Protein databases for compacted eight-cell and blastocyst-stage mouse embryos
High‐resolution two‐dimensional sodium dodecyl sulfate‐polyacrylamide (2D‐SDS) gel electrophoresis combined with computerized analysis of gel images was used to construct and analyze protein databases for two stages of preimplantation mouse embryogenesis, the compacted eight‐cell stage and the fully...
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| Veröffentlicht in: | Molecular reproduction and development 1994-01, Vol.37 (1), p.34-47 |
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| creator | Shi, C. Z. Collins, H. W. Garside, W. T. Buettger, C. W. Matschinsky, F. M. Heyner, S. |
| description | High‐resolution two‐dimensional sodium dodecyl sulfate‐polyacrylamide (2D‐SDS) gel electrophoresis combined with computerized analysis of gel images was used to construct and analyze protein databases for two stages of preimplantation mouse embryogenesis, the compacted eight‐cell stage and the fully expanded blastocyst stage. These stages were chosen for their ease in identification of multiple synchronous embryos. Synchronous cohorts of 30–50 embryos were labelled with L‐[35S]methionine for 2 hr. The embryos were then lysed in 30 μl hot SDS sample buffer, and the lysates were stored at −80°C until the gels were run. Five replicates were run for eight‐cell embryos, and four for blastocyst‐stage embryos. The samples were processed for 2D gel electrophoresis and fluorography; multiple exposures were made. Gel images were analyzed using the PDQUEST system, and databases were constructed. Analysis of the databases for both developmental stages showed high reproducibility of protein spots in multiple gel images. Of 1,674 total spots in eight‐cell embryo standards, >79% of spots had a percentage error (S.E.M./average) 45% had a percentage error |
| doi_str_mv | 10.1002/mrd.1080370106 |
| format | Article |
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Z. ; Collins, H. W. ; Garside, W. T. ; Buettger, C. W. ; Matschinsky, F. M. ; Heyner, S.</creator><creatorcontrib>Shi, C. Z. ; Collins, H. W. ; Garside, W. T. ; Buettger, C. W. ; Matschinsky, F. M. ; Heyner, S.</creatorcontrib><description>High‐resolution two‐dimensional sodium dodecyl sulfate‐polyacrylamide (2D‐SDS) gel electrophoresis combined with computerized analysis of gel images was used to construct and analyze protein databases for two stages of preimplantation mouse embryogenesis, the compacted eight‐cell stage and the fully expanded blastocyst stage. These stages were chosen for their ease in identification of multiple synchronous embryos. Synchronous cohorts of 30–50 embryos were labelled with L‐[35S]methionine for 2 hr. The embryos were then lysed in 30 μl hot SDS sample buffer, and the lysates were stored at −80°C until the gels were run. Five replicates were run for eight‐cell embryos, and four for blastocyst‐stage embryos. The samples were processed for 2D gel electrophoresis and fluorography; multiple exposures were made. Gel images were analyzed using the PDQUEST system, and databases were constructed. Analysis of the databases for both developmental stages showed high reproducibility of protein spots in multiple gel images. Of 1,674 total spots in eight‐cell embryo standards, >79% of spots had a percentage error (S.E.M./average) <50%, and >45% had a percentage error <30%. Similarly, of 1,653 total spots in blastocyst‐stage embryo standards, 74% of spots had a percentage error <50%, and approximately 47% of spots had a percentage error <30%. Forty‐three spots (approximately 3% of the total spots) were found to be detected only in the eight‐cell stage, while 75 spots were detected solely in the blastocyst stage. Sixty‐nine proteins showed a greater than threefold increase in isotope incorporation from the eight‐cell to the blastocyst stage, with a percentage error <50% in both the eight‐cell and the blastocyst stages. In contrast, 41 of the proteins showed a decrease during this period. Analysis of the protein databases described in this study has allowed us to document the overall quantitative changes in proteins from the compacted eight‐cell stage to the blastocyst stage of mouse preimplantation development. These databases provide a valuable tool for further detailed quantitative analysis of specific proteins associated with developmental events. In addition they will permit analysis of the effects of environmental factors, such as growth factors, on early embryo development. © 1994 Wiley‐Liss, Inc.