Visualization of primordial germ cells in the fertilized pelagic eggs of the barfin flounder Verasper moseri
Primordial germ cells (PGCs) appear during early embryogenesis and differentiate into gametes through oogenesis or spermatogenesis. Teleost PGCs can be visualized by injecting RNA transcribed from the fusion product of a fluorescent protein gene attached to the 3' untranslated region (3'UT...
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| Veröffentlicht in: | The International journal of developmental biology 2015, Vol.59 (10-12), p.465-470 |
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| container_issue | 10-12 |
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| container_title | The International journal of developmental biology |
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| creator | Goto, Rie Saito, Taiju Kawakami, Yutaka Kitauchi, Tomoe Takagi, Misae Todo, Takashi Arai, Katsutoshi Yamaha, Etsuro |
| description | Primordial germ cells (PGCs) appear during early embryogenesis and differentiate into gametes through oogenesis or spermatogenesis. Teleost PGCs can be visualized by injecting RNA transcribed from the fusion product of a fluorescent protein gene attached to the 3' untranslated region (3'UTR) of zebrafish nanos3 (zf-nos3). Although this method has been widely applied to teleost PGCs, the visualization of PGCs in pelagic species that have eggs with a hard chorion is more problematic due to the technical difficulty of microinjection into their eggs. In this study, we developed a reliable method for microinjection of fertilized eggs in a pelagic species, the barfin flounder. Using a microneedle with a constriction "brake", we were able to introduce gfp-nos3 3'UTR mRNA into embryos and to determine the origin and migration route of PGCs. We also isolated the barfin flounder nos3 (bf-nos3) gene to compare its 3'UTR sequence with that of zebrafish. The 3'UTR of the bf-nos3 sequence was longer than that of zf-nos3. However, PGCs were also visualized after injection of gfp-bf-nos3 3'UTR mRNA both in zebrafish and barfin flounder. These results suggest that the function of nos3 is conserved between these species regardless of the sequence differences. The method developed here for labeling PGCs with gfp-nos3 mRNA will provide a means to study PGC development in the embryos of a wide range of marine fish species. |
| doi_str_mv | 10.1387/ijdb.150008rg |
| format | Article |
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Teleost PGCs can be visualized by injecting RNA transcribed from the fusion product of a fluorescent protein gene attached to the 3' untranslated region (3'UTR) of zebrafish nanos3 (zf-nos3). Although this method has been widely applied to teleost PGCs, the visualization of PGCs in pelagic species that have eggs with a hard chorion is more problematic due to the technical difficulty of microinjection into their eggs. In this study, we developed a reliable method for microinjection of fertilized eggs in a pelagic species, the barfin flounder. Using a microneedle with a constriction "brake", we were able to introduce gfp-nos3 3'UTR mRNA into embryos and to determine the origin and migration route of PGCs. We also isolated the barfin flounder nos3 (bf-nos3) gene to compare its 3'UTR sequence with that of zebrafish. The 3'UTR of the bf-nos3 sequence was longer than that of zf-nos3. However, PGCs were also visualized after injection of gfp-bf-nos3 3'UTR mRNA both in zebrafish and barfin flounder. These results suggest that the function of nos3 is conserved between these species regardless of the sequence differences. The method developed here for labeling PGCs with gfp-nos3 mRNA will provide a means to study PGC development in the embryos of a wide range of marine fish species.</description><identifier>ISSN: 0214-6282</identifier><identifier>EISSN: 1696-3547</identifier><identifier>DOI: 10.1387/ijdb.150008rg</identifier><identifier>PMID: 26864487</identifier><language>eng</language><publisher>Spain</publisher><subject>3' Untranslated Regions - genetics ; Amino Acid Sequence ; Animals ; Danio rerio ; Embryo, Nonmammalian - cytology ; Embryo, Nonmammalian - metabolism ; Embryonic Development - physiology ; Flounder - embryology ; Freshwater ; Germ Cells - cytology ; Germ Cells - metabolism ; In Situ Hybridization ; Marine ; Molecular Sequence Data ; Pleuronectiformes ; Sequence Homology, Amino Acid ; Teleostei ; Verasper moseri ; Zebrafish - growth & development ; Zebrafish - metabolism ; Zebrafish Proteins - genetics ; Zebrafish Proteins - metabolism ; Zygote - cytology ; Zygote - metabolism</subject><ispartof>The International journal of developmental biology, 2015, Vol.59 (10-12), p.465-470</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c501t-876e621f4dacfbceacbe700661a3382cad744cdc34c0fade9aa00825b80bee943</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,4010,27900,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26864487$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Goto, Rie</creatorcontrib><creatorcontrib>Saito, Taiju</creatorcontrib><creatorcontrib>Kawakami, Yutaka</creatorcontrib><creatorcontrib>Kitauchi, Tomoe</creatorcontrib><creatorcontrib>Takagi, Misae</creatorcontrib><creatorcontrib>Todo, Takashi</creatorcontrib><creatorcontrib>Arai, Katsutoshi</creatorcontrib><creatorcontrib>Yamaha, Etsuro</creatorcontrib><title>Visualization of primordial germ cells in the fertilized pelagic eggs of the barfin flounder Verasper moseri</title><title>The International journal of developmental biology</title><addtitle>Int J Dev Biol</addtitle><description>Primordial germ cells (PGCs) appear during early embryogenesis and differentiate into gametes through oogenesis or spermatogenesis. 