Nucleolar Changes in Bovine Nucleotransferred Embryos
This study focused on nucleolar changes in bovine embryos reconstructed from enucleated mature oocytes fused with blastomeres of morulae or with cultured, serum unstarved bovine fetal skin fibroblasts (embryonic vs. somatic cloning). The nucleotransferred (NT) embryos were collected and fixed at tim...
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| Veröffentlicht in: | Biology of reproduction 2002-02, Vol.66 (2), p.534-543 |
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| creator | BARAN, V VIGNON, X LEBOURHIS, D RENARD, J. P FLECHON, J. E |
| description | This study focused on nucleolar changes in bovine embryos reconstructed from enucleated mature oocytes fused with blastomeres
of morulae or with cultured, serum unstarved bovine fetal skin fibroblasts (embryonic vs. somatic cloning). The nucleotransferred
(NT) embryos were collected and fixed at time intervals of 1â2 h (early 1-cell stage), 10â15 h (late 1-cell stage), 22â24
h (2-cell stage), 37â38 h (4-cell stage), 40â41 h (early 8-cell stage), 47â48 h (late 8-cell stage), and 55 h (16-cell stage)
after fusion. Immunocytochemistry by light and electron microscopy was used for structure-function characterization of nucleolar
components. Antibodies against RNA, protein B23, protein C23, and fibrillarin were applied. In addition, DNA was localized
by the terminal deoxynucleotidyl transferase (TdT) technique, and the functional organization of chromatin was determined
with the nick-translation immunogold approach. The results show that fully reticulated (active) nucleoli observed in donor
cells immediately before fusion as well as in the early 1-cell stage after fusion were progressively transformed into nucleolar
bodies displaying decreasing numbers of vacuoles from the 2- to 4-cell stage in both types of reconstructed embryos. At the
late 8-cell stage, morphological signs of resuming nucleolar activity were detected. Numerous new small vacuoles appeared,
and chromatin blocks reassociated with the nucleolar body. During this period, nick-translation technique revealed numerous
active DNA sites in the periphery of chromatin blocks associated with the nucleolar body. Fully reticulated nucleoli were
again observed as early as the 16-cell stage of embryonic cloned embryos. In comparison, the embryos obtained by fetal cloning
displayed a lower tendency to develop, mainly during the first cell cycle and during the period of presumed reactivation.
Correlatively, the changes in nucleolar morphology (desegregation and rebuilding) were at least delayed in many somatic NT
embryos in comparison with the embryonic NT group. It is concluded that complete reprogramming of rRNA gene expression is
part of the general nuclear reprogramming necessary for development after NT. |
| doi_str_mv | 10.1095/biolreprod66.2.534 |
| format | Article |
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of morulae or with cultured, serum unstarved bovine fetal skin fibroblasts (embryonic vs. somatic cloning). The nucleotransferred
(NT) embryos were collected and fixed at time intervals of 1â2 h (early 1-cell stage), 10â15 h (late 1-cell stage), 22â24
h (2-cell stage), 37â38 h (4-cell stage), 40â41 h (early 8-cell stage), 47â48 h (late 8-cell stage), and 55 h (16-cell stage)
after fusion. Immunocytochemistry by light and electron microscopy was used for structure-function characterization of nucleolar
components. Antibodies against RNA, protein B23, protein C23, and fibrillarin were applied. In addition, DNA was localized
by the terminal deoxynucleotidyl transferase (TdT) technique, and the functional organization of chromatin was determined
with the nick-translation immunogold approach. The results show that fully reticulated (active) nucleoli observed in donor
cells immediately before fusion as well as in the early 1-cell stage after fusion were progressively transformed into nucleolar
bodies displaying decreasing numbers of vacuoles from the 2- to 4-cell stage in both types of reconstructed embryos. At the
late 8-cell stage, morphological signs of resuming nucleolar activity were detected. Numerous new small vacuoles appeared,
and chromatin blocks reassociated with the nucleolar body. During this period, nick-translation technique revealed numerous
active DNA sites in the periphery of chromatin blocks associated with the nucleolar body. Fully reticulated nucleoli were
again observed as early as the 16-cell stage of embryonic cloned embryos. In comparison, the embryos obtained by fetal cloning
displayed a lower tendency to develop, mainly during the first cell cycle and during the period of presumed reactivation.
