| Teratogen-induced activation of caspase-6 and caspase-7 in early postimplantation mouse embryos. | |
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MedLine Citation:
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PMID: 14686614 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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BACKGROUND: Previous work has shown that teratogens such as hyperthermia (HS), 4-hydroperoxycyclophosphamide (4CP), and staurosporine (ST) induce cell death in day 9 mouse embryos by activating the mitochondrial apoptotic pathway. Key to the activation of this pathway is the activation of a caspase cascade involving the cleavage-induced activation of an initiator procaspase, caspase-9, and the downstream effector procaspase, caspase-3. For example, procaspase-3, an inactive proenzyme of 32 kDa is cleaved by activated caspase-9 to generate a large subunit of approximately 17 kDa and a small subunit of approximately 10 kDa. In turn, caspase-3 is known to target a variety of cellular proteins for proteolytic cleavage as part of the process by which dying cells are eliminated. Previous work has also shown that neuroepithelial cells are sensitive to teratogen-induced activation of this pathway and subsequent cell death whereas cells of the heart are resistant. Although caspase-3 is a key effector caspase activated by teratogens, two other effector caspases, caspase-6 and caspase-7, are known; however, their role in teratogen-induced cell death is unknown. METHODS: Because cleavage-induced generation of specific subunits is the most specific assay for activation of caspases, we have used antibodies that recognize the procaspase and one of its active subunits and a Western blot approach to assess the activation of caspase-6 and caspase-7 in day 9 mouse embryos (or heads, hearts and trunks isolated from whole embryos) exposed to HS, 4CP, and ST. To probe the relationship between teratogen-induced activation of caspase-9/caspase-3 and the activation of caspase-6/caspase-7, we used a mitochondrial-free embryo lysate with or without the addition of cytochrome c, recombinant active caspase-3, or recombinant active caspase-9. RESULTS: Western blot analyses show that these three teratogens, HS, 4CP, and ST, induce the activation of procaspase-6 (appearance of the 13 kDa subunit, p13) and caspase-7 (appearance of the 19 kDa subunit, p19) in day 9 mouse embryos. In vitro studies showed that both caspase-6 and caspase-7 could be activated by the addition of cytochrome c to a lysate prepared from untreated embryos. In addition, caspase-6 could be activated by the addition of either recombinant caspase-3 or caspase-9 to a lysate prepared from untreated embryos. In contrast, caspase-7 could be activated by addition of recombinant caspase-3 but only minimally by recombinant caspase-9. Like caspase-9/caspase-3, caspase-6 and caspase-7 were not activated in hearts isolated from embryos exposed to these three teratogens. CONCLUSIONS: HS, 4CP and ST induce the cleavage-dependent activation of caspase-6 and caspase-7 in day 9 mouse embryos. Results using DEVD-CHO, a caspase-3 inhibitor, suggest that teratogen-induced activation of caspase-6 is mediated by caspase-3. In addition, our data suggest that caspase-7 is activated primarily by caspase-3; however, we cannot rule out the possibility that this caspase is also activated by caspase-9. Finally, we also show that teratogen-induced activation of caspase-6 and caspase-7 are blocked in the heart, a tissue resistant to teratogen-induced cell death. |
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Authors:
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S A Little; W K Kim; P E Mirkes |
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Publication Detail:
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Type: Journal Article; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Cell biology and toxicology Volume: 19 ISSN: 0742-2091 ISO Abbreviation: Cell Biol. Toxicol. Publication Date: 2003 Aug |
Date Detail:
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Created Date: 2003-12-22 Completed Date: 2004-07-20 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 8506639 Medline TA: Cell Biol Toxicol Country: Netherlands |
Other Details:
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Languages: eng Pagination: 215-26 Citation Subset: IM |
Affiliation:
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Birth Defects Research Laboratory, Division of Genetics and Development, Department of Pediatrics, University of Washington, Seattle, WA 98195, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Apoptosis Blastocyst / drug effects* Blotting, Western Caspase 6 Caspase 7 Caspases / metabolism* Cyclophosphamide / analogs & derivatives* Cytochromes c / metabolism Enzyme Activation Enzyme Inhibitors / pharmacology Fever Mice Models, Biological Oligopeptides / pharmacology Recombinant Proteins / chemistry Staurosporine / pharmacology Teratogens* |
| Grant Support | |
ID/Acronym/Agency:
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2R01ES07026/ES/NIEHS NIH HHS; 5P30ES0733/ES/NIEHS NIH HHS; 5R01ES08744/ES/NIEHS NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Enzyme Inhibitors; 0/Oligopeptides; 0/Recombinant Proteins; 0/Teratogens; 0/aspartyl-glutamyl-valyl-aspartal; 39800-16-3/perfosfamide; 50-18-0/Cyclophosphamide; 62996-74-1/Staurosporine; 9007-43-6/Cytochromes c; EC 3.4.22.-/Casp6 protein, mouse; EC 3.4.22.-/Casp7 protein, mouse; EC 3.4.22.-/Caspase 6; EC 3.4.22.-/Caspase 7; EC 3.4.22.-/Caspases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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