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Phallacidin stains the kinetochore region in the mitotic spindle of the green algaeOedogonium spp.

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Summary

We found previously that in living cells ofOedogonium cardiacum andO. donnellii, mitosis is blocked by the drug cytochalasin D (CD). We now report on the staining observed in these spindles with fluorescently actin-labeling reagents, particularly Bodipy FL phallacidin. Normal mitotic cells exhibited spots of staining associated with chromosomes; frequently the spots appeared in pairs during prometaphase-metaphase. During later anaphase and telophase, the staining was confined to the region between chromosomes and poles. The texture of the staining appeared to be somewhat dispersed by CD treatment but it was still present, particularly after shorter (<2 h) exposure. Electron microscopy of CD-treated cells revealed numerous spindle microtubules (MTs); many kinetochores had MTs associated with them, often laterally and some even terminating in the kinetochore as normal, but the usual bundle of kinetochore MTs was never present. As treatment with CD became prolonged, the kinetochores became shrunken and sunk into the chromosomes. These results support the possibility that actin is present in the kinetochore ofOedogonium spp. The previous observations on living cells suggest that it is a functional component of the kinetochore-MT complex involved in the correct attachment of chromosomes to the spindle.

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Abbreviations

CD:

cytochalasin D

EM:

electron microscopy

MBS:

m-maleimidobenzoyl N-hydroxysuccinimide ester

MTs:

microtubules

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  1. School of Botany, University of Melbourne, 3052, Parkville, Vic., Australia

    K. Sampson & J. D. Pickett-Heaps

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  1. K. Sampson
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  2. J. D. Pickett-Heaps
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Sampson, K., Pickett-Heaps, J.D. Phallacidin stains the kinetochore region in the mitotic spindle of the green algaeOedogonium spp.. Protoplasma 217, 166–176 (2001). https://doi.org/10.1007/BF01283397

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