Transcriptome profile in the skeletal muscle of cattle progeny as a function of maternal protein supplementation during mid-gestation

This study aimed to evaluate the differentially expressed genes (DEG), their biological functions, and the main predictor genes in skeletal muscle of the offspring resulting from dams supplemented or not with protein during mid-gestation. From 100 to 200 days of gestation, 10 Tabapuã cows pregnant o...

Full description

Autores:
Carvalho, Elisa
Costa, Thaís C.
Sanglard, Leticia P.
Nascimento, Karolina B.
Moreno Meneses, Javier Andrés
Galvão, Matheus C.
Serão, Nick V.L.
Duarte, Marcio S.
Pies Gionbelli, Mateus
Tipo de recurso:
Article of investigation
Fecha de publicación:
2022
Institución:
Universidad de Ciencias Aplicadas y Ambientales U.D.C.A
Repositorio:
Repositorio Institucional UDCA
Idioma:
eng
OAI Identifier:
oai:repository.udca.edu.co:11158/4821
Acceso en línea:
https://repository.udca.edu.co/handle/11158/4821
Palabra clave:
Ganado bovino
Transcriptomas
Progenie
Gestación
Rights
openAccess
License
https://creativecommons.org/licenses/by-nc-sa/4.0/legalcode.es
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repository_id_str
dc.title.eng.fl_str_mv Transcriptome profile in the skeletal muscle of cattle progeny as a function of maternal protein supplementation during mid-gestation
title Transcriptome profile in the skeletal muscle of cattle progeny as a function of maternal protein supplementation during mid-gestation
spellingShingle Transcriptome profile in the skeletal muscle of cattle progeny as a function of maternal protein supplementation during mid-gestation
Ganado bovino
Transcriptomas
Progenie
Gestación
title_short Transcriptome profile in the skeletal muscle of cattle progeny as a function of maternal protein supplementation during mid-gestation
title_full Transcriptome profile in the skeletal muscle of cattle progeny as a function of maternal protein supplementation during mid-gestation
title_fullStr Transcriptome profile in the skeletal muscle of cattle progeny as a function of maternal protein supplementation during mid-gestation
title_full_unstemmed Transcriptome profile in the skeletal muscle of cattle progeny as a function of maternal protein supplementation during mid-gestation
title_sort Transcriptome profile in the skeletal muscle of cattle progeny as a function of maternal protein supplementation during mid-gestation
dc.creator.fl_str_mv Carvalho, Elisa
Costa, Thaís C.
Sanglard, Leticia P.
Nascimento, Karolina B.
Moreno Meneses, Javier Andrés
Galvão, Matheus C.
Serão, Nick V.L.
Duarte, Marcio S.
Pies Gionbelli, Mateus
dc.contributor.author.none.fl_str_mv Carvalho, Elisa
Costa, Thaís C.
Sanglard, Leticia P.
Nascimento, Karolina B.
Moreno Meneses, Javier Andrés
Galvão, Matheus C.
Serão, Nick V.L.
Duarte, Marcio S.
Pies Gionbelli, Mateus
dc.subject.armarc.none.fl_str_mv Ganado bovino
topic Ganado bovino
Transcriptomas
Progenie
Gestación
dc.subject.agrovoc.none.fl_str_mv Transcriptomas
Progenie
Gestación
description This study aimed to evaluate the differentially expressed genes (DEG), their biological functions, and the main predictor genes in skeletal muscle of the offspring resulting from dams supplemented or not with protein during mid-gestation. From 100 to 200 days of gestation, 10 Tabapuã cows pregnant of males were assigned to one of two treatments: Control [(CON) - supply of basal diet achieving 5.5% of crude protein (CP); n = 6]; or Supplemented [(SUP) - basal diet plus a supplement with 40% of CP provided at the level of 3.5 g/kg of body weight; n = 4]. Muscle samples were collected from the progeny at 260d of postnatal age to RNA-seq, Gene Ontology (GO) enrichment, and Random Forest (RF) analyses. A total of 310 DEG (q-value < 0.05) were identified for the effect of diet, being 187 and 123 genes down-regulated and up-regulated in the SUP diet, respectively. Within the SUP up-regulated genes, were genes related to insulin receptor signaling pathway (LOC107131843) and apoptosis regulation (KRT18, KRT8 and KRT19). While the enriched GOs terms are implicated in mitochondrial metabolism through the overrepresentation of the GO terms microtubule polymerization or depolymerization (GO:0031109, GO:0046785) and positive regulation of cellular component biogenesis (GO:0044089). Moreover, RF analysis revealed predictor genes related to the skeletal muscle development, including genes that participates in the PI3K/Akt pathways (PITPNA), myoblast proliferation (ALKBH1) and differentiation (CDK5R1 and miR-206). In conclusion, maternal protein supplementation during mid-gestation positively affected the energy metabolism and favored the hypertrophic processes of the skeletal muscle of the offspring.
