Search

Pig industry is facing new challenges that make necessary to reorient breeding programs to produce more robust and resilient pig populations. The aim of the present work was to study the genetic determinism of lymphocyte subpopulations in the peripheral blood of pigs and identify genomic regions and biomarkers associated to them. For this purpose, we stained peripheral blood mononuclear cells to measure ten immune-cell-related traits including the relative abundance of different populations of lymphocytes, the proportions of CD4+ T cells and CD8+ T cells, and the ratio of CD4+/CD8+ T cells from 391 healthy Duroc piglets aged 8 weeks. Medium to high heritabilities were observed for the ten immune-cell-related traits and significant genetic correlations were obtained between the proportion of some lymphocytes populations. A genome-wide association study pointed out 32 SNPs located at four chromosomal regions on pig chromosomes SSC3, SSC5, SSC8, and SSCX as significantly associated to T-helper cells, memory T-helper cells and γδ T cells. Several genes previously identified in human association studies for the same or related traits were located in the associated regions, and were proposed as candidate genes to explain the variation of T cell populations such as CD4, CD8A, CD8B, KLRC2, RMND5A and VPS24. The transcriptome analysis of whole blood samples from animals with extreme proportions of γδ T, T-helper and memory T-helper cells identified differentially expressed genes (CAPG, TCF7L1, KLRD1 and CD4) located into the associated regions. In addition, differentially expressed genes specific of different T cells subpopulations were identified such as SOX13 and WC1 genes for γδ T cells. Our results enhance the knowledge about the genetic control of lymphocyte traits that could be considered to optimize the induction of immune responses to vaccines against pathogens. Furthermore, they open the possibility of applying effective selection programs for improving immunocompetence in pigs and support the use of the pig as a very reliable human biomedical model.

Correcciones en la sección de Agradecimientos del artículo original, donde estaba incompleta., Correction to: Scientific Reports https://doi.org/10.1038/srep31803, published online 18 August 2016., This work was funded by the Ministerio de Economía y Competitividad (MINECO AGL2011-29821-C02 and AGL2014-56369-C2) and the Fondo Europeo de Desarrollo Regional (FEDER). A. Puig-Oliveras was funded by a “Personal Investigador en Formació” (PIF) grant from Universitat Autònoma de Barcelona (458-01-1/2011). M. Revilla was funded by a Formació i Contractació de Personal Investigador Novell (FI-DGR) Ph.D Grant from the Generalitat de Catalunya (ECO/1639/2013). M. Ballester is financially supported by a Ramon y Cajal contract (RYC-2013-12573) from the Spanish Ministry of Economy and Competitiveness.

Feed efficiency (FE) is one of the most economically and environmentally relevant traits in the animal production sector. The objective of this study was to gain knowledge about the genetic control of FE in rabbits. To this end, GWASs were conducted for individual growth under two feeding regimes (full feeding and restricted) and FE traits collected from cage groups, using 114 604 autosome SNPs segregating in 438 rabbits. Two different models were implemented: (1) an animal model with a linear regression on each SNP allele for growth trait; and (2) a two-trait animal model, jointly fitting the performance trait and each SNP allele content, for FE traits. This last modeling strategy is a new tool applied to GWAS and allows information to be considered from non-genotyped individuals whose contribution is relevant in the group average traits. A total of 189 SNPs in 17 chromosomal regions were declared to be significantly associated with any of the five analyzed traits at a chromosome-wide level. In 12 of these regions, 20 candidate genes were proposed to explain the variation of the analyzed traits, including genes such as FTO, NDUFAF6 and CEBPA previously associated with growth and FE traits in monogastric species. Candidate genes associated with behavioral patterns were also identified. Overall, our results can be considered as the foundation for future functional research to unravel the actual causal mutations regulating growth and FE in rabbits.

