Effects of specific and prolonged expression of zebrafish growth factors, Fgf2 and Lif in primordial germ cells in vivo
► We discovered that nanos3 3′UTR prolonged PGC-specific protein expression up to 26days. ► Expression of Fgf2 in PGCs significantly increased PGC number at later developmental stages. ► Expression of Lif in PGCs resulted in a significant disruption of PGC migration. ► Lif illicited its effect on PG...
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| description | ► We discovered that nanos3 3′UTR prolonged PGC-specific protein expression up to 26days. ► Expression of Fgf2 in PGCs significantly increased PGC number at later developmental stages. ► Expression of Lif in PGCs resulted in a significant disruption of PGC migration. ► Lif illicited its effect on PGC migration through Lif receptor a. ► Our approach could be used to achieve prolonged PGC-specific expression of other proteins.
Primordial germ cells (PGCs), specified early in development, proliferate and migrate to the developing gonad before sexual differentiation occurs in the embryo and eventually give rise to spermatogonia or oogonia. In this study, we discovered that nanos3 3′UTR, a common method used to label PGCs, not only directed PGC-specific expression of DsRed but also prolonged this expression up to 26days post fertilization (dpf) when DsRed-nanos3 3′UTR hybrid mRNAs were introduced into 1- to 2-cell-stage embryos. As such, we employed this knowledge to express zebrafish leukemia inhibitory factor (Lif), basic fibroblast growth factor (Fgf2) and bone morphogenetic protein 4 (Bmp4) in the PGCs and evaluate their effects on PGC development in vivo for over a period of 3weeks. The results show that expression of Fgf2 significantly increased PGC number at 14- and 21-dpf while Bmp4 resulted in severe ventralization and death of the embryos by 3days. Expression of Lif resulted in a significant disruption of PGC migration. Mopholino knockdown experiments indicated that Lif illicited its effect on PGC migration through Lif receptor a (Lifra) but not Lifrb. The general approach described in this study could be used to achieve prolonged PGC-specific expression of other proteins to investigate their roles in germ cell and gonad development. The results also indicate that zebrafish PGCs have a mechanism to stabilize and prolong the expression of mRNA that carries nanos3 3′UTR. Understanding this mechanism may make it possible to achieve prolonged RNA expression in other cell types. |
| doi_str_mv | 10.1016/j.bbrc.2012.11.014 |
| format | Article |
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Primordial germ cells (PGCs), specified early in development, proliferate and migrate to the developing gonad before sexual differentiation occurs in the embryo and eventually give rise to spermatogonia or oogonia. In this study, we discovered that nanos3 3′UTR, a common method used to label PGCs, not only directed PGC-specific expression of DsRed but also prolonged this expression up to 26days post fertilization (dpf) when DsRed-nanos3 3′UTR hybrid mRNAs were introduced into 1- to 2-cell-stage embryos. As such, we employed this knowledge to express zebrafish leukemia inhibitory factor (Lif), basic fibroblast growth factor (Fgf2) and bone morphogenetic protein 4 (Bmp4) in the PGCs and evaluate their effects on PGC development in vivo for over a period of 3weeks. The results show that expression of Fgf2 significantly increased PGC number at 14- and 21-dpf while Bmp4 resulted in severe ventralization and death of the embryos by 3days. Expression of Lif resulted in a significant disruption of PGC migration. Mopholino knockdown experiments indicated that Lif illicited its effect on PGC migration through Lif receptor a (Lifra) but not Lifrb. The general approach described in this study could be used to achieve prolonged PGC-specific expression of other proteins to investigate their roles in germ cell and gonad development. The results also indicate that zebrafish PGCs have a mechanism to stabilize and prolong the expression of mRNA that carries nanos3 3′UTR. Understanding this mechanism may make it possible to achieve prolonged RNA expression in other cell types.</description><identifier>ISSN: 0006-291X</identifier><identifier>EISSN: 1090-2104</identifier><identifier>DOI: 10.1016/j.bbrc.2012.11.014</identifier><identifier>PMID: 23178298</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>3' untranslated regions ; 3' Untranslated Regions - genetics ; 60 APPLIED LIFE SCIENCES ; Animals ; Bone Morphogenetic Protein 4 - biosynthesis ; Bone Morphogenetic Protein 4 - genetics ; bone morphogenetic proteins ; Cell Count ; Cell Movement ; Danio rerio ; death ; early development ; EMBRYOS ; FERTILIZATION ; Fgf2 ; fibroblast growth factor 2 ; Fibroblast Growth Factor 2 - biosynthesis ; Fibroblast Growth Factor 2 - genetics ; FIBROBLASTS ; Freshwater ; gene expression ; Germ Cells - metabolism ; GONADS ; GROWTH FACTORS ; hybrids ; IN VIVO ; LEUKEMIA ; leukemia inhibitory factor ; Leukemia Inhibitory Factor - biosynthesis ; Leukemia Inhibitory Factor - genetics ; Leukemia Inhibitory Factor Receptor alpha Subunit - metabolism ; Lif ; MESSENGER-RNA ; nanos3 3′UTR ; OOGONIA ; PGC migration ; Primordial Germ Cells ; RECEPTORS ; RNA, Messenger - biosynthesis ; RNA-Binding Proteins ; SKELETON ; SPERMATOGONIA ; Zebrafish ; Zebrafish - genetics ; Zebrafish - growth & development ; Zebrafish Proteins - biosynthesis ; Zebrafish Proteins - genetics</subject><ispartof>Biochemical and biophysical research communications, 2013-01, Vol.430 (1), p.347-351</ispartof><rights>2012 Elsevier Inc.</rights><rights>Copyright © 2012 Elsevier Inc. All rights reserved.</rights><rights>2012 Elsevier Inc. All rights reserved. 2012</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c549t-893fc46524987d771b1b7ce0f630cab690ab9fa187542bac44c23c6a7cded7fc3</citedby><cites>FETCH-LOGICAL-c549t-893fc46524987d771b1b7ce0f630cab690ab9fa187542bac44c23c6a7cded7fc3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.bbrc.2012.11.014$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>230,314,776,780,881,3536,27903,27904,45974</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23178298$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://www.osti.gov/biblio/22210373$$D View this record in Osti.gov$$Hfree_for_read</backlink></links><search><creatorcontrib>Wong, Ten-Tsao</creatorcontrib><creatorcontrib>Collodi, Paul</creatorcontrib><title>Effects of specific and prolonged expression of zebrafish growth factors, Fgf2 and Lif in primordial germ cells in vivo</title><title>Biochemical and biophysical research communications</title><addtitle>Biochem Biophys Res Commun</addtitle><description>► We discovered that nanos3 3′UTR prolonged PGC-specific protein expression up to 26days. ► Expression of Fgf2 in PGCs significantly increased PGC number at later developmental stages. ► Expression of Lif in PGCs resulted in a significant disruption of PGC migration. ► Lif illicited its effect on PGC migration through Lif receptor a. ► Our approach could be used to achieve prolonged PGC-specific expression of other proteins.
Primordial germ cells (PGCs), specified early in development, proliferate and migrate to the developing gonad before sexual differentiation occurs in the embryo and eventually give rise to spermatogonia or oogonia. In this study, we discovered that nanos3 3′UTR, a common method used to label PGCs, not only directed PGC-specific expression of DsRed but also prolonged this expression up to 26days post fertilization (dpf) when DsRed-nanos3 3′UTR hybrid mRNAs were introduced into 1- to 2-cell-stage embryos. As such, we employed this knowledge to express zebrafish leukemia inhibitory factor (Lif), basic fibroblast growth factor (Fgf2) and bone morphogenetic protein 4 (Bmp4) in the PGCs and evaluate their effects on PGC development in vivo for over a period of 3weeks. The results show that expression of Fgf2 significantly increased PGC number at 14- and 21-dpf while Bmp4 resulted in severe ventralization and death of the embryos by 3days. Expression of Lif resulted in a significant disruption of PGC migration. Mopholino knockdown experiments indicated that Lif illicited its effect on PGC migration through Lif receptor a (Lifra) but not Lifrb. The general approach described in this study could be used to achieve prolonged PGC-specific expression of other proteins to investigate their roles in germ cell and gonad development. The results also indicate that zebrafish PGCs have a mechanism to stabilize and prolong the expression of mRNA that carries nanos3 3′UTR. Understanding this mechanism may make it possible to achieve prolonged RNA expression in other cell types.