| Identification of potassium-dependent and -independent components of the apoptotic machinery in mouse ovarian germ cells and granulosa cells. | |
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MedLine Citation:
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PMID: 11058539 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Recent studies with thymocytes have suggested a critical role for intracellular potassium in the regulation of apoptosis. In this study, we examined the pathways of K(+) regulation during ovarian cell death. In initial studies, fluorographic analysis demonstrated a significant loss of K(+) during apoptosis stimulated by doxorubicin in oocytes and trophic hormone deprivation in granulosa cells. In oocytes, suppression of potassium efflux by potassium-enriched medium prevented condensation, budding, and fragmentation, although it did not block DNA degradation, suggesting the existence of potassium-independent nucleases in oocytes. Culture of granulosa cells in potassium-enriched medium inhibited internucleosomal DNA cleavage, although high-molecular weight DNA cleavage was apparent, suggesting that the nuclease or nucleases responsible for generating 50-kilobase (kb) fragments in these cells is potassium independent. To address this directly, isolated granulosa cell nuclei were stimulated to autodigest their DNA, and internucleosomal, but not large-fragment, cleavage was completely blocked by 150 mM potassium. We next examined whether the proapoptotic caspases are targets for potassium regulation. In cell-free assays, processing of pro-interleukin-1beta and proteolysis of cellular actin by recombinant caspase-1 and caspase-3, respectively, were suppressed by the presence of 150 mM potassium. Other monovalent ions (NaCl, LiCl) exerted a similar effect in these cell-free assays. Thus, in oocytes and granulosa cells, potassium efflux appears to occur early in the cell death program and may regulate a number of apoptotic events including caspase activity and internucleosomal DNA cleavage. However, there also exist novel potassium-independent pathways in both ovarian germ cells and somatic cells that signal certain apoptotic events, such as large-fragment DNA cleavage. |
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Authors:
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G I Perez; D V Maravei; A M Trbovich; J A Cidlowski; J L Tilly; F M Hughes |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Biology of reproduction Volume: 63 ISSN: 0006-3363 ISO Abbreviation: Biol. Reprod. Publication Date: 2000 Nov |
Date Detail:
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Created Date: 2000-12-15 Completed Date: 2000-12-15 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 0207224 Medline TA: Biol Reprod Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 1358-69 Citation Subset: IM |
Affiliation:
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Vincent Center for Reproductive Biology, Department of Obstetrics and Gynecology, Massachusetts General Hospital/Harvard Medical School, Boston, Massachusetts 02114, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Apoptosis / physiology* Caspases / metabolism Cell Nucleus / metabolism Cytoplasm / metabolism DNA / chemistry, genetics Deoxyribonuclease I / metabolism Electrophoresis, Agar Gel Electrophoresis, Gel, Pulsed-Field Female Germ Cells / physiology* Granulosa Cells / physiology* In Situ Nick-End Labeling Mice Oocytes / physiology Ovarian Follicle / cytology Ovary / cytology, physiology* Plasmids / chemistry, genetics Potassium / physiology* Rats Rats, Sprague-Dawley |
| Grant Support | |
ID/Acronym/Agency:
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R01-AG12279/AG/NIA NIH HHS; R01-ES08430/ES/NIEHS NIH HHS; R01-HD34226/HD/NICHD NIH HHS |
| Chemical | |
Reg. No./Substance:
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7440-09-7/Potassium; 9007-49-2/DNA; EC 3.1.21.1/Deoxyribonuclease I; EC 3.4.22.-/Caspases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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