publishDate 2022
dc.date.accessioned.none.fl_str_mv 2022-08-22T18:46:56Z
dc.date.available.none.fl_str_mv 2022-08-22T18:46:56Z
dc.date.issued.none.fl_str_mv 2022
dc.type.spa.fl_str_mv Artículo de revista
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dc.identifier.citation.spa.fl_str_mv Carvalho, E. B., Costa, T. C., Sanglard, L. P., Nascimento, K. B., Meneses, J. A. M., Galvão, M. C., . . . Gionbelli, M. P. (2022). Transcriptome profile in the skeletal muscle of cattle progeny as a function of maternal protein supplementation during mid-gestation. Livestock Science, 263 doi:10.1016/j.livsci.2022.104995
dc.identifier.issn.spa.fl_str_mv 1871-1413
dc.identifier.uri.none.fl_str_mv https://repository.udca.edu.co/handle/11158/4821
dc.identifier.doi.none.fl_str_mv 10.1016/j.livsci.2022.104995
identifier_str_mv Carvalho, E. B., Costa, T. C., Sanglard, L. P., Nascimento, K. B., Meneses, J. A. M., Galvão, M. C., . . . Gionbelli, M. P. (2022). Transcriptome profile in the skeletal muscle of cattle progeny as a function of maternal protein supplementation during mid-gestation. Livestock Science, 263 doi:10.1016/j.livsci.2022.104995
1871-1413
10.1016/j.livsci.2022.104995
url https://repository.udca.edu.co/handle/11158/4821
dc.language.iso.spa.fl_str_mv eng
language eng
dc.relation.indexed.spa.fl_str_mv Agrosavia
dc.relation.citationendpage.spa.fl_str_mv 5
dc.relation.citationstartpage.spa.fl_str_mv 1
dc.relation.citationvolume.spa.fl_str_mv 263
dc.relation.ispartofjournal.spa.fl_str_mv Livestock Science
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spelling Carvalho, ElisaCosta, Thaís C.Sanglard, Leticia P.Nascimento, Karolina B.Moreno Meneses, Javier AndrésGalvão, Matheus C.Serão, Nick V.L.Duarte, Marcio S.Pies Gionbelli, Mateus2022-08-22T18:46:56Z2022-08-22T18:46:56Z2022Carvalho, E. B., Costa, T. C., Sanglard, L. P., Nascimento, K. B., Meneses, J. A. M., Galvão, M. C., . . . Gionbelli, M. P. (2022). Transcriptome profile in the skeletal muscle of cattle progeny as a function of maternal protein supplementation during mid-gestation. Livestock Science, 263 doi:10.1016/j.livsci.2022.1049951871-1413https://repository.udca.edu.co/handle/11158/482110.1016/j.livsci.2022.104995This study aimed to evaluate the differentially expressed genes (DEG), their biological functions, and the main predictor genes in skeletal muscle of the offspring resulting from dams supplemented or not with protein during mid-gestation. From 100 to 200 days of gestation, 10 Tabapuã cows pregnant of males were assigned to one of two treatments: Control [(CON) - supply of basal diet achieving 5.5% of crude protein (CP); n = 6]; or Supplemented [(SUP) - basal diet plus a supplement with 40% of CP provided at the level of 3.5 g/kg of body weight; n = 4]. Muscle samples were collected from the progeny at 260d of postnatal age to RNA-seq, Gene Ontology (GO) enrichment, and Random Forest (RF) analyses. A total of 310 DEG (q-value < 0.05) were identified for the effect of diet, being 187 and 123 genes down-regulated and up-regulated in the SUP diet, respectively. Within the SUP up-regulated genes, were genes related to insulin receptor signaling pathway (LOC107131843) and apoptosis regulation (KRT18, KRT8 and KRT19). While the enriched GOs terms are implicated in mitochondrial metabolism through the overrepresentation of the GO terms microtubule polymerization or depolymerization (GO:0031109, GO:0046785) and positive regulation of cellular component biogenesis (GO:0044089). Moreover, RF analysis revealed predictor genes related to the skeletal muscle development, including genes that participates in the PI3K/Akt pathways (PITPNA), myoblast proliferation (ALKBH1) and differentiation (CDK5R1 and miR-206). In conclusion, maternal protein supplementation during mid-gestation positively affected the energy metabolism and favored the hypertrophic processes of the skeletal muscle of the offspring.Incluye referencias bibliográficas.application/pdfenghttps://creativecommons.org/licenses/by-nc-sa/4.0/legalcode.eshttps://creativecommons.org/licenses/by-nc-sa/4.0/info:eu-repo/semantics/openAccessAtribución-NoComercial-CompartirIgual 