</description><identifier>ISSN: 1040-452X</identifier><identifier>EISSN: 1098-2795</identifier><identifier>DOI: 10.1002/mrd.1080370106</identifier><identifier>PMID: 8129929</identifier><identifier>CODEN: MREDEE</identifier><language>eng</language><publisher>Hoboken: Wiley Subscription Services, Inc., A Wiley Company</publisher><subject>Animals ; Biological and medical sciences ; Blastocyst - cytology ; Blastocyst - metabolism ; Blastocyst - physiology ; Cell Division ; Databases, Factual ; Early stages. Segmentation. Gastrulation. Neurulation ; Electrophoresis, Gel, Two-Dimensional ; Electrophoresis, Polyacrylamide Gel ; Embryology: invertebrates and vertebrates. Teratology ; Female ; Fundamental and applied biological sciences. Psychology ; Male ; Methionine - metabolism ; Mice ; Mice, Inbred Strains ; Molecular Weight ; PDQUEST ; Preimplantation development ; Protein Biosynthesis ; Proteins - analysis ; Proteins - isolation & purification ; Two-dimensional gel electrophoresis</subject><ispartof>Molecular reproduction and development, 1994-01, Vol.37 (1), p.34-47</ispartof><rights>Copyright © 1994 Wiley‐Liss, Inc.</rights><rights>1994 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3886-cf4ac91b9f4e4ee0f0fbdf093284f649e5f767da87a2c1834eb97a5a25d423b23</citedby><cites>FETCH-LOGICAL-c3886-cf4ac91b9f4e4ee0f0fbdf093284f649e5f767da87a2c1834eb97a5a25d423b23</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fmrd.1080370106$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fmrd.1080370106$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,4024,27923,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=3925431$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8129929$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Shi, C. Z.</creatorcontrib><creatorcontrib>Collins, H. W.</creatorcontrib><creatorcontrib>Garside, W. T.</creatorcontrib><creatorcontrib>Buettger, C. W.</creatorcontrib><creatorcontrib>Matschinsky, F. M.</creatorcontrib><creatorcontrib>Heyner, S.</creatorcontrib><title>Protein databases for compacted eight-cell and blastocyst-stage mouse embryos</title><title>Molecular reproduction and development</title><addtitle>Mol. Reprod. Dev</addtitle><description>High‐resolution two‐dimensional sodium dodecyl sulfate‐polyacrylamide (2D‐SDS) gel electrophoresis combined with computerized analysis of gel images was used to construct and analyze protein databases for two stages of preimplantation mouse embryogenesis, the compacted eight‐cell stage and the fully expanded blastocyst stage. These stages were chosen for their ease in identification of multiple synchronous embryos. Synchronous cohorts of 30–50 embryos were labelled with L‐[35S]methionine for 2 hr. The embryos were then lysed in 30 μl hot SDS sample buffer, and the lysates were stored at −80°C until the gels were run. Five replicates were run for eight‐cell embryos, and four for blastocyst‐stage embryos. The samples were processed for 2D gel electrophoresis and fluorography; multiple exposures were made. Gel images were analyzed using the PDQUEST system, and databases were constructed. Analysis of the databases for both developmental stages showed high reproducibility of protein spots in multiple gel images. Of 1,674 total spots in eight‐cell embryo standards, >79% of spots had a percentage error (S.E.M./average) <50%, and >45% had a percentage error <30%. Similarly, of 1,653 total spots in blastocyst‐stage embryo standards, 74% of spots had a percentage error <50%, and approximately 47% of spots had a percentage