| Spindle birefringence of isolated mitotic apparatus: further evidence for two birefringent spindle components. | |
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MedLine Citation:
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PMID: 789386 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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We studied sea-urchin zygote mitotic apparatus (MA) isolated in hexylene glycol, transferred immediately to a glycerol-dimethylsulphoxide medium, and stored for 2 weeks at room temperature. Treatment with 0-5 M KC1 caused loss of 45% of the birefringence, but microtubules remained intact (as seen electron microscopically in glutaraldehyde-fixed MA), and tubulin was not extracted (as determined by polyacrylamide gel electrophoresis). These results suggest that a non-tubulin component which is extracted by the KC1 contributes 45% of the MA birefringence. Further evidence for this conclusion came from indirect immunofluorescence experiments. Non-extracted (control) MA were fixed with formaldehyde and reacted with antibody against tubulin; there was intense staining of the spindle fibres and astral rays. Electron microscopically, however, microtubules were not present in formaldehyde-fixed MA. Since formaldehyde fixation caused breakdown of microtubules but the tubulin remained in the MA (as judged by reaction with antibodies) we suggest that after microtubule breakdown the tubulin remains in the MA because it is bound to a peri-microtubule spindle component (which we call 'substance gamma'). When KCl-extracted MA were fixed with formaldehyde and reacted with antibody against tubulin there was very little staining of spindle fibres and astral rays. Electron microscopically, formaldehyde caused microtubule breakdown, and since the tubulin is lost from formaldehydefixed, KC1-extracted MA (as judged by reaction with antibodies), we suggest that the tubulin-binding component, substance gamma, is extracted by the 0-5 M KC1. Pressure treatment caused the asters not to stain with antibody against tubulin, suggesting that the stability of substance gamma is different in different regions of the mitotic apparatus. |
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Authors:
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A Forer; V I Kalnins; A M Zimmerman |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Journal of cell science Volume: 22 ISSN: 0021-9533 ISO Abbreviation: J. Cell. Sci. Publication Date: 1976 Oct |
Date Detail:
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Created Date: 1976-12-30 Completed Date: 1976-12-30 Revised Date: 2003-11-14 |
Medline Journal Info:
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Nlm Unique ID: 0052457 Medline TA: J Cell Sci Country: ENGLAND |
Other Details:
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Languages: eng Pagination: 115-31 Citation Subset: IM |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Birefringence Cell Fractionation Electrophoresis, Polyacrylamide Gel Female Fluorescent Antibody Technique Microtubules / ultrastructure Mitosis* Organoids / analysis, ultrastructure Potassium Chloride / pharmacology Pressure Sea Urchins Tubulin / analysis Zygote / ultrastructure* |
| Chemical | |
Reg. No./Substance:
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0/Tubulin; 7447-40-7/Potassium Chloride |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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