Correlatively, the changes in nucleolar morphology (desegregation and rebuilding) were at least delayed in many somatic NT
embryos in comparison with the embryonic NT group. It is concluded that complete reprogramming of rRNA gene expression is
part of the general nuclear reprogramming necessary for development after NT.</description><identifier>ISSN: 0006-3363</identifier><identifier>EISSN: 1529-7268</identifier><identifier>DOI: 10.1095/biolreprod66.2.534</identifier><identifier>PMID: 11804972</identifier><identifier>CODEN: BIREBV</identifier><language>eng</language><publisher>Madison, WI: Society for the Study of Reproduction</publisher><subject>Animals ; Antibiotics, Antineoplastic - pharmacology ; Antibodies, Monoclonal - immunology ; Biological and medical sciences ; Cattle ; Cell Fusion ; Cell Nucleolus - genetics ; Cell Nucleolus - metabolism ; Cell Nucleolus - ultrastructure ; Cloning, Organism ; Dactinomycin - pharmacology ; DNA - metabolism ; Embryo Transfer ; Embryonic and Fetal Development - physiology ; Female ; Fibroblasts - physiology ; Fluorescent Antibody Technique ; Gene Expression Regulation, Developmental - genetics ; Gene Expression Regulation, Developmental - physiology ; In Situ Nick-End Labeling ; Life Sciences ; Microscopy, Confocal ; Microscopy, Electron ; Microscopy, Fluorescence ; Oocytes - physiology ; Pregnancy</subject><ispartof>Biology of reproduction, 2002-02, Vol.66 (2), p.534-543</ispartof><rights>2002 INIST-CNRS</rights><rights>Distributed under a Creative Commons Attribution 4.0 International License</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,776,780,881,27903,27904</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=13447303$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11804972$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://hal.inrae.fr/hal-02671371$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>BARAN, V</creatorcontrib><creatorcontrib>VIGNON, X</creatorcontrib><creatorcontrib>LEBOURHIS, D</creatorcontrib><creatorcontrib>RENARD, J. P</creatorcontrib><creatorcontrib>FLECHON, J. E</creatorcontrib><title>Nucleolar Changes in Bovine Nucleotransferred Embryos</title><title>Biology of reproduction</title><addtitle>Biol Reprod</addtitle><description>This study focused on nucleolar changes in bovine embryos reconstructed from enucleated mature oocytes fused with blastomeres
of morulae or with cultured, serum unstarved bovine fetal skin fibroblasts (embryonic vs. somatic cloning). The nucleotransferred
(NT) embryos were collected and fixed at time intervals of 1â2 h (early 1-cell stage), 10â15 h (late 1-cell stage), 22â24
h (2-cell stage), 37â38 h (4-cell stage), 40â41 h (early 8-cell stage), 47â48 h (late 8-cell stage), and 55 h (16-cell stage)
after fusion. Immunocytochemistry by light and electron microscopy was used for structure-function characterization of nucleolar
components. Antibodies against RNA, protein B23, protein C23, and fibrillarin were applied. In addition, DNA was localized
by the terminal deoxynucleotidyl transferase (TdT) technique, and the functional organization of chromatin was determined
with the nick-translation immunogold approach. The results show that fully reticulated (active) nucleoli observed in donor
cells immediately before fusion as well as in the early 1-cell stage after fusion were progressively transformed into nucleolar
bodies displaying decreasing numbers of vacuoles from the 2- to 4-cell stage in both types of reconstructed embryos. At the
late 8-cell stage, morphological signs of resuming nucleolar activity were detected. Numerous new small vacuoles appeared,
and chromatin blocks reassociated with the nucleolar body. During this period, nick-translation technique revealed numerous
active DNA sites in the periphery of chromatin blocks associated with the nucleolar body. Fully reticulated nucleoli were
again observed as early as the 16-cell stage of embryonic cloned embryos. In comparison, the embryos obtained by fetal cloning
displayed a lower tendency to develop, mainly during the first cell cycle and during the period of presumed reactivation.
