Specific deletion of AMP-activated protein kinase (α1AMPK) in mouse Sertoli cells modifies germ cell quality.
Identifieur interne : 002F98 ( Ncbi/Merge ); précédent : 002F97; suivant : 002F99Specific deletion of AMP-activated protein kinase (α1AMPK) in mouse Sertoli cells modifies germ cell quality.
Auteurs : Michael J. Bertoldo [Australie] ; Edith Guibert [France] ; Melanie Faure [France] ; Florian Guillou [France] ; Christelle Ramé [France] ; Lydie Nadal-Desbarats [France] ; Marc Foretz [France] ; Benoit Viollet [France] ; Joëlle Dupont [France] ; Pascal Froment [France]Source :
- Molecular and cellular endocrinology [ 1872-8057 ] ; 2016.
Descripteurs français
- KwdFr :
- AMP-Activated Protein Kinases (génétique), AMP-Activated Protein Kinases (métabolisme), Acrosome (enzymologie), Adénosine triphosphate (biosynthèse), Animaux, Cellules cultivées, Cellules de Sertoli (enzymologie), Expression des gènes, Femelle, Infertilité masculine (enzymologie), Infertilité masculine (génétique), Motilité des spermatozoïdes, Mâle, Prolifération cellulaire, Sirtuine-1 (génétique), Sirtuine-1 (métabolisme), Souris de lignée C57BL, Souris knockout, Survie cellulaire, Transport de protéines.
- MESH :
- biosynthèse : Adénosine triphosphate.
- enzymologie : Acrosome, Cellules de Sertoli, Infertilité masculine.
- génétique : AMP-Activated Protein Kinases, Infertilité masculine, Sirtuine-1.
- métabolisme : AMP-Activated Protein Kinases, Sirtuine-1.
- Animaux, Cellules cultivées, Expression des gènes, Femelle, Motilité des spermatozoïdes, Mâle, Prolifération cellulaire, Souris de lignée C57BL, Souris knockout, Survie cellulaire, Transport de protéines.
English descriptors
- KwdEn :
- AMP-Activated Protein Kinases (genetics), AMP-Activated Protein Kinases (metabolism), Acrosome (enzymology), Adenosine Triphosphate (biosynthesis), Animals, Cell Proliferation, Cell Survival, Cells, Cultured, Female, Gene Expression, Infertility, Male (enzymology), Infertility, Male (genetics), Male, Mice, Inbred C57BL, Mice, Knockout, Protein Transport, Sertoli Cells (enzymology), Sirtuin 1 (genetics), Sirtuin 1 (metabolism), Sperm Motility.
- MESH :
- chemical , biosynthesis : Adenosine Triphosphate.
- chemical , genetics : AMP-Activated Protein Kinases, Sirtuin 1.
- chemical , metabolism : AMP-Activated Protein Kinases, Sirtuin 1.
- enzymology : Acrosome, Infertility, Male, Sertoli Cells.
- genetics : Infertility, Male.
- Animals, Cell Proliferation, Cell Survival, Cells, Cultured, Female, Gene Expression, Male, Mice, Inbred C57BL, Mice, Knockout, Protein Transport, Sperm Motility.
Abstract
The AMP-activated protein kinase (AMPK) is an important regulator of cellular energy homeostasis which plays a role in fertility. Complete disruption of the AMPK catalytic subunit α1 gene (α1AMPK KO) in male mice results in a decrease in litter size which is associated with the production of altered sperm morphology and motility. Because of the importance of Sertoli cells in the formation of germ cells, we have chosen to selectively disrupt α1AMPK only in the Sertoli cells in mice (Sc-α1AMPK-KO mice). Specific deletion of the α1AMPK gene in Sertoli cells resulted in a 25% reduction in male fertility associated with abnormal spermatozoa with a thin head. No clear alterations in testis morphology or modification in the number of Sertoli cells in vivo were observed, but a dysregulation in energy metabolism in Sertoli cells occurred. We have reported an increase in lactate production, in lipid droplets, and a reduction in ATP production in Sc-α1AMPK-KO Sertoli cells. These perturbations were associated with lower expression of mitochondrial markers (cytochrome c and PGC1-α). In addition another metabolic sensor, the deacetylase SIRT1, had a reduction in expression which is correlated with a decline in deacetylase activity. Finally, expression and localization of junctions forming the blood-testis barrier between Sertoli cells themselves and with germ cells were deregulated in Sc-α1AMPK-KO. In conclusion, these results suggest that dysregulation of the energy sensing machinery exclusively through disruption of α1AMPK in Sertoli cells translates to a reduction in the quality of germ cells and fertility.
