Enzymes involved in the biosynthesis of the very-long-chain polyunsaturated fatty acids of rat spermatogenic cell sphingolipids

Autores
Santiago Valtierra, Florencia Ximena; Oresti, Gerardo Martin; Aveldaño, Marta Isabel
Año de publicación
2017
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
The sphingolipids (SL) of rodent germ cells contain very-long- chain polyunsaturated fatty acids (V), in nonhydroxy (n-V) and 2-hy- droxy (h-V) forms, whose biosynthesis requires the expression of several elongases (Elovl) and a fatty acid 2-hydroxylase (Fa2h). Our objective was to characterize the expression of these enzymes as a function of postnatal development and germ cell differentiation. We employed qPCR for mRNA levels, Western blot and immunofluo- rescence for protein expression, and [3 H]-labeled precursors for en- zyme activity. Elovl4, involved in C24-C34 fatty acid biosynthesis, was expressed at the mRNA, but not at the protein level, at early prepu- beral ages. Such mRNA was a product of Sertoli cells. Elovl4 mRNA and protein were both produced by germ cells. In agreement with the relative abundance of n-V in their SL, the Elovl4 enzymatic ac- tivity was higher in the premeiotic pachytene spermatocytes than in postmeiotic round and late spermatids (p<0.05), and was negligible in Sertoli cells. By contrast to Elovl4 mRNA, that of Fa2h was absent from Sertoli cells and from prepuberal testes (p<0.05). Fa2h mRNA and protein were detectable only in concomitance with the appear- ance of spermatids, the sperm precursors whose SL are the richest in h-V. Consistently, selective depletion of germ (but not Sertoli) cells from adult rat testes by exposures to mild hyperthermia reduced the mRNA levels of Fa2h to a much larger extent (p<0.001) than that of Elovl4. Among germ cells of adult testes, Elovl4 protein content was high in spermatocytes and late spermatids, while Fa2h was mostly expressed in the latter, residual bodies and spermatozoa. By using inhibitors of specific steps of the SL biosynthetic route, germ cells in culture showed ability to de novo synthesize SL containing n-V and h-V. Our results underscore the presence of a developmentally programmed and a cell-specific regulation of the Elovl4 and Fa2h expression and activity as germ cell differentiation proceeds.
Fil: Santiago Valtierra, Florencia Ximena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Oresti, Gerardo Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Aveldaño, Marta Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
LXII Reunión Científica de la Sociedad Argentina de Investigación Clínica; LIII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica ; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología
Buenos Aires
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Investigación Bioquímica y Biología Molecular
Sociedad Argentina de Inmunología
Sociedad Argentina de Andrología
Sociedad Argentina de Biofísica
Sociedad Argentina de Biología
Sociedad Argentina de Farmacología Experimental
Sociedad Argentina de Fisiologia
Sociedad Argentina de Hematología
Sociedad Argentina de Protozoología
Materia
spermatogenic cells
sphingolipids
very-long-chain
PUFA
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/233184
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/233184
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Enzymes involved in the biosynthesis of the very-long-chain polyunsaturated fatty acids of rat spermatogenic cell sphingolipidsSantiago Valtierra, Florencia XimenaOresti, Gerardo MartinAveldaño, Marta Isabelspermatogenic cellssphingolipidsvery-long-chainPUFAhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The sphingolipids (SL) of rodent germ cells contain very-long- chain polyunsaturated fatty acids (V), in nonhydroxy (n-V) and 2-hy- droxy (h-V) forms, whose biosynthesis requires the expression of several elongases (Elovl) and a fatty acid 2-hydroxylase (Fa2h). Our objective was to characterize the expression of these enzymes as a function of postnatal development and germ cell differentiation. We employed qPCR for mRNA levels, Western blot and immunofluo- rescence for protein expression, and [3 H]-labeled precursors for en- zyme activity. Elovl4, involved in C24-C34 fatty acid biosynthesis, was expressed at the mRNA, but not at the protein level, at early prepu- beral ages. Such mRNA was a product of Sertoli cells. Elovl4 mRNA and protein were both produced by germ cells. In agreement with the relative abundance of n-V in their SL, the Elovl4 enzymatic ac- tivity was higher in the premeiotic pachytene spermatocytes than in postmeiotic round and late spermatids (p<0.05), and was negligible in Sertoli cells. By contrast to Elovl4 mRNA, that of Fa2h was absent from Sertoli cells and from prepuberal testes (p<0.05). Fa2h mRNA and protein were detectable only in concomitance with the appear- ance of spermatids, the sperm precursors whose SL are the richest in h-V. Consistently, selective depletion of germ (but not Sertoli) cells from adult rat testes by exposures to mild hyperthermia reduced the mRNA levels of Fa2h to a much larger extent (p<0.001) than that of Elovl4. Among germ cells of adult testes, Elovl4 protein content was high in spermatocytes and late spermatids, while Fa2h was mostly expressed in the latter, residual bodies and spermatozoa. By using inhibitors of specific steps of the SL biosynthetic route, germ cells in culture showed ability to de novo synthesize SL containing n-V and h-V. Our results underscore the presence of