Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1974 Jul 1;62(1):164–174. doi: 10.1083/jcb.62.1.164

MICROTUBULE PROTEIN DURING CILIOGENESIS IN THE MOUSE OVIDUCT

Ilona Staprans 1, Ellen Roter Dirksen 1
PMCID: PMC2109194  PMID: 4407047

Abstract

A colchicine-binding assay and quantitative sodium dodecyl sulfate gel electrophoresis have been used to determine the changes which occur in microtubule protein (tubulin) concentrations in the particulate and soluble fractions of mouse oviduct homogenates during that period of development when centriole formation and cilium formation are at a maximum. When mouse oviducts, at various ages after birth, are homogenized in Tris-sucrose buffer, tubulin concentration is partitioned between the soluble (70%) and particulate (30%) fractions. During the period of most active organelle formation (3–12 days), there is a marked increase in colchicine-binding specific activity, in both the soluble and particulate fractions. Microtubule protein concentration increases from 16 to 24% in the soluble fraction, declining to 14% in the adult. In the particulate fractions, microtubule protein concentration increases from 16 to 27%, leveling off at 16% in the adult. We have concluded from these observations and from electron microscopy that colchicine-binding activity in the particulate fractions is related to the presence of centriole precursors in the pellets of homogenized oviducts from newborn mice. These data further suggest that centriole precursor structures are conveniently packaged aggregates of microtubule protein actively synthesized between 3 and 5 days, and maintained at a maximum during the most active period of organelle assembly.