Correlatively, the changes in nucleolar morphology (desegregation and rebuilding) were at least delayed in many somatic NT
embryos in comparison with the embryonic NT group. It is concluded that complete reprogramming of rRNA gene expression is
part of the general nuclear reprogramming necessary for development after NT.</description><subject>Animals</subject><subject>Antibiotics, Antineoplastic - pharmacology</subject><subject>Antibodies, Monoclonal - immunology</subject><subject>Biological and medical sciences</subject><subject>Cattle</subject><subject>Cell Fusion</subject><subject>Cell Nucleolus - genetics</subject><subject>Cell Nucleolus - metabolism</subject><subject>Cell Nucleolus - ultrastructure</subject><subject>Cloning, Organism</subject><subject>Dactinomycin - pharmacology</subject><subject>DNA - metabolism</subject><subject>Embryo Transfer</subject><subject>Embryonic and Fetal Development - physiology</subject><subject>Female</subject><subject>Fibroblasts - physiology</subject><subject>Fluorescent Antibody Technique</subject><subject>Gene Expression Regulation, Developmental - genetics</subject><subject>Gene Expression Regulation, Developmental - physiology</subject><subject>In Situ Nick-End Labeling</subject><subject>Life Sciences</subject><subject>Microscopy, Confocal</subject><subject>Microscopy, Electron</subject><subject>Microscopy, Fluorescence</subject><subject>Oocytes - physiology</subject><subject>Pregnancy</subject><issn>0006-3363</issn><issn>1529-7268</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2002</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpF0U9PwjAYBvDGaATRL-DB7KKJh2H_dz0iQTEhetFzU7qO1XQbtoyFb-8IKKe-6fvLk_QpALcIjhGU7GnpGh_sOjQ552M8ZoSegSFiWKYC8-wcDCGEPCWEkwG4ivEbQkQJJpdggFAGqRR4CNh7a7xtvA7JtNT1ysbE1clzs3W1TQ67TdB1LGwINk9m1TLsmngNLgrto705niPw9TL7nM7Txcfr23SySEsC0SYthNZSZsYYzWCBrECZoJzmguc0E8vCCEMlhzkimhmeM4i0pNRyJPaDhGQEHg-5pfZqHVylw0412qn5ZKH2dxBzgYhAW9Tbh4PtC_lpbdyoykVjvde1bdqoBKIwYxL38O4I22Vl8__cv1J6cH8EOhrti_79xsWTI5QKAsnJlW5Vdi5YFSvtfR9LVNd1nCus-k8hv0q4fbQ</recordid><startdate>20020201</startdate><enddate>20020201</enddate><creator>BARAN, V</creator><creator>VIGNON, X</creator><creator>LEBOURHIS, D</creator><creator>RENARD, J. P</creator><creator>FLECHON, J. E</creator><general>Society for the Study of Reproduction</general><general>Society for the Study of Reproduction - Oxford Academic</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope><scope>1XC</scope></search><sort><creationdate>20020201</creationdate><title>Nucleolar Changes in Bovine Nucleotransferred Embryos</title><author>BARAN, V ; VIGNON, X ; LEBOURHIS, D ; RENARD, J. P ; FLECHON, J. E</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-h301t-f7aa998ccca50f1e7187464d76d487bfc7c4960d13a5c6d501a944e6171a94903</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2002</creationdate><topic>Animals</topic><topic>Antibiotics, Antineoplastic - pharmacology</topic><topic>Antibodies, Monoclonal - immunology</topic><topic>Biological and medical sciences</topic><topic>Cattle</topic><topic>Cell Fusion</topic><topic>Cell Nucleolus - genetics</topic><topic>Cell Nucleolus - metabolism</topic><topic>Cell Nucleolus - ultrastructure</topic><topic>Cloning, Organism</topic><topic>Dactinomycin - pharmacology</topic><topic>DNA - metabolism</topic><topic>Embryo Transfer</topic><topic>Embryonic and Fetal Development - physiology</topic><topic>Female</topic><topic>Fibroblasts - physiology</topic><topic>Fluorescent Antibody Technique</topic><topic>Gene Expression Regulation, Developmental - genetics</topic><topic>Gene Expression Regulation, Developmental - physiology</topic><topic>In Situ Nick-End Labeling</topic><topic>Life Sciences</topic><topic>Microscopy, Confocal</topic><topic>Microscopy, Electron</topic><topic>Microscopy, Fluorescence</topic><topic>Oocytes - physiology</topic><topic>Pregnancy</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>BARAN, V</creatorcontrib><creatorcontrib>VIGNON, X</creatorcontrib><creatorcontrib>LEBOURHIS, D</creatorcontrib><creatorcontrib>RENARD, J. P</creatorcontrib><creatorcontrib>FLECHON, J. E</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><collection>Hyper