. Effects of colchicine on collagenase in cultures of rheumatoid synovium. Arthritis Rheum. 1971 Nov-Dec;14(6):669–684. doi: 10.1002/art.1780140602. [DOI] [PubMed] [Google Scholar]
  10. Krishan A., Hsu D. Vinblastine-induced ribosomal complexes. Effect of some metabolic inhibitors on their formation and structure. J Cell Biol. 1971 Jun;49(3):927–932. doi: 10.1083/jcb.49.3.927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lyons R. M., Stanford N., Majerus P. W. Thrombin-induced protein phosphorylation in human platelets. J Clin Invest. 1975 Oct;56(4):924–936. doi: 10.1172/JCI108172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Margolis R. L., Wilson L. Addition of colchicine--tubulin complex to microtubule ends: the mechanism of substoichiometric colchicine poisoning. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3466–3470. doi: 10.1073/pnas.74.8.3466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Massini P., Lüscher E. F. Some effects of ionophores for divalent cations on blood platelets. Comparison with the effects of thrombin. Biochim Biophys Acta. 1974 Nov 4;372(1):109–121. doi: 10.1016/0304-4165(74)90077-4. [DOI] [PubMed] [Google Scholar]
  14. Mizel S. B., Wilson L. Nucleoside transport in mammalian cells. Inhibition by colchicine. Biochemistry. 1972 Jul 4;11(14):2573–2578. doi: 10.1021/bi00764a003. [DOI] [PubMed] [Google Scholar]
  15. Olmsted J. B., Borisy G. G. Characterization of microtubule assembly in porcine brain extracts by viscometry. Biochemistry. 1973 Oct 9;12(21):4282–4289. doi: 10.1021/bi00745a037. [DOI] [PubMed] [Google Scholar]
  16. Olmsted J. B., Borisy G. G. Microtubules. Annu Rev Biochem. 1973;42:507–540. doi: 10.1146/annurev.bi.42.070173.002451. [DOI] [PubMed] [Google Scholar]
  17. Patzelt C., Brown D., Jeanrenaud B. Inhibitory effect of colchicine on amylase secretion by rat parotid glands. Possible localization in the Golgi area. J Cell Biol. 1977 Jun;73(3):578–593. doi: 10.1083/jcb.73.3.578. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sneddon J. M. Effect of mitosis inhibitors on blood platelet microtubules and aggregation. J Physiol. 1971 Apr;214(1):145–158. doi: 10.1113/jphysiol.1971.sp009424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Soppitt G. D., Mitchell J. R. The effect of colchicine on human platelet behaviour. J Atheroscler Res. 1969 Sep-Oct;10(2):247–252. doi: 10.1016/s0368-1319(69)80012-8. [DOI] [PubMed] [Google Scholar]
  20. Stephens R. E., Edds K. T. Microtubules: structure, chemistry, and function. Physiol Rev. 1976 Oct;56(4):709–777. doi: 10.1152/physrev.1976.56.4.709. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  23. White J. G. Effects of colchicine and Vinca alkaloids on human platelets. I. Influence on platelet microtubules and contractile function. Am J Pathol. 1968 Aug;53(2):281–291. [PMC free article] [PubMed] [Google Scholar]
  24. White J. G. Effects of colchicine and vinca alkaloids on human platelets. 3. Influence on primary internal contraction and secondary aggregation. Am J Pathol. 1969 Mar;54(3):467–478. [PMC free article] [PubMed] [Google Scholar]
  25. White J. G. Fine structural alterations induced in platelets by adenosine diphosphate. Blood. 1968 May;31(5):604–622. [PubMed] [Google Scholar]

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