4.0 Internacional (CC BY-NC-SA 4.0)http://purl.org/coar/access_right/c_abf2https://udca.elogim.com:2119/science/article/pii/S1871141322001743?via%3DihubTranscriptome profile in the skeletal muscle of cattle progeny as a function of maternal protein supplementation during mid-gestationArtículo de revistahttp://purl.org/coar/resource_type/c_2df8fbb1info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionTexthttp://purl.org/redcol/resource_type/ARThttp://purl.org/coar/version/c_970fb48d4fbd8a85Ganado 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texttext/plain33844https://repository.udca.edu.co/bitstreams/245160b4-7185-4705-8174-a8b6166addfe/downloadd88c13638de076d3975ca2e9cd24512bMD53THUMBNAILTranscriptome profile in the skeletal muscle of cattle progeny as a function of maternal protein supplementation during mid-gestation.pdf.jpgTranscriptome profile in the skeletal muscle of cattle progeny as a function of maternal protein supplementation during mid-gestation.pdf.jpgGenerated Thumbnailimage/jpeg14321https://repository.udca.edu.co/bitstreams/7481c9da-1601-42bb-a122-6a020bec1594/download201ae58646b918fe3ccc54f0498432dbMD5411158/4821oai:repository.udca.edu.co:11158/48212024-05-09 14:40:10.417https://creativecommons.org/licenses/by-nc-sa/4.0/https://creativecommons.org/licenses/by-nc-sa/4.0/legalcode.esopen.accesshttps://repository.udca.edu.coRepositorio - Universidad de Ciencias Aplicadas y Ambientales 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en las Obras Colectivas;
b.	Distribuir copias o fonogramas de las Obras, exhibirlas públicamente, ejecutarlas públicamente y/o ponerlas a disposición pública, incluyéndolas como incorporadas en Obras Colectivas, según corresponda;
c.	Distribuir copias de las Obras Derivadas que se generen, exhibirlas públicamente, ejecutarlas públicamente y/o ponerlas a disposición pública.
Los derechos mencionados anteriormente pueden ser ejercidos en todos los medios y formatos, actualmente conocidos o que se inventen en el futuro. Los derechos antes mencionados incluyen el derecho a realizar dichas modificaciones en la medida que sean técnicamente necesarias para ejercer los derechos en otro medio o formatos, pero de otra manera usted no está autorizado para realizar obras derivadas.Todos los derechos no otorgados expresamente por el Licenciante quedan por este medio reservados, incluyendo pero sin limitarse a aquellos que se mencionan en las secciones 4(d) y 4(e).
4. Restricciones.
La licencia otorgada en la anterior Sección 3 está expresamente sujeta y limitada por las siguientes restricciones:
a.	Usted puede distribuir, exhibir públicamente, ejecutar públicamente, o poner a disposición pública la Obra sólo bajo las condiciones de esta Licencia, y Usted debe incluir una copia de esta licencia o del Identificador Universal de Recursos de la misma con cada copia de la Obra que distribuya, exhiba públicamente, ejecute públicamente o ponga a disposición pública. No es posible ofrecer o imponer ninguna condición sobre la Obra que altere o limite las condiciones de esta Licencia o el ejercicio de los derechos de los destinatarios otorgados en este documento. No es posible sublicenciar la Obra. Usted debe mantener intactos todos los avisos que hagan referencia a esta Licencia y a la cláusula de limitación de garantías. Usted no puede distribuir, exhibir públicamente, ejecutar públicamente, o poner a disposición pública la Obra con alguna medida tecnológica que controle el acceso o la utilización de ella de una forma que sea inconsistente con las condiciones de esta Licencia. Lo anterior se aplica a la Obra incorporada a una Obra Colectiva, pero esto no exige que la Obra Colectiva aparte de la obra misma quede sujeta a las condiciones de esta Licencia. Si Usted crea una Obra Colectiva, previo aviso de cualquier Licenciante debe, en la medida de lo posible, eliminar de la Obra Colectiva cualquier referencia a dicho Licenciante o al Autor Original, según lo solicitado por el Licenciante y conforme lo exige la cláusula 4(c).