Article en Ligne (HAL)</collection><jtitle>Biology of reproduction</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>BARAN, V</au><au>VIGNON, X</au><au>LEBOURHIS, D</au><au>RENARD, J. P</au><au>FLECHON, J. E</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Nucleolar Changes in Bovine Nucleotransferred Embryos</atitle><jtitle>Biology of reproduction</jtitle><addtitle>Biol Reprod</addtitle><date>2002-02-01</date><risdate>2002</risdate><volume>66</volume><issue>2</issue><spage>534</spage><epage>543</epage><pages>534-543</pages><issn>0006-3363</issn><eissn>1529-7268</eissn><coden>BIREBV</coden><abstract>This study focused on nucleolar changes in bovine embryos reconstructed from enucleated mature oocytes fused with blastomeres
of morulae or with cultured, serum unstarved bovine fetal skin fibroblasts (embryonic vs. somatic cloning). The nucleotransferred
(NT) embryos were collected and fixed at time intervals of 1â2 h (early 1-cell stage), 10â15 h (late 1-cell stage), 22â24
h (2-cell stage), 37â38 h (4-cell stage), 40â41 h (early 8-cell stage), 47â48 h (late 8-cell stage), and 55 h (16-cell stage)
after fusion. Immunocytochemistry by light and electron microscopy was used for structure-function characterization of nucleolar
components. Antibodies against RNA, protein B23, protein C23, and fibrillarin were applied. In addition, DNA was localized
by the terminal deoxynucleotidyl transferase (TdT) technique, and the functional organization of chromatin was determined
with the nick-translation immunogold approach. The results show that fully reticulated (active) nucleoli observed in donor
cells immediately before fusion as well as in the early 1-cell stage after fusion were progressively transformed into nucleolar
bodies displaying decreasing numbers of vacuoles from the 2- to 4-cell stage in both types of reconstructed embryos. At the
late 8-cell stage, morphological signs of resuming nucleolar activity were detected. Numerous new small vacuoles appeared,
and chromatin blocks reassociated with the nucleolar body. During this period, nick-translation technique revealed numerous
active DNA sites in the periphery of chromatin blocks associated with the nucleolar body. Fully reticulated nucleoli were
again observed as early as the 16-cell stage of embryonic cloned embryos. In comparison, the embryos obtained by fetal cloning
displayed a lower tendency to develop, mainly during the first cell cycle and during the period of presumed reactivation.
Correlatively, the changes in nucleolar morphology (desegregation and rebuilding) were at least delayed in many somatic NT
embryos in comparison with the embryonic NT group. It is concluded that complete reprogramming of rRNA gene expression is
part of the general nuclear reprogramming necessary for development after NT.</abstract><cop>Madison, WI</cop><pub>Society for the Study of Reproduction</pub><pmid>11804972</pmid><doi>10.1095/biolreprod66.2.534</doi><tpages>10</tpages></addata></record> |
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| ispartof | Biology of reproduction, 2002-02, Vol.66 (2), p.534-543 |
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| language | eng |
| recordid | cdi_hal_primary_oai_HAL_hal_02671371v1 |
| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; BioOne Complete; Oxford University Press Journals All Titles (1996-Current) |
| subjects | Animals Antibiotics, Antineoplastic - pharmacology Antibodies, Monoclonal - immunology Biological and medical sciences Cattle Cell Fusion Cell Nucleolus - genetics Cell Nucleolus - metabolism Cell Nucleolus - ultrastructure Cloning, Organism Dactinomycin - pharmacology DNA - metabolism Embryo Transfer Embryonic and Fetal Development - physiology Female Fibroblasts - physiology Fluorescent Antibody Technique Gene Expression Regulation, Developmental - genetics Gene Expression Regulation, Developmental - physiology In Situ Nick-End Labeling Life Sciences Microscopy, Confocal Microscopy, Electron Microscopy, Fluorescence Oocytes - physiology Pregnancy |
| title | Nucleolar Changes in Bovine Nucleotransferred Embryos |
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