DOI: 10.1016/j.mce.2016.01.001
PubMed: 26772142
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<author><name sortKey="Nadal Desbarats, Lydie" sort="Nadal Desbarats, Lydie" uniqKey="Nadal Desbarats L" first="Lydie" last="Nadal-Desbarats">Lydie Nadal-Desbarats</name>
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<term>AMP-Activated Protein Kinases (metabolism)</term>
<term>Acrosome (enzymology)</term>
<term>Adenosine Triphosphate (biosynthesis)</term>
<term>Animals</term>
<term>Cell Proliferation</term>
<term>Cell Survival</term>
<term>Cells, Cultured</term>
<term>Female</term>
<term>Gene Expression</term>
<term>Infertility, Male (enzymology)</term>
<term>Infertility, Male (genetics)</term>
<term>Male</term>
<term>Mice, Inbred C57BL</term>
<term>Mice, Knockout</term>
<term>Protein Transport</term>
<term>Sertoli Cells (enzymology)</term>
<term>Sirtuin 1 (genetics)</term>
<term>Sirtuin 1 (metabolism)</term>
<term>Sperm Motility</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>AMP-Activated Protein Kinases (génétique)</term>
<term>AMP-Activated Protein Kinases (métabolisme)</term>
<term>Acrosome (enzymologie)</term>
<term>Adénosine triphosphate (biosynthèse)</term>
<term>Animaux</term>
<term>Cellules cultivées</term>
<term>Cellules de Sertoli (enzymologie)</term>
<term>Expression des gènes</term>
<term>Femelle</term>
<term>Infertilité masculine (enzymologie)</term>
<term>Infertilité masculine (génétique)</term>
<term>Motilité des spermatozoïdes</term>
<term>Mâle</term>
<term>Prolifération cellulaire</term>
<term>Sirtuine-1 (génétique)</term>
<term>Sirtuine-1 (métabolisme)</term>
<term>Souris de lignée C57BL</term>
<term>Souris knockout</term>
<term>Survie cellulaire</term>
<term>Transport de protéines</term>
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<term>Sirtuin 1</term>
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<term>Sirtuin 1</term>
</keywords>
<keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>Adénosine triphosphate</term>
</keywords>
<keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Acrosome</term>
<term>Cellules de Sertoli</term>
<term>Infertilité masculine</term>
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<keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Acrosome</term>
<term>Infertility, Male</term>
<term>Sertoli Cells</term>
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</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>AMP-Activated Protein Kinases</term>
<term>Infertilité masculine</term>
<term>Sirtuine-1</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>AMP-Activated Protein Kinases</term>
<term>Sirtuine-1</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>Cell Proliferation</term>
<term>Cell Survival</term>
<term>Cells, Cultured</term>
<term>Female</term>
<term>Gene Expression</term>
<term>Male</term>
<term>Mice, Inbred C57BL</term>
<term>Mice, Knockout</term>
<term>Protein Transport</term>
<term>Sperm Motility</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Animaux</term>
<term>Cellules cultivées</term>
<term>Expression des gènes</term>
<term>Femelle</term>
<term>Motilité des spermatozoïdes</term>
<term>Mâle</term>
<term>Prolifération cellulaire</term>
<term>Souris de lignée C57BL</term>
<term>Souris knockout</term>
<term>Survie cellulaire</term>
<term>Transport de protéines</term>
</keywords>
</textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en">The AMP-activated protein kinase (AMPK) is an important regulator of cellular energy homeostasis which plays a role in fertility. Complete disruption of the AMPK catalytic subunit α1 gene (α1AMPK KO) in male mice results in a decrease in litter size which is associated with the production of altered sperm morphology and motility. Because of the importance of Sertoli cells in the formation of germ cells, we have chosen to selectively disrupt α1AMPK only in the Sertoli cells in mice (Sc-α1AMPK-KO mice). Specific deletion of the α1AMPK gene in Sertoli cells resulted in a 25% reduction in male fertility associated with abnormal spermatozoa with a thin head. No clear alterations in testis morphology or modification in the number of Sertoli cells in vivo were observed, but a dysregulation in energy metabolism in