b.	Usted no puede ejercer ninguno de los derechos que le han sido otorgados en la Sección 3 precedente de modo que estén principalmente destinados o directamente dirigidos a conseguir un provecho comercial o una compensación monetaria privada. El intercambio de la Obra por otras obras protegidas por derechos de autor, ya sea a través de un sistema para compartir archivos digitales (digital file-sharing) o de cualquier otra manera no será considerado como estar destinado principalmente o dirigido directamente a conseguir un provecho comercial o una compensación monetaria privada, siempre que no se realice un pago mediante una compensación monetaria en relación con el intercambio de obras protegidas por el derecho de autor.
c.	Si usted distribuye, exhibe públicamente, ejecuta públicamente o ejecuta públicamente en forma digital la Obra o cualquier Obra Derivada u Obra Colectiva, Usted debe mantener intacta toda la información de derecho de autor de la Obra y proporcionar, de forma razonable según el medio o manera que Usted esté utilizando: (i) el nombre del Autor Original si está provisto (o seudónimo, si fuere aplicable), y/o (ii) el nombre de la parte o las partes que el Autor Original y/o el Licenciante hubieren designado para la atribución (v.g., un instituto patrocinador, editorial, publicación) en la información de los derechos de autor del Licenciante, términos de servicios o de otras formas razonables; el título de la Obra si está provisto; en la medida de lo razonablemente factible y, si está provisto, el Identificador Uniforme de Recursos (Uniform Resource Identifier) que el Licenciante especifica para ser asociado con la Obra, salvo que tal URI no se refiera a la nota sobre los derechos de autor o a la información sobre el licenciamiento de la Obra; y en el caso de una Obra Derivada, atribuir el crédito identificando el uso de la Obra en la Obra Derivada (v.g., "Traducción Francesa de la Obra del Autor Original," o "Guión Cinematográfico basado en la Obra original del Autor Original"). Tal crédito puede ser implementado de cualquier forma razonable; en el caso, sin embargo, de Obras Derivadas u Obras Colectivas, tal crédito aparecerá, como mínimo, donde aparece el crédito de cualquier otro autor comparable y de una manera, al menos, tan destacada como el crédito de otro autor comparable.
d.	Para evitar toda confusión, el Licenciante aclara que, cuando la obra es una composición musical:
i.	Regalías por interpretación y ejecución bajo licencias generales. El Licenciante se reserva el derecho exclusivo de autorizar la ejecución pública o la ejecución pública digital de la obra y de recolectar, sea individualmente o a través de una sociedad de gestión colectiva de derechos de autor y derechos conexos (por ejemplo, SAYCO), las regalías por la ejecución pública o por la ejecución pública digital de la obra (por ejemplo Webcast) licenciada bajo licencias generales, si la interpretación o ejecución de la obra está primordialmente orientada por o dirigida a la obtención de una ventaja comercial o una compensación monetaria privada.
ii.	Regalías por Fonogramas. El Licenciante se reserva el derecho exclusivo de recolectar, individualmente o a través de una sociedad de gestión colectiva de derechos de autor y derechos conexos (por ejemplo, SAYCO), una agencia de derechos musicales o algún agente designado, las regalías por cualquier fonograma que Usted cree a partir de la obra (“versión cover”) y distribuya, en los términos del régimen de derechos de autor, si la creación o distribución de esa versión cover está primordialmente destinada o dirigida a obtener una ventaja comercial o una compensación monetaria privada.
e.	Gestión de Derechos de Autor sobre Interpretaciones y Ejecuciones Digitales (WebCasting). Para evitar toda confusión, el Licenciante aclara que, cuando la obra sea un fonograma, el Licenciante se reserva el derecho exclusivo de autorizar la ejecución pública digital de la obra (por ejemplo, webcast) y de recolectar, individualmente o a través de una sociedad de gestión colectiva de derechos de autor y derechos conexos (por ejemplo, Acinpro), las regalías por la ejecución pública digital de la obra (por ejemplo, webcast), sujeta a las disposiciones aplicables del régimen de Derecho de Autor, si esta ejecución pública digital está primordialmente dirigida a obtener una ventaja comercial o una compensación monetaria privada.
