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Protoplasma

, Volume 131, Issue 1, pp 47–59 | Cite as

Metabolic inhibitors and mitosis: I. Effects of dinitrophenol/deoxyglucose and nocodazole on the live spindle

  • T. P. Spurck
  • J. D. Pickett-Heaps
  • M. W. Klymkowsky
Article

Summary

Dinitrophenol and deoxyglucose (DNP/DOG) were used to investigate the effects of ATP depletion on mitotic PtK1 cells. Direct determination of cellular ATP levels showed that the drop of ATP induced by DNP/DOG was rapid; recovery to normal ATP levels was equally rapid once DNP/DOG was removed. On addition of DNP/DOG to live cells, cytoplasmic activity ceased; interphase and prophase cells showed little other response to DNP/DOG. During prometaphase, DNP/DOG induced a pronounced movement of oscillating, monopolar chromosomes towards the spindle poles. As chromosomes became bipolarly attached, DNP/DOG caused the spindle poles themselves to move together. By metaphase, DNP/DOG-treatment led to significant shortening of the spindle which remained intact. DNP/DOG rapidly stopped anaphase chromosome movement and cytokinesis.

Nocodazole (NOC) caused the rapid breakdown of the mitotic spindle; prometaphase chromosomes clustered at the poles and in metaphase cells, the poles were drawn towards the chromosomes as the spindle became disorganized. When cells were pretreated with DNP/DOG and then NOC/DNP/DOG, nocodazole did not break down the spindle. When nocodazole was applied first to break down spindle MTs then DNP/DOG was added to the nocodazole, a second contraction was often induced by the DNP/DOG in the absence of spindle microtubules (MTs). Chromosomes expanded appreciably outwards from the poles when the DNP/DOG was removed, even when the cells remained in nocodazole.

Keywords

Mitosis ATP Metabolic inhibitors Spindle Nocodazole 

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Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • T. P. Spurck
    • 1
  • J. D. Pickett-Heaps
    • 1
  • M. W. Klymkowsky
    • 1
  1. 1.Department of Molecular, Cellular and Developmental BiologyUniversity of ColoradoBoulderUSA

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