Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
QUANTITATIVE-DETERMINATION OF THE PROPORTION OF MICROTUBULE POLYMER PRESENT DURING THE MITOSIS-INTERPHASE TRANSITION
Autore:
ZHAI Y; BORISY GG;
Indirizzi:
UNIV WISCONSIN,MOLEC BIOL LAB MADISON WI 53706
Titolo Testata:
Journal of Cell Science
, volume: 107, anno: 1994,
parte:, 4
pagine: 881 - 890
SICI:
0021-9533(1994)107:<881:QOTPOM>2.0.ZU;2-N
Fonte:
ISI
Lingua:
ENG
Soggetto:
KINETOCHORE MICROTUBULES; CYTOPLASMIC DYNEIN; ASSEMBLY INVITRO; MITOTIC SPINDLES; CELLS; TUBULIN; DYNAMICS; ATTACHMENT; NOCODAZOLE; TAXOL;
Keywords:
TUBULIN; MICROTUBULE; INTERPHASE; MITOSIS; FLUORESCENCE MICROSCOPY;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
41
Recensione:
Indirizzi per estratti:
Citazione:
Y. Zhai e G.G. Borisy, "QUANTITATIVE-DETERMINATION OF THE PROPORTION OF MICROTUBULE POLYMER PRESENT DURING THE MITOSIS-INTERPHASE TRANSITION", Journal of Cell Science, 107, 1994, pp. 881-890

Abstract

We have developed a new method for determining levels of tubulin polymer, based on quantitative fluorescence detection of x-rhodamine tubulin microinjected into living cells and we have applied this method to analysis of the mitosis-interphase transition. LLC-PK cells in interphase and mitosis were microinjected, then cooled and rewarmed to drive tubulin incorporation. Total tubulin fluorescence in individual, living cells was quantified using a cooled, scientific grade CCD image sensor. Cells were then washed and lysed into a microtubule-stabilizing buffer to extract the soluble pool. Total tubulin polymer fluorescence was determined for the extracted cells in the same way as for living cells. Fluorescence images were corrected by flat-fielding and background subtraction. The ratio of extracted cell fluorescence/living cell fluorescence for individual cells, was taken as the proportion of tubulin as polymer. Cells in M-phase, G(1) and random interphase were analyzed. G(1) cells had almost the same proportion as random interphase cells. Mitotic cells gave a value of 90+/-5% of G(1) cells at 37 degrees C. Within M-phase, levels of tubulin as polymer in metaphase and earlyanaphase were not significantly different. In contrast to the generalexpectation of microtubule depolymerization at anaphase onset, these results indicate that as cells exit mitosis, the overall proportion oftubulin as polymer does not change dramatically even though the mitotic spindle disassembles. We conclude that the mitosis-interphase transition is accompanied by a redistribution of tubulin at an essentially constant polymer level. Therefore, a global shift to depolymerization conditions is not the driving force for anaphase chromosome movement.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 27/09/20 alle ore 21:43:24