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However, PGCs were also visualized after injection of gfp-bf-nos3 3'UTR mRNA both in zebrafish and barfin flounder. These results suggest that the function of nos3 is conserved between these species regardless of the sequence differences. The method developed here for labeling PGCs with gfp-nos3 mRNA will provide a means to study PGC development in the embryos of a wide range of marine fish species.</description><subject>3' Untranslated Regions - genetics</subject><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Danio rerio</subject><subject>Embryo, Nonmammalian - cytology</subject><subject>Embryo, Nonmammalian - metabolism</subject><subject>Embryonic Development - physiology</subject><subject>Flounder - embryology</subject><subject>Freshwater</subject><subject>Germ Cells - cytology</subject><subject>Germ Cells - metabolism</subject><subject>In Situ Hybridization</subject><subject>Marine</subject><subject>Molecular Sequence Data</subject><subject>Pleuronectiformes</subject><subject>Sequence Homology, Amino Acid</subject><subject>Teleostei</subject><subject>Verasper moseri</subject><subject>Zebrafish - growth & development</subject><subject>Zebrafish - metabolism</subject><subject>Zebrafish Proteins - genetics</subject><subject>Zebrafish Proteins - metabolism</subject><subject>Zygote - cytology</subject><subject>Zygote - metabolism</subject><issn>0214-6282</issn><issn>1696-3547</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkb1PwzAQxS0EoqUwsiKPLAE7cWxnRBVfUiUW6Bo59jm4SuJgJwP89SRqy8x0d7rfnfTeQ-iakjuaSXHvdqa6ozkhRIb6BC0pL3iS5UycoiVJKUt4KtMFuohxR6aZSHGOFimXnDEplqjZujiqxv2owfkOe4v74FofjFMNriG0WEPTROw6PHwCthAGN9FgcA-Nqp3GUNdxvpvXlQp2Im3jx85AwFsIKvZT0_oIwV2iM6uaCFeHukIfT4_v65dk8_b8un7YJDondEik4MBTaplR2lYalK5AEMI5VVkmU62MYEwbnTFNrDJQKDWpT_NKkgqgYNkK3e7_9sF_jRCHsnVx1qE68GMsqRCcTwYQ-g-U5wUtiMwmNNmjOvgYA9hytkqF75KScs6inLMoj1lM_M3h9Vi1YP7oo_nZL76SiBU</recordid><startdate>2015</startdate><enddate>2015</enddate><creator>Goto, Rie</creator><creator>Saito, Taiju</creator><creator>Kawakami, Yutaka</creator><creator>Kitauchi, Tomoe</creator><creator>Takagi, Misae</creator><creator>Todo, Takashi</creator><creator>Arai, Katsutoshi</creator><creator>Yamaha, Etsuro</creator><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><scope>7TN</scope><scope>8FD</scope><scope>F1W</scope><scope>FR3</scope><scope>H95</scope><scope>L.G</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>2015</creationdate><title>Visualization of primordial germ cells in the fertilized pelagic eggs of the barfin flounder Verasper moseri</title><author>Goto, Rie ; 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Teleost PGCs can be visualized by injecting RNA transcribed from the fusion product of a fluorescent protein gene attached to the 3' untranslated region (3'UTR) of zebrafish nanos3 (zf-nos3). Although this method has been widely applied to teleost PGCs, the visualization of PGCs in pelagic species that have eggs with a hard chorion is more problematic due to the technical difficulty of microinjection into their eggs. In this study, we developed a reliable method for microinjection of fertilized eggs in a pelagic species, the barfin flounder. Using a microneedle with a constriction "brake", we were able to introduce gfp-nos3 3'UTR mRNA into embryos and to determine the origin and migration route of PGCs. We also isolated the barfin flounder nos3 (bf-nos3) gene to compare its 3'UTR sequence with that of zebrafish. The 3'UTR of the bf-nos3 sequence was longer than that of zf-nos3. 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| subjects | 3' Untranslated Regions - genetics Amino Acid Sequence Animals Danio rerio Embryo, Nonmammalian - cytology Embryo, Nonmammalian - metabolism Embryonic Development - physiology Flounder - embryology Freshwater Germ Cells - cytology Germ Cells - metabolism In Situ Hybridization Marine Molecular Sequence Data Pleuronectiformes Sequence Homology, Amino Acid Teleostei Verasper moseri Zebrafish - growth & development Zebrafish - metabolism Zebrafish Proteins - genetics Zebrafish Proteins - metabolism Zygote - cytology Zygote - metabolism |
| title | Visualization of primordial germ cells in the fertilized pelagic eggs of the barfin flounder Verasper moseri |
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