a developmentally programmed and a cell-specific regulation of the Elovl4 and Fa2h expression and activity as germ cell differentiation proceeds.Fil: Santiago Valtierra, Florencia Ximena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaFil: Oresti, Gerardo Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaFil: Aveldaño, Marta Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaLXII Reunión Científica de la Sociedad Argentina de Investigación Clínica; LIII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica ; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de ProtozoologíaBuenos AiresArgentinaSociedad Argentina de Investigación ClínicaSociedad Argentina de Investigación Bioquímica y Biología MolecularSociedad Argentina de InmunologíaSociedad Argentina de AndrologíaSociedad Argentina de BiofísicaSociedad Argentina de BiologíaSociedad Argentina de Farmacología ExperimentalSociedad Argentina de FisiologiaSociedad Argentina de HematologíaSociedad Argentina de ProtozoologíaFundación Revista medicina2017info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/233184Enzymes involved in the biosynthesis of the very-long-chain polyunsaturated fatty acids of rat spermatogenic cell sphingolipids; LXII Reunión Científica de la Sociedad Argentina de Investigación Clínica; LIII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica ; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología; Buenos Aires; Argentina; 2017; 122-1230025-76801669-9106CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.saic.org.ar/revista-medicinaNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-10T13:05:34Zoai:ri.conicet.gov.ar:11336/233184instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-10 13:05:34.511CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Enzymes involved in the biosynthesis of the very-long-chain polyunsaturated fatty acids of rat spermatogenic cell sphingolipids
title Enzymes involved in the biosynthesis of the very-long-chain polyunsaturated fatty acids of rat spermatogenic cell sphingolipids
spellingShingle Enzymes involved in the biosynthesis of the very-long-chain polyunsaturated fatty acids of rat spermatogenic cell sphingolipids
Santiago Valtierra, Florencia Ximena
spermatogenic cells
sphingolipids
very-long-chain
PUFA
title_short Enzymes involved in the biosynthesis of the very-long-chain polyunsaturated fatty acids of rat spermatogenic cell sphingolipids
title_full Enzymes involved in the biosynthesis of the very-long-chain polyunsaturated fatty acids of rat spermatogenic cell sphingolipids
title_fullStr Enzymes involved in the biosynthesis of the very-long-chain polyunsaturated fatty acids of rat spermatogenic cell sphingolipids
title_full_unstemmed Enzymes involved in the biosynthesis of the very-long-chain polyunsaturated fatty acids of rat spermatogenic cell sphingolipids
title_sort Enzymes involved in the biosynthesis of the very-long-chain polyunsaturated fatty acids of rat spermatogenic cell sphingolipids
dc.creator.none.fl_str_mv Santiago Valtierra, Florencia Ximena
Oresti, Gerardo Martin
Aveldaño, Marta Isabel
author Santiago Valtierra, Florencia Ximena
author_facet Santiago Valtierra, Florencia Ximena
Oresti, Gerardo Martin
Aveldaño, Marta Isabel
author_role author
author2 Oresti, Gerardo Martin
Aveldaño, Marta Isabel
author2_role author
author
dc.subject.none.fl_str_mv spermatogenic cells
sphingolipids
very-long-chain
PUFA
topic spermatogenic cells
sphingolipids
very-long-chain
PUFA
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The sphingolipids (SL) of rodent germ cells contain very-long- chain polyunsaturated fatty acids (V), in nonhydroxy (n-V) and 2-hy- droxy (h-V) forms, whose biosynthesis requires the expression of several elongases (Elovl) and a fatty acid 2-hydroxylase (Fa2h). Our objective was to characterize the expression of these enzymes as a function of postnatal development and germ cell differentiation. We employed qPCR for mRNA levels, Western blot and immunofluo- rescence for protein expression, and [3 H]-labeled precursors for en- zyme activity. Elovl4, involved in C24-C34 fatty acid biosynthesis, was expressed at the mRNA, but not at the protein level, at early prepu- beral ages. Such mRNA was a product of Sertoli cells. Elovl4 mRNA and protein were both produced by germ cells. In agreement with the relative abundance of n-V in their SL, the Elovl4 enzymatic ac- tivity was higher in the premeiotic pachytene spermatocytes than in postmeiotic round and late spermatids (p<0.05), and was negligible in Sertoli cells. By contrast to Elovl4 mRNA, that of Fa2h was absent from Sertoli cells and from prepuberal testes (p<0.05). Fa2h mRNA and protein were detectable only in concomitance with the appear- ance of spermatids, the sperm precursors whose SL are the richest in h-V. Consistently, selective depletion of germ (but not Sertoli) cells from adult rat testes by exposures to mild hyperthermia reduced the mRNA levels of Fa2h to a much larger extent (p<0.001) than that of Elovl4. Among germ cells of adult testes, Elovl4 protein content was high in spermatocytes and late spermatids, while Fa2h was mostly expressed in the latter, residual bodies and spermatozoa. By using inhibitors of specific steps of the SL biosynthetic route, germ cells in culture showed ability to de novo synthesize SL containing n-V and h-V. Our results underscore the presence of a developmentally programmed and a cell-specific regulation of the Elovl4 and Fa2h expression and activity as germ cell differentiation proceeds.
