IngentaConnect EB1 immunofluorescence reveals an increase in growing astral micr...

Authors: Morrison E.E.¹; Askham J.M.¹

Source: European Journal of Cell Biology, Volume 80, Number 12, December 2001 , pp. 749-753(5)

Abstract:

Spindle positioning in animal cells is thought to rely upon the interaction of astral microtubules with the cell cortex. Information on the dynamics of astral microtubules during this process is scarce, in part because of the difficulty in visualising these microtubules by light microscopy. EB1 is a protein which specifically localises to growing microtubule distal tips. Immunostaining for EB1 therefore represents a powerful method for visualising the distribution of growing microtubule tips within cells. In this study we used EB1 immunostaining in mitotic NRK-52E cells to quantitatively analyse the length and number of growing astral microtubules during metaphase and anaphase. We observed a dramatic increase in growing astral microtubule length and number during anaphase. Furthermore, drug treatments which specifically destroyed astral microtubules resulted in an increase in misaligned anaphase but not metaphase spindles. We suggest that an anaphase-specific increase in growing astral microtubule length and number facilitates the maintenance of a correctly aligned spindle in mitotic NRK-52E cells.

Keywords: EB1; microtubules; mitosis; spindle

Language: English

Document Type: Original article

DOI: 10.1078/0171-9335-00221

Affiliations: 1: Molecular Medicine Unit, University of Leeds, St. James's University Hospital, Leeds/UK

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EB1 immunofluorescence reveals an increase in growing astral microtubule length and number during anaphase in NRK-52E cells