5. Representaciones, Garantías y Limitaciones de Responsabilidad.
A MENOS QUE LAS PARTES LO ACORDARAN DE OTRA FORMA POR ESCRITO, EL LICENCIANTE OFRECE LA OBRA (EN EL ESTADO EN EL QUE SE ENCUENTRA) “TAL CUAL”, SIN BRINDAR GARANTÍAS DE CLASE ALGUNA RESPECTO DE LA OBRA, YA SEA EXPRESA, IMPLÍCITA, LEGAL O CUALQUIERA OTRA, INCLUYENDO, SIN LIMITARSE A ELLAS, GARANTÍAS DE TITULARIDAD, COMERCIABILIDAD, ADAPTABILIDAD O ADECUACIÓN A PROPÓSITO DETERMINADO, AUSENCIA DE INFRACCIÓN, DE AUSENCIA DE DEFECTOS LATENTES O DE OTRO TIPO, O LA PRESENCIA O AUSENCIA DE ERRORES, SEAN O NO DESCUBRIBLES (PUEDAN O NO SER ESTOS DESCUBIERTOS). ALGUNAS JURISDICCIONES NO PERMITEN LA EXCLUSIÓN DE GARANTÍAS IMPLÍCITAS, EN CUYO CASO ESTA EXCLUSIÓN PUEDE NO APLICARSE A USTED.
6. Limitación de responsabilidad.
A MENOS QUE LO EXIJA EXPRESAMENTE LA LEY APLICABLE, EL LICENCIANTE NO SERÁ RESPONSABLE ANTE USTED POR DAÑO ALGUNO, SEA POR RESPONSABILIDAD EXTRACONTRACTUAL, PRECONTRACTUAL O CONTRACTUAL, OBJETIVA O SUBJETIVA, SE TRATE DE DAÑOS MORALES O PATRIMONIALES, DIRECTOS O INDIRECTOS, PREVISTOS O IMPREVISTOS PRODUCIDOS POR EL USO DE ESTA LICENCIA O DE LA OBRA, AUN CUANDO EL LICENCIANTE HAYA SIDO ADVERTIDO DE LA POSIBILIDAD DE DICHOS DAÑOS. ALGUNAS LEYES NO PERMITEN LA EXCLUSIÓN DE CIERTA RESPONSABILIDAD, EN CUYO CASO ESTA EXCLUSIÓN PUEDE NO APLICARSE A USTED.
7. Término.
a.	Esta Licencia y los derechos otorgados en virtud de ella terminarán automáticamente si Usted infringe alguna condición establecida en ella. Sin embargo, los individuos o entidades que han recibido Obras Derivadas o Colectivas de Usted de conformidad con esta Licencia, no verán terminadas sus licencias, siempre que estos individuos o entidades sigan cumpliendo íntegramente las condiciones de estas licencias. Las Secciones 1, 2, 5, 6, 7, y 8 subsistirán a cualquier terminación de esta Licencia.
b.	Sujeta a las condiciones y términos anteriores, la licencia otorgada aquí es perpetua (durante el período de vigencia de los derechos de autor de la obra). No obstante lo anterior, el Licenciante se reserva el derecho a publicar y/o estrenar la Obra bajo condiciones de licencia diferentes o a dejar de distribuirla en los términos de esta Licencia en cualquier momento; en el entendido, sin embargo, que esa elección no servirá para revocar esta licencia o que deba ser otorgada , bajo los términos de esta licencia), y esta licencia continuará en pleno vigor y efecto a menos que sea terminada como se expresa atrás. La Licencia revocada continuará siendo plenamente vigente y efectiva si no se le da término en las condiciones indicadas anteriormente.
8. Varios.
a.	Cada vez que Usted distribuya o ponga a disposición pública la Obra o una Obra Colectiva, el Licenciante ofrecerá al destinatario una licencia en los mismos términos y condiciones que la licencia otorgada a Usted bajo esta Licencia.
b.	Si alguna disposición de esta Licencia resulta invalidada o no exigible, según la legislación vigente, esto no afectará ni la validez ni la aplicabilidad del resto de condiciones de esta Licencia y, sin acción adicional por parte de los sujetos de este acuerdo, aquélla se entenderá reformada lo mínimo necesario para hacer que dicha disposición sea válida y exigible.
c.	Ningún término o disposición de esta Licencia se estimará renunciada y ninguna violación de ella será consentida a menos que esa renuncia o consentimiento sea otorgado por escrito y firmado por la parte que renuncie o consienta.
d.	Esta Licencia refleja el acuerdo pleno entre las partes respecto a la Obra aquí licenciada. No hay arreglos, acuerdos o declaraciones respecto a la Obra que no estén especificados en este documento. El Licenciante no se verá limitado por ninguna disposición adicional que pueda surgir en alguna comunicación emanada de Usted. Esta Licencia no puede ser modificada sin el consentimiento mutuo por escrito del Licenciante y Usted.

