Different reactivity with monoclonal anti-tubulin antibodies between native and fixed mitotic microtubules in sea urchin eggs

M. T. Oka, T. Arai, Y. Hamaguchi

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The effect on fixation on the reactivities of mitotic microtubules with monoclonal anti-tubulin antibodies was investigated by the indirect immunofluorescence procedure. All of the seven antibodies used intensely stained mitotic microtubules in sea urchin eggs lysed and fixed with methanol at -20°C, whereas only two of them stained the stabilized microtubules in the lysed eggs before the fixation. The other five did not stain the mitotic microtubules even after microtubule components other than tubulin were removed by treating the lysed eggs with 0.4 M KCl solution containing taxol. These results exclude the possibility that the fixation affects proteins, which interact with microtubules including microtubule-associated proteins (MAPs) and interfere with the binding of monoclonal antibodies with tubulin, and strongly suggest that the fixation directly affects the three-dimensional conformation of tubulin. Furthermore, microinjection of these antibodies indicated the results as follows [combining the results reported previously; Oka et al., 1990: Cell Struct. Funct. 15: 373-378]: The antibodies which stained mitotic microtubules stabilized in the lysed eggs induced disassembly of native mitotic microtubules in the living eggs, but those which did not stain the stabilized microtubules did not disassemble the native microtubules. From these results, it is suggested that the monoclonal antibodies which stain microtubules in the eggs lysed but not fixed are useful for microinjection experiments.

Original languageEnglish
Pages (from-to)241-249
Number of pages9
JournalCell Motility and the Cytoskeleton
Volume29
Issue number3
DOIs
Publication statusPublished - 1994

Fingerprint

Sea Urchins
Tubulin
Microtubules
Eggs
Anti-Idiotypic Antibodies
Monoclonal Antibodies
Coloring Agents
Microinjections
Antibodies
Microtubule-Associated Proteins
Indirect Fluorescent Antibody Technique
Paclitaxel
Methanol

Keywords

  • immunofluorescence
  • microinjection
  • mitotic apparatus
  • monoclonal antibodies
  • sand dollar egg
  • tubulin isotypes

ASJC Scopus subject areas

  • Cell Biology

Cite this

Different reactivity with monoclonal anti-tubulin antibodies between native and fixed mitotic microtubules in sea urchin eggs. / Oka, M. T.; Arai, T.; Hamaguchi, Y.

In: Cell Motility and the Cytoskeleton, Vol. 29, No. 3, 1994, p. 241-249.

Research output: Contribution to journalArticle

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