Sertoli cells occurred. We have reported an increase in lactate production, in lipid droplets, and a reduction in ATP production in Sc-α1AMPK-KO Sertoli cells. These perturbations were associated with lower expression of mitochondrial markers (cytochrome c and PGC1-α). In addition another metabolic sensor, the deacetylase SIRT1, had a reduction in expression which is correlated with a decline in deacetylase activity. Finally, expression and localization of junctions forming the blood-testis barrier between Sertoli cells themselves and with germ cells were deregulated in Sc-α1AMPK-KO. In conclusion, these results suggest that dysregulation of the energy sensing machinery exclusively through disruption of α1AMPK in Sertoli cells translates to a reduction in the quality of germ cells and fertility.</div>
</front>
</TEI>
<pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">26772142</PMID>
<DateCreated><Year>2016</Year>
<Month>02</Month>
<Day>12</Day>
</DateCreated>
<DateCompleted><Year>2016</Year>
<Month>11</Month>
<Day>04</Day>
</DateCompleted>
<DateRevised><Year>2017</Year>
<Month>11</Month>
<Day>16</Day>
</DateRevised>
<Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1872-8057</ISSN>
<JournalIssue CitedMedium="Internet"><Volume>423</Volume>
<PubDate><Year>2016</Year>
<Month>Mar</Month>
<Day>05</Day>
</PubDate>
</JournalIssue>
<Title>Molecular and cellular endocrinology</Title>
<ISOAbbreviation>Mol. Cell. Endocrinol.</ISOAbbreviation>
</Journal>
<ArticleTitle>Specific deletion of AMP-activated protein kinase (α1AMPK) in mouse Sertoli cells modifies germ cell quality.</ArticleTitle>
<Pagination><MedlinePgn>96-112</MedlinePgn>
</Pagination>
<ELocationID EIdType="doi" ValidYN="Y">10.1016/j.mce.2016.01.001</ELocationID>
<ELocationID EIdType="pii" ValidYN="Y">S0303-7207(16)30001-6</ELocationID>
<Abstract><AbstractText>The AMP-activated protein kinase (AMPK) is an important regulator of cellular energy homeostasis which plays a role in fertility. Complete disruption of the AMPK catalytic subunit α1 gene (α1AMPK KO) in male mice results in a decrease in litter size which is associated with the production of altered sperm morphology and motility. Because of the importance of Sertoli cells in the formation of germ cells, we have chosen to selectively disrupt α1AMPK only in the Sertoli cells in mice (Sc-α1AMPK-KO mice). Specific deletion of the α1AMPK gene in Sertoli cells resulted in a 25% reduction in male fertility associated with abnormal spermatozoa with a thin head. No clear alterations in testis morphology or modification in the number of Sertoli cells in vivo were observed, but a dysregulation in energy metabolism in Sertoli cells occurred. We have reported an increase in lactate production, in lipid droplets, and a reduction in ATP production in Sc-α1AMPK-KO Sertoli cells. These perturbations were associated with lower expression of mitochondrial markers (cytochrome c and PGC1-α). In addition another metabolic sensor, the deacetylase SIRT1, had a reduction in expression which is correlated with a decline in deacetylase activity. Finally, expression and localization of junctions forming the blood-testis barrier between Sertoli cells themselves and with germ cells were deregulated in Sc-α1AMPK-KO. In conclusion, these results suggest that dysregulation of the energy sensing machinery exclusively through disruption of α1AMPK in Sertoli cells translates to a reduction in the quality of germ cells and fertility.</AbstractText>
<CopyrightInformation>Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.</CopyrightInformation>
</Abstract>
<AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Bertoldo</LastName>
<ForeName>Michael J</ForeName>
<Initials>MJ</Initials>
<AffiliationInfo><Affiliation>Unité de Physiologie de la Reproduction et des Comportements, Institut National de la Recherche Agronomique, Centre Val de Loire, UMR85, 37380 Nouzilly, France; School of Women's and Children's Health, Discipline of Obstetrics and Gynaecology, University of New South Wales, Sydney, NSW, Australia.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Guibert</LastName>
<ForeName>Edith</ForeName>
<Initials>E</Initials>