Full Text

The Full Text of this article is available as a PDF (1,012.6 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson R. G., Brenner R. M. The formation of basal bodies (centrioles) in the Rhesus monkey oviduct. J Cell Biol. 1971 Jul;50(1):10–34. doi: 10.1083/jcb.50.1.10. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Auclair W., Siegel B. W. Cilia regeneration in the sea urchin embryo: evidence for a pool of ciliary proteins. Science. 1966 Nov 18;154(3751):913–915. doi: 10.1126/science.154.3751.913. [DOI] [PubMed] [Google Scholar]
  3. Bamburg J. R., Shooter E. M., Wilson L. Assay of microtuble protein in embryonic chick dorsal root ganglia. Neurobiology. 1973;3(3):162–173. [PubMed] [Google Scholar]
  4. Bamburg J. R., Shooter E. M., Wilson L. Developmental changes in microtubule protein of chick brain. Biochemistry. 1973 Apr 10;12(8):1476–1482. doi: 10.1021/bi00732a002. [DOI] [PubMed] [Google Scholar]
  5. Borisy G. G. A rapid method for quantitative determination of microtubule protein using DEAE-cellulose filters. Anal Biochem. 1972 Dec;50(2):373–385. doi: 10.1016/0003-2697(72)90046-2. [DOI] [PubMed] [Google Scholar]
  6. Borisy G. G., Olmsted J. B., Klugman R. A. In vitro aggregation of cytoplasmic microtubule subunits. Proc Natl Acad Sci U S A. 1972 Oct;69(10):2890–2894. doi: 10.1073/pnas.69.10.2890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Borisy G. G., Olmsted J. B. Nucleated assembly of microtubules in porcine brain extracts. Science. 1972 Sep 29;177(4055):1196–1197. doi: 10.1126/science.177.4055.1196. [DOI] [PubMed] [Google Scholar]
  8. Borisy G. G., Taylor E. W. The mechanism of action of colchicine. Binding of colchincine-3H to cellular protein. J Cell Biol. 1967 Aug;34(2):525–533. doi: 10.1083/jcb.34.2.525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. DIRKSEN E. R. The presence of centrioles in artificially activated sea urchin eggs. J Biophys Biochem Cytol. 1961 Oct;11:244–247. doi: 10.1083/jcb.11.1.244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dippell R. V. The development of basal bodies in paramecium. Proc Natl Acad Sci U S A. 1968 Oct;61(2):461–468. doi: 10.1073/pnas.61.2.461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dirksen E. R. Centriole morphogenesis in developing ciliated epithelium of the mouse oviduct. J Cell Biol. 1971 Oct;51(1):286–302. doi: 10.1083/jcb.51.1.286. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dirksen E. R., Satir P. Ciliary activity in the mouse oviduct as studied by transmission and scanning electron microscopy. Tissue Cell. 1972;4(3):389–403. doi: 10.1016/s0040-8166(72)80017-x. [DOI] [PubMed] [Google Scholar]
  13. Feit H., Barondes S. H. Colchicine-binding activity in particulate fractions of mouse brain. J Neurochem. 1970 Sep;17(9):1355–1364. doi: 10.1111/j.1471-4159.1970.tb06870.x. [DOI] [PubMed] [Google Scholar]
  14. Frigon R. P., Lee J. C. The stabilization of calf-brain microtubule protein by sucrose. Arch Biochem Biophys. 1972 Dec;153(2):587–589. doi: 10.1016/0003-9861(72)90376-1. [DOI] [PubMed] [Google Scholar]
  15. Frisch D., Farbman A. I. Development of order during ciliogenesis. Anat Rec. 1968 Oct;162(2):221–232. doi: 10.1002/ar.1091620209. [DOI] [PubMed] [Google Scholar]
  16. Inoué S., Sato H. Cell motility by labile association of molecules. The nature of mitotic spindle fibers and their role in chromosome movement. J Gen Physiol. 1967 Jul;50(6 Suppl):259–292. [PMC free article] [PubMed] [Google Scholar]
  17. Kalnins V. I., Chung C. K., Turnbull C. Procentrioles in ciliating and ciliated cells of chick trachea. Z Zellforsch Mikrosk Anat. 1972;135(4):461–471. doi: 10.1007/BF00583430. [DOI] [PubMed] [Google Scholar]
  18. Kalnins V. I., Porter K. R. Centriole replication during ciliogenesis in the chick tracheal epithelium. Z Zellforsch Mikrosk Anat. 1969;100(1):1–30. doi: 10.1007/BF00343818. [DOI] [PubMed] [Google Scholar]
  19. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  20. Outka D. E., Kluss B. C. The ameba-to-flagellate transformation in Tetramitus rostratus. II. Microtubular morphogenesis. J Cell Biol. 1967 Nov;35(2):323–346. doi: 10.1083/jcb.35.2.323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Raff R. A., Greenhouse G., Gross K. W., Gross P. R. Synthesis and storage of microtubule proteins by sea urchin embryos. J Cell Biol. 1971 Aug;50(2):516–527. doi: 10.1083/jcb.50.2.516. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Raff R. A., Kaumeyer J. F. Soluble microtubule proteins of the sea urchin embryo: partial characterization of the proteins and behavior of the pool in early development. Dev Biol. 1973 Jun;32(2):309–320. doi: 10.1016/0012-1606(73)90243-1. [DOI] [PubMed] [Google Scholar]
  23. Rosenbaum J. L., Child F. M. Flagellar regeneration in protozoan flagellates. J Cell Biol. 1967 Jul;34(1):345–364. doi: 10.1083/jcb.34.1.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rosenbaum J. L., Moulder J. E., Ringo D. L. Flagellar elongation and shortening in Chlamydomonas. The use of cycloheximide and colchicine to study the synthesis and assembly of flagellar proteins. J Cell Biol. 1969 May;41(2):600–619. doi: 10.1083/jcb.41.2.600. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Shelanski M. L., Gaskin F., Cantor C. R. Microtubule assembly in the absence of added nucleotides. Proc Natl Acad Sci U S A. 1973 Mar;70(3):765–768. doi: 10.1073/pnas.70.3.765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Spurr A. R. A low-viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res. 1969 Jan;26(1):31–43. doi: 10.1016/s0022-5320(69)90033-1. [DOI] [PubMed] [Google Scholar]
  27. Steinman R. M. Inhibitory effects of colchicine on ciliogenesis in ectoderm of Xenopus laevis. J Ultrastruct Res. 1970 Feb;30(3):423–440. doi: 10.1016/s0022-5320(70)80073-9. [DOI] [PubMed] [Google Scholar]
  28. Tamura S. Synthesis and assembly of microtubule proteins in Tetrahymena pyriformis. Exp Cell Res. 1971 Sep;68(1):180–185. doi: 10.1016/0014-4827(71)90601-x. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Weisenberg R. C., Borisy G. G., Taylor E. W. The colchicine-binding protein of mammalian brain and its relation to microtubules. Biochemistry. 1968 Dec;7(12):4466–4479. doi: 10.1021/bi00852a043. [DOI] [PubMed] [Google Scholar]
  31. Weisenberg R. C. Microtubule formation in vitro in solutions containing low calcium concentrations. Science. 1972 Sep 22;177(4054):1104–1105. doi: 10.1126/science.177.4054.1104. [DOI] [PubMed] [Google Scholar]
  32. Williams N. E., Nelsen E. M. Regulation of microtubules in Tetrahymena. II. Relation between turnover of microtubule proteins and microtubule dissociation and assembly during oral replacement. J Cell Biol. 1973 Feb;56(2):458–465. doi: 10.1083/jcb.56.2.458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wilson L., Friedkin M. The biochemical events of mitosis. II. The in vivo and in vitro binding of colchicine in grasshopper embryos and its possible relation to inhibition of mitosis. Biochemistry. 1967 Oct;6(10):3126–3135. doi: 10.1021/bi00862a021. [DOI] [PubMed] [Google Scholar]
  34. Wilson L., Meza I. The mechanism of action of colchicine. Colchicine binding properties of sea urchin sperm tail outer doublet tubulin. J Cell Biol. 1973 Sep;58(3):709–719. doi: 10.1083/jcb.58.3.709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wilson L. Properties of colchicine binding protein from chick embryo brain. Interactions with vinca alkaloids and podophyllotoxin. Biochemistry. 1970 Dec 8;9(25):4999–5007. doi: 10.1021/bi00827a026. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES