Novel technique for visualizing primordial germ cells in sturgeons (Acipenser ruthenus, A. gueldenstaedtii, A. baerii, and Huso huso).
Identifieur interne : 000641 ( Ncbi/Merge ); précédent : 000640; suivant : 000642Novel technique for visualizing primordial germ cells in sturgeons (Acipenser ruthenus, A. gueldenstaedtii, A. baerii, and Huso huso).
Auteurs : Taiju Saito [Oman] ; Martin Psenicka [République tchèque]Source :
- Biology of reproduction [ 1529-7268 ] ; 2015.
English descriptors
- KwdEn :
- Animals, Cell Movement, Dextrans, Embryo, Nonmammalian (ultrastructure), Embryonic Development (physiology), Female, Fishes (physiology), Fluorescein-5-isothiocyanate (analogs & derivatives), Fluorescent Dyes, Germ Cells (ultrastructure), Male, Microinjections, Molecular Weight, Sperm Tail (ultrastructure).
- MESH :
- chemical , analogs & derivatives : Fluorescein-5-isothiocyanate.
- chemical : Dextrans, Fluorescent Dyes.
- physiology : Embryonic Development, Fishes.
- ultrastructure : Embryo, Nonmammalian, Germ Cells, Sperm Tail.
- Animals, Cell Movement, Female, Male, Microinjections, Molecular Weight.
Abstract
Primordial germ cells (PGCs) are the origin of all germ cells in developing embryos. In the sturgeon embryo, PGCs develop from the vegetal hemisphere, which mainly acts as an extraembryonic source of nutrition. Current methods for studying sturgeon PGCs require either killing the fish or using costly and time-consuming histological procedures. Here, we demonstrate that visualization of sterlet (Acipenser ruthenus>) PGCs in vivo is feasible by simply labeling the vegetal hemisphere with fluorescein isothiocyanate (FITC)-dextran. We injected FITC-dextrans, with molecular weights varying between 10 000 and 2 000 000, into the vegetal pole of 1- to 4-cell stage embryos. At the neurula to tail-bud developmental stages, FITC-positive PGC-like cells appeared ventrally around the developing tail bud in the experimental group that received a high-molecular-weight FITC-dextran. The highest average number of FITC-positive PGC-like cells was observed in embryos injected with FITC-dextran having a molecular weight of 500 000 (FD-500). The pattern of migration of the labeled cells was identical to that of PGCs, clearly indicating that the FITC-positive PGC-like cells were PGCs. Labeled vegetal cells, except for the PGCs, were digested and excreted before the embryos starting feeding. FITC-labeled PGCs were observed in the developing gonads of fish for at least 3 mo after injection. We also found that FD-500 could be used to visualize PGCs in other sturgeon species. To the best of our knowledge, this report is the first to demonstrate in any animal species that PGCs can be visualized in vivo for a long period by the injection of a simple reagent.
DOI: 10.1095/biolreprod.115.128314
PubMed: 26134864
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Novel technique for visualizing primordial germ cells in sturgeons (Acipenser ruthenus, A. gueldenstaedtii, A. baerii, and Huso huso).</title><author><name sortKey="Saito, Taiju" sort="Saito, Taiju" uniqKey="Saito T" first="Taiju" last="Saito">Taiju Saito</name><affiliation wicri:level="1"><nlm:affiliation>Laboratory of Germ Cells, Research Institute of Fish Culture and Hydrobiology, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, University of South Bohemia in Ceske Budejovice, Vodnany, Czech Republic taiju76@gmail.com.</nlm:affiliation><country wicri:rule="url">Oman</country><wicri:regionArea>Laboratory of Germ Cells, Research Institute of Fish Culture and Hydrobiology, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, University of South Bohemia in Ceske Budejovice, Vodnany</wicri:regionArea><wicri:noRegion>Vodnany</wicri:noRegion></affiliation></author><author><name sortKey="Psenicka, Martin" sort="Psenicka, Martin" uniqKey="Psenicka M" first="Martin" last="Psenicka">Martin Psenicka</name><affiliation wicri:level="1"><nlm:affiliation>Laboratory of Germ Cells, Research Institute of Fish Culture and Hydrobiology, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, University of South Bohemia in Ceske Budejovice, Vodnany, Czech Republic.</nlm:affiliation><country xml:lang="fr">République tchèque</country><wicri:regionArea>Laboratory of Germ Cells, Research Institute of Fish Culture and Hydrobiology, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, University of South Bohemia in Ceske Budejovice, 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huso).</title><author><name sortKey="Saito, Taiju" sort="Saito, Taiju" uniqKey="Saito T" first="Taiju" last="Saito">Taiju Saito</name><affiliation wicri:level="1"><nlm:affiliation>Laboratory of Germ Cells, Research Institute of Fish Culture and Hydrobiology, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, University of South Bohemia in Ceske Budejovice, Vodnany, Czech Republic taiju76@gmail.com.