error <30%. Forty‐three spots (approximately 3% of the total spots) were found to be detected only in the eight‐cell stage, while 75 spots were detected solely in the blastocyst stage. Sixty‐nine proteins showed a greater than threefold increase in isotope incorporation from the eight‐cell to the blastocyst stage, with a percentage error <50% in both the eight‐cell and the blastocyst stages. In contrast, 41 of the proteins showed a decrease during this period. Analysis of the protein databases described in this study has allowed us to document the overall quantitative changes in proteins from the compacted eight‐cell stage to the blastocyst stage of mouse preimplantation development. These databases provide a valuable tool for further detailed quantitative analysis of specific proteins associated with developmental events. In addition they will permit analysis of the effects of environmental factors, such as growth factors, on early embryo development. © 1994 Wiley‐Liss, Inc.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Blastocyst - cytology</subject><subject>Blastocyst - metabolism</subject><subject>Blastocyst - physiology</subject><subject>Cell Division</subject><subject>Databases, Factual</subject><subject>Early stages. Segmentation. Gastrulation. Neurulation</subject><subject>Electrophoresis, Gel, Two-Dimensional</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Embryology: invertebrates and vertebrates. Teratology</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Male</subject><subject>Methionine - metabolism</subject><subject>Mice</subject><subject>Mice, Inbred Strains</subject><subject>Molecular Weight</subject><subject>PDQUEST</subject><subject>Preimplantation development</subject><subject>Protein Biosynthesis</subject><subject>Proteins - analysis</subject><subject>Proteins - isolation & purification</subject><subject>Two-dimensional gel electrophoresis</subject><issn>1040-452X</issn><issn>1098-2795</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkM1v00AQxVcVqC2FKzckHxA3l_2yd_eI0jYgWqgQiKqX1Xg927q143RnI8h_j6NEQZw4zZPm92aeHmOvBT8VnMv3Q2onYbkyXPD6gB0L7mwpjauebbTmpa7kzRF7QfTAOXfO8kN2aIV0TrpjdnWdxozdomghQwOEVMQxFWEclhAytgV2d_e5DNj3BSzaoumB8hjWlEvKcIfFMK4ICxyatB7pJXseoSd8tZsn7MfF-ffZx_Ly6_zT7MNlGZS1dRmihuBE46JGjcgjj00buVPS6lhrh1U0tWnBGpBBWKWxcQYqkFWrpWqkOmHvtneXaXxaIWU_dLTJCAuc8nhTK6eFExN4ugVDGokSRr9M3QBp7QX3m_781J__299keLO7vGoGbPf4rrBp_3a3BwrQxwSL0NEeU05WWm3-ui32q-tx_Z-n_urb2T8Ryq23o4y_915Ij742ylT-55e5_zybm4vrm1uv1R_Ukpji</recordid><startdate>199401</startdate><enddate>199401</enddate><creator>Shi, C. Z.</creator><creator>Collins, H. W.</creator><creator>Garside, W. T.</creator><creator>Buettger, C. W.</creator><creator>Matschinsky, F. M.</creator><creator>Heyner, S.</creator><general>Wiley Subscription Services, Inc., A Wiley Company</general><general>Wiley-Liss</general><scope>BSCLL</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>199401</creationdate><title>Protein databases for compacted eight-cell and blastocyst-stage mouse embryos</title><author>Shi, C. Z. ; Collins, H. W. ; Garside, W. T. ; Buettger, C. W. ; Matschinsky, F. M. ; Heyner, S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3886-cf4ac91b9f4e4ee0f0fbdf093284f649e5f767da87a2c1834eb97a5a25d423b23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Blastocyst - cytology</topic><topic>Blastocyst - metabolism</topic><topic>Blastocyst - physiology</topic><topic>Cell Division</topic><topic>Databases, Factual</topic><topic>Early stages. Segmentation. Gastrulation. Neurulation</topic><topic>Electrophoresis, Gel, Two-Dimensional</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Embryology: invertebrates and vertebrates. Teratology</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Male</topic><topic>Methionine - metabolism</topic><topic>Mice</topic><topic>Mice, Inbred Strains</topic><topic>Molecular Weight</topic><topic>PDQUEST</topic><topic>Preimplantation development</topic><topic>Protein Biosynthesis</topic><topic>Proteins - analysis</topic><topic>Proteins - isolation & purification</topic><topic>Two-dimensional gel electrophoresis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Shi, C. Z.