</description><subject>3' untranslated regions</subject><subject>3' Untranslated Regions - genetics</subject><subject>60 APPLIED LIFE SCIENCES</subject><subject>Animals</subject><subject>Bone Morphogenetic Protein 4 - biosynthesis</subject><subject>Bone Morphogenetic Protein 4 - genetics</subject><subject>bone morphogenetic proteins</subject><subject>Cell Count</subject><subject>Cell Movement</subject><subject>Danio rerio</subject><subject>death</subject><subject>early development</subject><subject>EMBRYOS</subject><subject>FERTILIZATION</subject><subject>Fgf2</subject><subject>fibroblast growth factor 2</subject><subject>Fibroblast Growth Factor 2 - biosynthesis</subject><subject>Fibroblast Growth Factor 2 - genetics</subject><subject>FIBROBLASTS</subject><subject>Freshwater</subject><subject>gene expression</subject><subject>Germ Cells - metabolism</subject><subject>GONADS</subject><subject>GROWTH FACTORS</subject><subject>hybrids</subject><subject>IN VIVO</subject><subject>LEUKEMIA</subject><subject>leukemia inhibitory factor</subject><subject>Leukemia Inhibitory Factor - biosynthesis</subject><subject>Leukemia Inhibitory Factor - genetics</subject><subject>Leukemia Inhibitory Factor Receptor alpha Subunit - metabolism</subject><subject>Lif</subject><subject>MESSENGER-RNA</subject><subject>nanos3 3′UTR</subject><subject>OOGONIA</subject><subject>PGC migration</subject><subject>Primordial Germ Cells</subject><subject>RECEPTORS</subject><subject>RNA, Messenger - biosynthesis</subject><subject>RNA-Binding Proteins</subject><subject>SKELETON</subject><subject>SPERMATOGONIA</subject><subject>Zebrafish</subject><subject>Zebrafish - genetics</subject><subject>Zebrafish - growth & development</subject><subject>Zebrafish Proteins - biosynthesis</subject><subject>Zebrafish Proteins - genetics</subject><issn>0006-291X</issn><issn>1090-2104</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkkFv1DAQhSMEokvhD3BAlrhwIMFjO3EiISRUtYC0EheQuFnOZJz1KhsvdnYL_HqSbqngApx8mG_ezDy_LHsKvAAO1att0bYRC8FBFAAFB3UvWwFveC6Aq_vZinNe5aKBL2fZo5S2nAOoqnmYnQkJuhZNvcquL50jnBILjqU9oXcemR07to9hCGNPHaNv-0gp-TAu0A9qo3U-bVgfw_W0Yc7iFGJ6ya56J25a194xP84Kfhdi5-3Aeoo7hjQMaSkc_TE8zh44OyR6cvueZ5-vLj9dvM_XH999uHi7zrFUzZTXjXSoqlKoptad1tBCq5G4qyRH21YNt23jLNS6VKK1qBQKiZXV2FGnHcrz7M1Jd39od9QhjVO0g1l2s_G7CdabPyuj35g-HI0s1TyzngWenwRCmrxJ6CfCDYZxnF0zQsxOSy1n6sXtmBi-HihNZufTcrAdKRySAV3WQtYVlP-BCq6kUrL5Nyq0LHlV3qwpTijGkFIkd3cicLOkxWzNkhazpMUAmDktc9Oz3825a_kVjxl4fQJo_qKjp7gYQCNS5-Nyfxf83_R_ArrI0Zo</recordid><startdate>20130104</startdate><enddate>20130104</enddate><creator>Wong, Ten-Tsao</creator><creator>Collodi, Paul</creator><general>Elsevier Inc</general><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>7S9</scope><scope>L.6</scope><scope>F1W</scope><scope>H95</scope><scope>L.G</scope><scope>OTOTI</scope><scope>5PM</scope></search><sort><creationdate>20130104</creationdate><title>Effects of specific and prolonged expression of zebrafish growth factors, Fgf2 and Lif in primordial germ cells in vivo</title><author>Wong, Ten-Tsao ; Collodi, Paul</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c549t-893fc46524987d771b1b7ce0f630cab690ab9fa187542bac44c23c6a7cded7fc3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>3' untranslated regions</topic><topic>3' Untranslated Regions - genetics</topic><topic>60 APPLIED LIFE SCIENCES</topic><topic>Animals</topic><topic>Bone Morphogenetic Protein 4 - biosynthesis</topic><topic>Bone Morphogenetic Protein 4 - genetics</topic><topic>bone morphogenetic proteins</topic><topic>Cell Count</topic><topic>Cell Movement</topic><topic>Danio rerio</topic><topic>death</topic><topic>early development</topic><topic>EMBRYOS</topic><topic>FERTILIZATION</topic><topic>Fgf2</topic><topic>fibroblast growth factor 2</topic><topic>Fibroblast Growth Factor 2 - biosynthesis</topic><topic>Fibroblast Growth Factor 2 - genetics</topic><topic>FIBROBLASTS</topic><topic>Freshwater</topic><topic>gene expression</topic><topic>Germ Cells - metabolism</topic><topic>GONADS</topic><topic>GROWTH FACTORS</topic><topic>hybrids</topic><topic>IN VIVO</topic><topic>LEUKEMIA</topic><topic>leukemia inhibitory factor</topic><topic>Leukemia Inhibitory Factor - biosynthesis</topic><topic>Leukemia Inhibitory Factor - genetics</topic><topic>Leukemia Inhibitory Factor Receptor alpha Subunit - metabolism</topic><topic>Lif</topic><topic>MESSENGER-RNA</topic><topic>nanos3 3′UTR</topic><topic>OOGONIA</topic><topic>PGC migration</topic><topic>Primordial Germ Cells</topic><topic>RECEPTORS</topic><topic>RNA, Messenger - biosynthesis</topic><topic>RNA-Binding Proteins</topic><topic>SKELETON</topic><topic>SPERMATOGONIA</topic><topic>Zebrafish</topic><topic>Zebrafish - genetics</topic><topic>Zebrafish - growth & development</topic><topic>Zebrafish Proteins - biosynthesis</topic><topic>Zebrafish Proteins - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wong, Ten-Tsao</creatorcontrib><creatorcontrib>Collodi, Paul</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>AGRICOLA</collection><collection>AGRICOLA - Academic</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 1: Biological Sciences & Living Resources</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><collection>OSTI.GOV</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Biochemical and biophysical research communications</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wong, Ten-Tsao</au><au>Collodi, Paul</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Effects of specific and prolonged expression of zebrafish growth factors, Fgf2 and Lif in primordial germ cells in vivo</atitle><jtitle>Biochemical and biophysical research communications</jtitle><addtitle>Biochem Biophys Res Commun</addtitle><date>2013-01-04</date><risdate>2013</risdate><volume>430</volume><issue>1</issue><spage>347</spage><epage>351</epage><pages>347-351</pages><issn>0006-291X</issn><eissn>1090-2104</eissn><abstract>► We discovered that nanos3 3′UTR prolonged PGC-specific protein expression up to 26days. ► Expression of Fgf2 in PGCs significantly increased PGC number at later developmental stages. ► Expression of Lif in PGCs resulted in a significant disruption of PGC migration. ► Lif illicited its effect on PGC migration through Lif receptor a. ► Our approach could be used to achieve prolonged PGC-specific expression of other proteins.
Primordial germ cells (PGCs), specified early in development, proliferate and migrate to the developing gonad before sexual differentiation occurs in the embryo and eventually give rise to spermatogonia or oogonia. In this study, we discovered that nanos3 3′UTR, a common method used to label PGCs, not only directed PGC-specific expression of DsRed but also prolonged this expression up to 26days post fertilization (dpf) when DsRed-nanos3 3′UTR hybrid mRNAs were introduced into 1- to 2-cell-stage embryos. As such, we employed this knowledge to express zebrafish leukemia inhibitory factor (Lif), basic fibroblast growth factor (Fgf2) and bone morphogenetic protein 4 (Bmp4) in the PGCs and evaluate their effects on PGC development in vivo for over a period of 3weeks. The results show that expression of Fgf2 significantly increased PGC number at 14- and 21-dpf while Bmp4 resulted in severe ventralization and death of the embryos by 3days. Expression of Lif resulted in a significant disruption of PGC migration. Mopholino knockdown experiments indicated that Lif illicited its effect on PGC migration through Lif receptor a (Lifra) but not Lifrb. The general approach described in this study could be used to achieve prolonged PGC-specific expression of other proteins to investigate their roles in germ cell and gonad development. The results also indicate that zebrafish PGCs have a mechanism to stabilize and prolong the expression of mRNA that carries nanos3 3′UTR. Understanding this mechanism may make it possible to achieve prolonged RNA expression in other cell types.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>23178298</pmid><doi>10.1016/j.bbrc.2012.11.014</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Biochemical and biophysical research communications, 2013-01, Vol.430 (1), p.347-351 |
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| subjects | 3' untranslated regions 3' Untranslated Regions - genetics 60 APPLIED LIFE SCIENCES Animals Bone Morphogenetic Protein 4 - biosynthesis Bone Morphogenetic Protein 4 - genetics bone morphogenetic proteins Cell Count Cell Movement Danio rerio death early development EMBRYOS FERTILIZATION Fgf2 fibroblast growth factor 2 Fibroblast Growth Factor 2 - biosynthesis Fibroblast Growth Factor 2 - genetics FIBROBLASTS Freshwater gene expression Germ Cells - metabolism GONADS GROWTH FACTORS hybrids IN VIVO LEUKEMIA leukemia inhibitory factor Leukemia Inhibitory Factor - biosynthesis Leukemia Inhibitory Factor - genetics Leukemia Inhibitory Factor Receptor alpha Subunit - metabolism Lif MESSENGER-RNA nanos3 3′UTR OOGONIA PGC migration Primordial Germ Cells RECEPTORS RNA, Messenger - biosynthesis RNA-Binding Proteins SKELETON SPERMATOGONIA Zebrafish Zebrafish - genetics Zebrafish - growth & development Zebrafish Proteins - biosynthesis Zebrafish Proteins - genetics |
| title | Effects of specific and prolonged expression of zebrafish growth factors, Fgf2 and Lif in primordial germ cells in vivo |
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