Analysing the Expression of Eight Clock Genes in Five Tissues From Fasting and Fed Sows

In a previous study, we observed that circadian clock genes are differentially expressed in the skeletal muscle of fasting and fed sows. The goal of the current work was to investigate if these genes are also differentially expressed in tissues containing the central (hypothalamus) and peripheral (d...

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Main Authors: Tainã Figueiredo Cardoso, Raquel Quintanilla, Anna Castelló, Emilio Mármol-Sánchez, Maria Ballester, Jordi Jordana, Marcel Amills
Format: Article
Language:English
Published: Frontiers Media S.A. 2018-10-01
Series:Frontiers in Genetics
Subjects:
Online Access:https://www.frontiersin.org/article/10.3389/fgene.2018.00475/full
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author Tainã Figueiredo Cardoso
Tainã Figueiredo Cardoso
Raquel Quintanilla
Anna Castelló
Anna Castelló
Emilio Mármol-Sánchez
Maria Ballester
Jordi Jordana
Marcel Amills
Marcel Amills
author_facet Tainã Figueiredo Cardoso
Tainã Figueiredo Cardoso
Raquel Quintanilla
Anna Castelló
Anna Castelló
Emilio Mármol-Sánchez
Maria Ballester
Jordi Jordana
Marcel Amills
Marcel Amills
author_sort Tainã Figueiredo Cardoso
collection DOAJ
description In a previous study, we observed that circadian clock genes are differentially expressed in the skeletal muscle of fasting and fed sows. The goal of the current work was to investigate if these genes are also differentially expressed in tissues containing the central (hypothalamus) and peripheral (duodenum, dorsal fat, muscle, and liver) clocks. As animal material, we used 12 sows that fasted 12 h before slaughtering (T0) and 12 sows that were fed ad libitum 7 h prior slaughtering (T2). Tissue samples were collected immediately after slaughter and total RNA was subsequently extracted. The expression of the ARNTL, BHLHE40, CRY2, NPAS2, NR1D1, PER1, PER2, and SIK1 genes was measured by quantitative reverse transcription PCR. The numbers of clock genes showing differential expression before and after feeding varied depending on the tissue i.e., four in dorsal fat and duodenum, six in skeletal muscle, and seven in the liver. In contrast, none of the eight analysed genes displayed a significant differential expression in hypothalamus, the tissue where the central clock resides. This result supports that the differential expression of clock genes in the four tissues mentioned above is probably induced by nutrition and not by the central clock entrained by light. Moreover, we have observed that the NPAS2 and ARNTL genes display positive log2(FC) values in the five tissues under analysis, whilst the CRY2, PER1 (except dorsal fat) and PER2 (except hypothalamus) genes generally show negative log2(FC) values. Such result might be explained by the existence of a negative feedback loop between the ARNTL/NPAS2 and CRY/PER genes. Collectively, these results support that nutrition plays an important role in modulating the timing of porcine peripheral circadian clocks. Such regulation could be essential for coordinating the subsequent metabolic response to nutrient supply.