</creatorcontrib><creatorcontrib>Collins, H. W.</creatorcontrib><creatorcontrib>Garside, W. T.</creatorcontrib><creatorcontrib>Buettger, C. W.</creatorcontrib><creatorcontrib>Matschinsky, F. 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Dev</addtitle><date>1994-01</date><risdate>1994</risdate><volume>37</volume><issue>1</issue><spage>34</spage><epage>47</epage><pages>34-47</pages><issn>1040-452X</issn><eissn>1098-2795</eissn><coden>MREDEE</coden><abstract>High‐resolution two‐dimensional sodium dodecyl sulfate‐polyacrylamide (2D‐SDS) gel electrophoresis combined with computerized analysis of gel images was used to construct and analyze protein databases for two stages of preimplantation mouse embryogenesis, the compacted eight‐cell stage and the fully expanded blastocyst stage. These stages were chosen for their ease in identification of multiple synchronous embryos. Synchronous cohorts of 30–50 embryos were labelled with L‐[35S]methionine for 2 hr. The embryos were then lysed in 30 μl hot SDS sample buffer, and the lysates were stored at −80°C until the gels were run. Five replicates were run for eight‐cell embryos, and four for blastocyst‐stage embryos. The samples were processed for 2D gel electrophoresis and fluorography; multiple exposures were made. Gel images were analyzed using the PDQUEST system, and databases were constructed. Analysis of the databases for both developmental stages showed high reproducibility of protein spots in multiple gel images. Of 1,674 total spots in eight‐cell embryo standards, >79% of spots had a percentage error (S.E.M./average) <50%, and >45% had a percentage error <30%. Similarly, of 1,653 total spots in blastocyst‐stage embryo standards, 74% of spots had a percentage error <50%, and approximately 47% of spots had a percentage error <30%. Forty‐three spots (approximately 3% of the total spots) were found to be detected only in the eight‐cell stage, while 75 spots were detected solely in the blastocyst stage. Sixty‐nine proteins showed a greater than threefold increase in isotope incorporation from the eight‐cell to the blastocyst stage, with a percentage error <50% in both the eight‐cell and the blastocyst stages. In contrast, 41 of the proteins showed a decrease during this period. Analysis of the protein databases described in this study has allowed us to document the overall quantitative changes in proteins from the compacted eight‐cell stage to the blastocyst stage of mouse preimplantation development. These databases provide a valuable tool for further detailed quantitative analysis of specific proteins associated with developmental events. In addition they will permit analysis of the effects of environmental factors, such as growth factors, on early embryo development. © 1994 Wiley‐Liss, Inc.</abstract><cop>Hoboken</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><pmid>8129929</pmid><doi>10.1002/mrd.1080370106</doi><tpages>14</tpages></addata></record> |
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| ispartof | Molecular reproduction and development, 1994-01, Vol.37 (1), p.34-47 |
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| subjects | Animals Biological and medical sciences Blastocyst - cytology Blastocyst - metabolism Blastocyst - physiology Cell Division Databases, Factual Early stages. Segmentation. Gastrulation. Neurulation Electrophoresis, Gel, Two-Dimensional Electrophoresis, Polyacrylamide Gel Embryology: invertebrates and vertebrates. Teratology Female Fundamental and applied biological sciences. Psychology Male Methionine - metabolism Mice Mice, Inbred Strains Molecular Weight PDQUEST Preimplantation development Protein Biosynthesis Proteins - analysis Proteins - isolation & purification Two-dimensional gel electrophoresis |
| title | Protein databases for compacted eight-cell and blastocyst-stage mouse embryos |
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