Microtubules and Ca2+-sequestering membranes in the mitotic apparatus, isolated by a new method

C Petzelt, M Hafner, D Mazia, K W Sawin

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

The mitotic apparatus of sea urchin embryos was isolated using a polyethylene glycol (PEG)/EGTA-medium. Such a procedure preserves the birefringence and the Ca2+ lability of the isolated mitotic apparatus. The method of isolation gives good preservation of the microtubules and of the intracellular Ca2+-transport system as visualized by a monoclonal antibody to a 46-kDa protein. Triple fluorescence studies allow a comparison of the relative locations of microtubules, Ca2+-sequestering membranes and chromatin (by Hoechst 33342) in the mitotic apparatus. We find that the Ca2+-sequestering membranes are concentrated mainly in the centers of the asters and do not follow the distribution of microtubules in the mitotic apparatus. Regulation of microtubules by Ca2+ may not depend on immediate contiguity of microtubules and the Ca2+-regulating sites.
Original languageEnglish
Pages (from-to)268-73
Number of pages6
JournalEuropean journal of cell biology
Volume45
Issue number2
Publication statusPublished - 1988

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