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spelling doaj.art-09ae1ec7b2c14ea9a1441d1576ca43c92022-12-21T20:11:29ZengFrontiers Media S.A.Frontiers in Genetics1664-80212018-10-01910.3389/fgene.2018.00475371614Analysing the Expression of Eight Clock Genes in Five Tissues From Fasting and Fed SowsTainã Figueiredo Cardoso0Tainã Figueiredo Cardoso1Raquel Quintanilla2Anna Castelló3Anna Castelló4Emilio Mármol-Sánchez5Maria Ballester6Jordi Jordana7Marcel Amills8Marcel Amills9Department of Animal Genetics, Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB, Campus de la Universitat Autònoma de Barcelona, Bellaterra, SpainCAPES Foundation, Ministry of Education of Brazil, Brasilia, BrazilAnimal Breeding and Genetics Programme, The Institute for Research and Technology in Food and Agriculture (IRTA), Barcelona, SpainDepartment of Animal Genetics, Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB, Campus de la Universitat Autònoma de Barcelona, Bellaterra, SpainDepartament de Ciència Animal i dels Aliments, Universitat Autònoma de Barcelona, Bellaterra, SpainDepartment of Animal Genetics, Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB, Campus de la Universitat Autònoma de Barcelona, Bellaterra, SpainAnimal Breeding and Genetics Programme, The Institute for Research and Technology in Food and Agriculture (IRTA), Barcelona, SpainDepartament de Ciència Animal i dels Aliments, Universitat Autònoma de Barcelona, Bellaterra, SpainDepartment of Animal Genetics, Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB, Campus de la Universitat Autònoma de Barcelona, Bellaterra, SpainDepartament de Ciència Animal i dels Aliments, Universitat Autònoma de Barcelona, Bellaterra, SpainIn a previous study, we observed that circadian clock genes are differentially expressed in the skeletal muscle of fasting and fed sows. The goal of the current work was to investigate if these genes are also differentially expressed in tissues containing the central (hypothalamus) and peripheral (duodenum, dorsal fat, muscle, and liver) clocks. As animal material, we used 12 sows that fasted 12 h before slaughtering (T0) and 12 sows that were fed ad libitum 7 h prior slaughtering (T2). Tissue samples were collected immediately after slaughter and total RNA was subsequently extracted. The expression of the ARNTL, BHLHE40, CRY2, NPAS2, NR1D1, PER1, PER2, and SIK1 genes was measured by quantitative reverse transcription PCR. The numbers of clock genes showing differential expression before and after feeding varied depending on the tissue i.e., four in dorsal fat and duodenum, six in skeletal muscle, and seven in the liver. In contrast, none of the eight analysed genes displayed a significant differential expression in hypothalamus, the tissue where the central clock resides. This result supports that the differential expression of clock genes in the four tissues mentioned above is probably induced by nutrition and not by the central clock entrained by light. Moreover, we have observed that the NPAS2 and ARNTL genes display positive log2(FC) values in the five tissues under analysis, whilst the CRY2, PER1 (except dorsal fat) and PER2 (except hypothalamus) genes generally show negative log2(FC) values. Such result might be explained by the existence of a negative feedback loop between the ARNTL/NPAS2 and CRY/PER genes. Collectively, these results support that nutrition plays an important role in modulating the timing of porcine peripheral circadian clocks. Such regulation could be essential for coordinating the subsequent metabolic response to nutrient supply.https://www.frontiersin.org/article/10.3389/fgene.2018.00475/fullclock genesfood ingestionnutritionRT-qPCRpig
spellingShingle Tainã Figueiredo Cardoso
Tainã Figueiredo Cardoso
Raquel Quintanilla
Anna Castelló
Anna Castelló
Emilio Mármol-Sánchez
Maria Ballester
Jordi Jordana
Marcel Amills
Marcel Amills
Analysing the Expression of Eight Clock Genes in Five Tissues From Fasting and Fed Sows
Frontiers in Genetics
clock genes
food ingestion
nutrition
RT-qPCR
pig
title Analysing the Expression of Eight Clock Genes in Five Tissues From Fasting and Fed Sows
title_full Analysing the Expression of Eight Clock Genes in Five Tissues From Fasting and Fed Sows
title_fullStr Analysing the Expression of Eight Clock Genes in Five Tissues From Fasting and Fed Sows
title_full_unstemmed Analysing the Expression of Eight Clock Genes in Five Tissues From Fasting and Fed Sows
title_short Analysing the Expression of Eight Clock Genes in Five Tissues From Fasting and Fed Sows
title_sort analysing the expression of eight clock genes in five tissues from fasting and fed sows
topic clock genes
food ingestion
nutrition
RT-qPCR
pig
url https://www.frontiersin.org/article/10.3389/fgene.2018.00475/full
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