<AffiliationInfo><Affiliation>Unité de Physiologie de la Reproduction et des Comportements, Institut National de la Recherche Agronomique, Centre Val de Loire, UMR85, 37380 Nouzilly, France.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Faure</LastName>
<ForeName>Melanie</ForeName>
<Initials>M</Initials>
<AffiliationInfo><Affiliation>Unité de Physiologie de la Reproduction et des Comportements, Institut National de la Recherche Agronomique, Centre Val de Loire, UMR85, 37380 Nouzilly, France.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Guillou</LastName>
<ForeName>Florian</ForeName>
<Initials>F</Initials>
<AffiliationInfo><Affiliation>Unité de Physiologie de la Reproduction et des Comportements, Institut National de la Recherche Agronomique, Centre Val de Loire, UMR85, 37380 Nouzilly, France.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Ramé</LastName>
<ForeName>Christelle</ForeName>
<Initials>C</Initials>
<AffiliationInfo><Affiliation>Unité de Physiologie de la Reproduction et des Comportements, Institut National de la Recherche Agronomique, Centre Val de Loire, UMR85, 37380 Nouzilly, France.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Nadal-Desbarats</LastName>
<ForeName>Lydie</ForeName>
<Initials>L</Initials>
<AffiliationInfo><Affiliation>INSERM U930, Équipe Neurogénétique et Neurométabolomique, Université François-Rabelais, 37044 Tours, France; Département d'Analyse Chimique Biologique et Médicale, PPF "Analyses des Systèmes Biologiques", Université François-Rabelais, Tours, France.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Foretz</LastName>
<ForeName>Marc</ForeName>
<Initials>M</Initials>
<AffiliationInfo><Affiliation>INSERM, U1016, Institut Cochin, Paris, France; CNRS, UMR8104, Paris, France; Univ Paris Descartes, Sorbonne Paris Cité, Paris, France.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Viollet</LastName>
<ForeName>Benoit</ForeName>
<Initials>B</Initials>
<AffiliationInfo><Affiliation>INSERM, U1016, Institut Cochin, Paris, France; CNRS, UMR8104, Paris, France; Univ Paris Descartes, Sorbonne Paris Cité, Paris, France.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Dupont</LastName>
<ForeName>Joëlle</ForeName>
<Initials>J</Initials>
<AffiliationInfo><Affiliation>Unité de Physiologie de la Reproduction et des Comportements, Institut National de la Recherche Agronomique, Centre Val de Loire, UMR85, 37380 Nouzilly, France.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Froment</LastName>
<ForeName>Pascal</ForeName>
<Initials>P</Initials>
<AffiliationInfo><Affiliation>Unité de Physiologie de la Reproduction et des Comportements, Institut National de la Recherche Agronomique, Centre Val de Loire, UMR85, 37380 Nouzilly, France. Electronic address: pascal.froment@tours.inra.fr.</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>2016</Year>
<Month>01</Month>
<Day>07</Day>
</ArticleDate>
</Article>
<MedlineJournalInfo><Country>Ireland</Country>
<MedlineTA>Mol Cell Endocrinol</MedlineTA>
<NlmUniqueID>7500844</NlmUniqueID>
<ISSNLinking>0303-7207</ISSNLinking>
</MedlineJournalInfo>
<ChemicalList><Chemical><RegistryNumber>8L70Q75FXE</RegistryNumber>
<NameOfSubstance UI="D000255">Adenosine Triphosphate</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>EC 2.7.11.1</RegistryNumber>
<NameOfSubstance UI="D055372">AMP-Activated Protein Kinases</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>EC 3.5.1.-</RegistryNumber>
<NameOfSubstance UI="C470545">Sirt1 protein, mouse</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>EC 3.5.1.-</RegistryNumber>
<NameOfSubstance UI="D056564">Sirtuin 1</NameOfSubstance>
</Chemical>
</ChemicalList>
<CitationSubset>IM</CitationSubset>
<MeshHeadingList><MeshHeading><DescriptorName UI="D055372" MajorTopicYN="N">AMP-Activated Protein Kinases</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName>
<QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D000177" MajorTopicYN="N">Acrosome</DescriptorName>
<QualifierName UI="Q000201" MajorTopicYN="Y">enzymology</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D000255" MajorTopicYN="N">Adenosine Triphosphate</DescriptorName>
