| Isolation of skeletal muscle nuclei. | |
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MedLine Citation:
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PMID: 4287141 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A method employing aqueous media for isolation of nuclei from rat skeletal muscle is described. The technique involves (a) mincing and then homogenizing in a 0.32 M sucrose-salt solution with a Potter-Elvehjem type homogenizer using a Delrin (an acetal resin) pestle and a carefully controlled, relatively large pestle-to-glass clearance, (b) filtering through fiberglass and stainless steel screens of predetermined mesh size to remove myofibrils and connective tissue, and (c) centrifuging in a 2.15 M sucrose-salt solution containing 0.7 mM ATP. Electron and phase-contrast microscopic observations show that the nuclei are intact, unencumbered by cytoplasmic tags, and possess well preserved distinct nucleoli, nucleoplasm, and nuclear membranes. Cytoplasmic contamination is minimal and mainly mitochondrial. Chemical assays of the nuclear fraction show that the DNA/protein and RNA/DNA ratios are comparable to those obtained in other tissues. These ratios, as well as the low specific activity obtained for cytochrome c oxidase and the virtual absence of myofibrillar ATPase, indicate a high degree of purity with minimal mitochondrial and myofibrillar contamination. The steps comprising the technique and the reasons for their selection are discussed. |
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Authors:
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J C Edelman; P M Edelman; K M Kniggee; I L Schwartz |
Publication Detail:
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Type: In Vitro; Journal Article |
Journal Detail:
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Title: The Journal of cell biology Volume: 27 ISSN: 0021-9525 ISO Abbreviation: J. Cell Biol. Publication Date: 1965 Nov |
Date Detail:
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Created Date: 1966-09-02 Completed Date: 1966-09-02 Revised Date: 2010-06-22 |
Medline Journal Info:
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Nlm Unique ID: 0375356 Medline TA: J Cell Biol Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 365-77 Citation Subset: IM |
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| MeSH Terms | |
Descriptor/Qualifier:
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Adenosine Triphosphatases Adenosine Triphosphate Animals Cell Nucleus* Centrifugation DNA Electron Transport Complex IV Filtration Histological Techniques* Microscopy, Electron Microscopy, Phase-Contrast Mitochondria Muscles / cytology* Myofibrils Nucleoproteins RNA Rats Sodium Chloride Subcellular Fractions Sucrose |
| Chemical | |
Reg. No./Substance:
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0/Nucleoproteins; 56-65-5/Adenosine Triphosphate; 57-50-1/Sucrose; 63231-63-0/RNA; 7647-14-5/Sodium Chloride; 9007-49-2/DNA; EC 1.9.3.1/Electron Transport Complex IV; EC 3.6.1.-/Adenosine Triphosphatases |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
| Full Text | |
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Journal Information Journal ID (nlm-ta): J Cell Biol ISSN: 0021-9525 ISSN: 1540-8140 Publisher: The Rockefeller University Press |
Article Information Download PDF  Received Day: 14 Month: 6 Year: 1965 Print publication date: Day: 1 Month: 11 Year: 1965 Volume: 27 Issue: 2 First Page: 365 Last Page: 378 ID: 2106724 PubMed Id: 4287141 |
| ISOLATION OF SKELETAL MUSCLE NUCLEI | |
| Jean C. Edelman | |
| P. Michael Edelman | |
| Karl M. Knigge | |
| Irving L. Schwartz | |
| From the Departments of Physiology and Anatomy, University of Cincinnati College of Medicine, Cincinnati, Ohio, and The Medical Research Center, Brookhaven National Laboratory, Upton, New York. |
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| Dr. Knigge's present address is the Department of Anatomy, University of Rochester School of Medicine and Dentistry, Rochester, New York |
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