</nlm:affiliation><country wicri:rule="url">Oman</country><wicri:regionArea>Laboratory of Germ Cells, Research Institute of Fish Culture and Hydrobiology, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, University of South Bohemia in Ceske Budejovice, Vodnany</wicri:regionArea><wicri:noRegion>Vodnany</wicri:noRegion></affiliation></author><author><name sortKey="Psenicka, Martin" sort="Psenicka, Martin" uniqKey="Psenicka M" first="Martin" last="Psenicka">Martin Psenicka</name><affiliation wicri:level="1"><nlm:affiliation>Laboratory of Germ Cells, Research Institute of Fish Culture and Hydrobiology, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, University of South Bohemia in Ceske Budejovice, Vodnany, Czech Republic.</nlm:affiliation><country xml:lang="fr">République tchèque</country><wicri:regionArea>Laboratory of Germ Cells, Research Institute of Fish Culture and Hydrobiology, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, University of South Bohemia in Ceske Budejovice, Vodnany</wicri:regionArea><wicri:noRegion>Vodnany</wicri:noRegion></affiliation></author></analytic><series><title level="j">Biology of reproduction</title><idno type="eISSN">1529-7268</idno><imprint><date when="2015" type="published">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Cell Movement</term><term>Dextrans</term><term>Embryo, Nonmammalian (ultrastructure)</term><term>Embryonic Development (physiology)</term><term>Female</term><term>Fishes (physiology)</term><term>Fluorescein-5-isothiocyanate (analogs & derivatives)</term><term>Fluorescent Dyes</term><term>Germ Cells (ultrastructure)</term><term>Male</term><term>Microinjections</term><term>Molecular Weight</term><term>Sperm Tail (ultrastructure)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analogs & derivatives" xml:lang="en"><term>Fluorescein-5-isothiocyanate</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Dextrans</term><term>Fluorescent Dyes</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Embryonic Development</term><term>Fishes</term></keywords><keywords scheme="MESH" qualifier="ultrastructure" xml:lang="en"><term>Embryo, Nonmammalian</term><term>Germ Cells</term><term>Sperm Tail</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Cell Movement</term><term>Female</term><term>Male</term><term>Microinjections</term><term>Molecular Weight</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Primordial germ cells (PGCs) are the origin of all germ cells in developing embryos. In the sturgeon embryo, PGCs develop from the vegetal hemisphere, which mainly acts as an extraembryonic source of nutrition. Current methods for studying sturgeon PGCs require either killing the fish or using costly and time-consuming histological procedures. Here, we demonstrate that visualization of sterlet (Acipenser ruthenus>) PGCs in vivo is feasible by simply labeling the vegetal hemisphere with fluorescein isothiocyanate (FITC)-dextran. We injected FITC-dextrans, with molecular weights varying between 10 000 and 2 000 000, into the vegetal pole of 1- to 4-cell stage embryos. At the neurula to tail-bud developmental stages, FITC-positive PGC-like cells appeared ventrally around the developing tail bud in the experimental group that received a high-molecular-weight FITC-dextran. The highest average number of FITC-positive PGC-like cells was observed in embryos injected with FITC-dextran having a molecular weight of 500 000 (FD-500). The pattern of migration of the labeled cells was identical to that of PGCs, clearly indicating that the FITC-positive PGC-like cells were PGCs. Labeled vegetal cells, except for the PGCs, were digested and excreted before the embryos starting feeding. FITC-labeled PGCs were observed in the developing gonads of fish for at least 3 mo after injection. We also found that FD-500 could be used to visualize PGCs in other sturgeon species. To the best of our knowledge, this report is the first to demonstrate in any animal species that PGCs can be visualized in vivo for a long period by the injection of a simple reagent.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">26134864</PMID><DateCreated><Year>2015</Year><Month>10</Month><Day>23</Day></DateCreated><DateCompleted><Year>2016</Year><Month>07</Month><Day>14</Day></DateCompleted><DateRevised><Year>2016</Year><Month>11</Month><Day>25</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1529-7268</ISSN><JournalIssue CitedMedium="Internet"><Volume>93</Volume><Issue>4</Issue><PubDate><Year>2015</Year><Month>Oct</Month></PubDate></JournalIssue><Title>Biology of reproduction</Title><ISOAbbreviation>Biol. Reprod.</ISOAbbreviation></Journal><ArticleTitle>Novel technique for visualizing primordial germ cells in sturgeons (Acipenser ruthenus, A. gueldenstaedtii, A. baerii, and Huso huso).