<QualifierName UI="Q000096" MajorTopicYN="N">biosynthesis</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D000818" MajorTopicYN="N">Animals</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D049109" MajorTopicYN="N">Cell Proliferation</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D002470" MajorTopicYN="N">Cell Survival</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D002478" MajorTopicYN="N">Cells, Cultured</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D005260" MajorTopicYN="N">Female</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D015870" MajorTopicYN="N">Gene Expression</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D007248" MajorTopicYN="N">Infertility, Male</DescriptorName>
<QualifierName UI="Q000201" MajorTopicYN="N">enzymology</QualifierName>
<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D008297" MajorTopicYN="N">Male</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D008810" MajorTopicYN="N">Mice, Inbred C57BL</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D018345" MajorTopicYN="N">Mice, Knockout</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D021381" MajorTopicYN="N">Protein Transport</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D012708" MajorTopicYN="N">Sertoli Cells</DescriptorName>
<QualifierName UI="Q000201" MajorTopicYN="Y">enzymology</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D056564" MajorTopicYN="N">Sirtuin 1</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
<QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D013081" MajorTopicYN="N">Sperm Motility</DescriptorName>
</MeshHeading>
</MeshHeadingList>
<KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">AMP-activated protein kinase</Keyword>
<Keyword MajorTopicYN="N">Blood testis barrier</Keyword>
<Keyword MajorTopicYN="N">Fertility</Keyword>
<Keyword MajorTopicYN="N">Metabolism</Keyword>
<Keyword MajorTopicYN="N">Sertoli cells</Keyword>
<Keyword MajorTopicYN="N">Testis</Keyword>
</KeywordList>
</MedlineCitation>
<PubmedData><History><PubMedPubDate PubStatus="received"><Year>2015</Year>
<Month>10</Month>
<Day>01</Day>
</PubMedPubDate>
<PubMedPubDate PubStatus="revised"><Year>2015</Year>
<Month>12</Month>
<Day>15</Day>
</PubMedPubDate>
<PubMedPubDate PubStatus="accepted"><Year>2016</Year>
<Month>01</Month>
<Day>04</Day>
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<Month>1</Month>
<Day>17</Day>
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<ArticleId IdType="doi">10.1016/j.mce.2016.01.001</ArticleId>
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<affiliations><list><country><li>Australie</li>
<li>France</li>
</country>
<region><li>Centre-Val de Loire</li>
<li>Région Centre</li>
<li>Île-de-France</li>
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<settlement><li>Paris</li>
<li>Tours</li>
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<tree><country name="Australie"><noRegion><name sortKey="Bertoldo, Michael J" sort="Bertoldo, Michael J" uniqKey="Bertoldo M" first="Michael J" last="Bertoldo">Michael J. Bertoldo</name>
</noRegion>
</country>
<country name="France"><noRegion><name sortKey="Guibert, Edith" sort="Guibert, Edith" uniqKey="Guibert E" first="Edith" last="Guibert">Edith Guibert</name>
</noRegion>
<name sortKey="Dupont, Joelle" sort="Dupont, Joelle" uniqKey="Dupont J" first="Joëlle" last="Dupont">Joëlle Dupont</name>
<name sortKey="Faure, Melanie" sort="Faure, Melanie" uniqKey="Faure M" first="Melanie" last="Faure">Melanie Faure</name>
<name sortKey="Foretz, Marc" sort="Foretz, Marc" uniqKey="Foretz M" first="Marc" last="Foretz">Marc Foretz</name>
<name sortKey="Froment, Pascal" sort="Froment, Pascal" uniqKey="Froment P" first="Pascal" last="Froment">Pascal Froment</name>
<name sortKey="Guillou, Florian" sort="Guillou, Florian" uniqKey="Guillou F" first="Florian" last="Guillou">Florian Guillou</name>
<name sortKey="Nadal Desbarats, Lydie" sort="Nadal Desbarats, Lydie" uniqKey="Nadal Desbarats L" first="Lydie" last="Nadal-Desbarats">Lydie Nadal-Desbarats</name>
<name sortKey="Rame, Christelle" sort="Rame, Christelle" uniqKey="Rame C" first="Christelle" last="Ramé">Christelle Ramé</name>
<name sortKey="Viollet, Benoit" sort="Viollet, Benoit" uniqKey="Viollet B" first="Benoit" last="Viollet">Benoit Viollet</name>
</country>
</tree>
</affiliations>
</record>
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