Fil: Santiago Valtierra, Florencia Ximena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Oresti, Gerardo Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Aveldaño, Marta Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
LXII Reunión Científica de la Sociedad Argentina de Investigación Clínica; LIII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica ; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología
Buenos Aires
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Investigación Bioquímica y Biología Molecular
Sociedad Argentina de Inmunología
Sociedad Argentina de Andrología
Sociedad Argentina de Biofísica
Sociedad Argentina de Biología
Sociedad Argentina de Farmacología Experimental
Sociedad Argentina de Fisiologia
Sociedad Argentina de Hematología
Sociedad Argentina de Protozoología
description The sphingolipids (SL) of rodent germ cells contain very-long- chain polyunsaturated fatty acids (V), in nonhydroxy (n-V) and 2-hy- droxy (h-V) forms, whose biosynthesis requires the expression of several elongases (Elovl) and a fatty acid 2-hydroxylase (Fa2h). Our objective was to characterize the expression of these enzymes as a function of postnatal development and germ cell differentiation. We employed qPCR for mRNA levels, Western blot and immunofluo- rescence for protein expression, and [3 H]-labeled precursors for en- zyme activity. Elovl4, involved in C24-C34 fatty acid biosynthesis, was expressed at the mRNA, but not at the protein level, at early prepu- beral ages. Such mRNA was a product of Sertoli cells. Elovl4 mRNA and protein were both produced by germ cells. In agreement with the relative abundance of n-V in their SL, the Elovl4 enzymatic ac- tivity was higher in the premeiotic pachytene spermatocytes than in postmeiotic round and late spermatids (p<0.05), and was negligible in Sertoli cells. By contrast to Elovl4 mRNA, that of Fa2h was absent from Sertoli cells and from prepuberal testes (p<0.05). Fa2h mRNA and protein were detectable only in concomitance with the appear- ance of spermatids, the sperm precursors whose SL are the richest in h-V. Consistently, selective depletion of germ (but not Sertoli) cells from adult rat testes by exposures to mild hyperthermia reduced the mRNA levels of Fa2h to a much larger extent (p<0.001) than that of Elovl4. Among germ cells of adult testes, Elovl4 protein content was high in spermatocytes and late spermatids, while Fa2h was mostly expressed in the latter, residual bodies and spermatozoa. By using inhibitors of specific steps of the SL biosynthetic route, germ cells in culture showed ability to de novo synthesize SL containing n-V and h-V. Our results underscore the presence of a developmentally programmed and a cell-specific regulation of the Elovl4 and Fa2h expression and activity as germ cell differentiation proceeds.
publishDate 2017
dc.date.none.fl_str_mv 2017
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Congreso
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/233184
Enzymes involved in the biosynthesis of the very-long-chain polyunsaturated fatty acids of rat spermatogenic cell sphingolipids; LXII Reunión Científica de la Sociedad Argentina de Investigación Clínica; LIII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica ; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología; Buenos Aires; Argentina; 2017; 122-123
0025-7680
1669-9106
CONICET Digital
CONICET
url http://hdl.handle.net/11336/233184
identifier_str_mv Enzymes involved in the biosynthesis of the very-long-chain polyunsaturated fatty acids of rat spermatogenic cell sphingolipids; LXII Reunión Científica de la Sociedad Argentina de Investigación Clínica; LIII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica ; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología; Buenos Aires; Argentina; 2017; 122-123
0025-7680
1669-9106
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.saic.org.ar/revista-medicina
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Fundación Revista medicina
publisher.none.fl_str_mv Fundación Revista medicina
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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