</ArticleTitle><Pagination><MedlinePgn>96</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1095/biolreprod.115.128314</ELocationID><Abstract><AbstractText>Primordial germ cells (PGCs) are the origin of all germ cells in developing embryos. In the sturgeon embryo, PGCs develop from the vegetal hemisphere, which mainly acts as an extraembryonic source of nutrition. Current methods for studying sturgeon PGCs require either killing the fish or using costly and time-consuming histological procedures. Here, we demonstrate that visualization of sterlet (Acipenser ruthenus>) PGCs in vivo is feasible by simply labeling the vegetal hemisphere with fluorescein isothiocyanate (FITC)-dextran. We injected FITC-dextrans, with molecular weights varying between 10 000 and 2 000 000, into the vegetal pole of 1- to 4-cell stage embryos. At the neurula to tail-bud developmental stages, FITC-positive PGC-like cells appeared ventrally around the developing tail bud in the experimental group that received a high-molecular-weight FITC-dextran. The highest average number of FITC-positive PGC-like cells was observed in embryos injected with FITC-dextran having a molecular weight of 500 000 (FD-500). The pattern of migration of the labeled cells was identical to that of PGCs, clearly indicating that the FITC-positive PGC-like cells were PGCs. Labeled vegetal cells, except for the PGCs, were digested and excreted before the embryos starting feeding. FITC-labeled PGCs were observed in the developing gonads of fish for at least 3 mo after injection. We also found that FD-500 could be used to visualize PGCs in other sturgeon species. To the best of our knowledge, this report is the first to demonstrate in any animal species that PGCs can be visualized in vivo for a long period by the injection of a simple reagent.</AbstractText><CopyrightInformation>© 2015 by the Society for the Study of Reproduction, Inc.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Saito</LastName><ForeName>Taiju</ForeName><Initials>T</Initials><AffiliationInfo><Affiliation>Laboratory of Germ Cells, Research Institute of Fish Culture and Hydrobiology, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, University of South Bohemia in Ceske Budejovice, Vodnany, Czech Republic taiju76@gmail.com.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Psenicka</LastName><ForeName>Martin</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Laboratory of Germ Cells, Research Institute of Fish Culture and Hydrobiology, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, University of South Bohemia in Ceske Budejovice, Vodnany, Czech Republic.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2015</Year><Month>07</Month><Day>01</Day></ArticleDate></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Biol Reprod</MedlineTA><NlmUniqueID>0207224</NlmUniqueID><ISSNLinking>0006-3363</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D003911">Dextrans</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D005456">Fluorescent 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MajorTopicYN="N">Microinjections</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008970" MajorTopicYN="N">Molecular Weight</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D013082" MajorTopicYN="N">Sperm Tail</DescriptorName><QualifierName UI="Q000648" MajorTopicYN="N">ultrastructure</QualifierName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">developmental biology</Keyword><Keyword MajorTopicYN="N">early development</Keyword><Keyword MajorTopicYN="N">fish reproduction</Keyword><Keyword MajorTopicYN="N">gonad development</Keyword><Keyword MajorTopicYN="N">primordial germ cells</Keyword><Keyword MajorTopicYN="N">sturgeon</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2015</Year><Month>01</Month><Day>16</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2015</Year><Month>06</Month><Day>03</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2015</Year><Month>7</Month><Day>3</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2015</Year><Month>7</Month><Day>3</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2016</Year><Month>7</Month><Day>15</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">26134864</ArticleId><ArticleId IdType="pii">biolreprod.115.128314</ArticleId><ArticleId IdType="doi">10.1095/biolreprod.115.128314</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Oman</li><li>République tchèque</li></country></list><tree><country name="Oman"><noRegion><name sortKey="Saito, Taiju" sort="Saito, Taiju" uniqKey="Saito T" first="Taiju" last="Saito">Taiju Saito</name></noRegion></country><country name="République tchèque"><noRegion><name sortKey="Psenicka, Martin" sort="Psenicka, Martin" uniqKey="Psenicka M" first="Martin" last="Psenicka">Martin Psenicka</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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