Intramuscular fat content and its fatty acid composition affect porcine meat quality and its nutritional value. The present work aimed to identify genomic variants regulating the expression in the porcine muscle (Longissimus dorsi) of 45 candidate genes for lipid metabolism and fatty acid composition in three experimental backcrosses based on the Iberian breed. Expression genome-wide association studies (eGWAS) were performed between the muscle gene expression values, measured by real-time quantitative PCR, and the genotypes of 38,426 SNPs distributed along all chromosomes. The eGWAS identified 186 eSNPs located in ten Sus scrofa regions and associated with the expression of ACSM5, ACSS2, ATF3, DGAT2, FOS and IGF2 (FDR, This work was supported by the Spanish Ministerio de Economía y Competitividad (MINECO) and the Fondo Europeo de Desarrollo Regional (FEDER) with project references: AGL2014–56369-C2 and AGL2017–82641-R. L. Criado-Mesas was funded with a FPI grant from the AGL2014–56369-C2 project. M. Ballester was financially supported by a “Ramón y Cajal” contract (RYC-2013–12573) from the Spanish Ministerio de Economía y Competitividad. D. Crespo-Piazuelo was funded by a “Formació i Contractació de Personal Investigador Novell” (FI-DGR) Ph.D grant from the Generalitat de Catalunya (ECO/1788/2014). We acknowledge the support of the Spanish Ministerio de Economía y Competitividad for the “Severo Ochoa Programme for Centres of Excellence in R&D” 2016–2019 (SEV-2015–0533) to the Centre for Research in Agricultural Genomics and the CERCA Programme / Generalitat de Catalunya.

[Background]: The molecular basis of compensatory growth in monogastric animals has not yet been fully explored. Herewith, in this study we aim to determine changes in the pig skeletal muscle transcriptome profile during compensatory growth following a feed restriction period. A RNA-Seq experiment was performed with a total of 24 females belonging to a Duroc commercial line. Half of the animals received either a restricted (RE) or ad libitum (AL) diet during the first fattening period (60–125 d of age). After that, all gilts were fed ad libitum for a further ~30 d until the age of ~155 d, when animals were slaughtered and samples of gluteus medius muscle were harvested to perform RNA-Seq analyses and intramuscular fat content determination., [Results]: During the period following food restriction, RE animals re-fed ad libitum displayed compensatory growth, showed better feed conversion rate and tended to deposit more subcutaneous fat than AL fed animals. Animals were slaughtered in the phase of accelerated growth, when RE animals had not completely compensated the performance of AL group, showing lower live and carcass weights. At intramuscular level, RE gilts showed a higher content of polyunsaturated fatty acids during the compensatory growth phase. The comparison of RE and AL expression profiles allowed the identification of 86 (ǀlog2Fold-Changeǀ > 1, padj, [Conclusions]: Animals re-fed after a restriction period exhibited a less oxidative metabolic profile and catabolic processes in muscle than animals fed ad libitum. The downregulation of autophagy observed in the skeletal muscle of pigs undergoing compensatory growth may constitute a mechanism to increase muscle mass thus ensuring an accelerated growth rate. These results reveal that the downregulation of AMPK Signaling plays an important role in compensatory growth in pigs., The study was funded by grants AGL2013–48742-C2–1-R and AGL2013–48742-C2–2-R, awarded by the Spanish Ministry of Economy and Competitivity (MINECO). We also acknowledge the support of the Spanish Ministry of Economy and Competitivity for the Center of Excellence Severo Ochoa 2016–2019 (SEV-2015-0533) grant awarded to the Centre for Research in Agricultural Genomics (CRAG). Maria Ballester is financially supported by a Ramon y Cajal contract (RYC-2013-12573) from the Spanish Ministry of Economy and Competitiveness. Tainã Figueiredo Cardoso was funded with a fellowship from the CAPES Foundation-Coordination of Improvement of Higher Education, Ministry of Education of the Federal Government of Brazil. Rayner Gonzalez-Prendes was funded with a FPU Ph.D. grant from the Spanish Ministry of Education (FPU12/00860). Thanks also to the CERCA Programme of the Generalitat de Catalunya.

Combining animal production performances and health traits is a main issue in livestock farming. Blood is emerging as a source of biological information linked to the health and physiological state of each individual. Furthermore, it is an interesting sur rogate tissue for animal phenotyping because its sampling is almost non-invasive and reproducible. We have already shown that several immune parameters are heritable and that the blood transcriptome is informative to quantify some of them. In this study, our aim was to identify genetic markers associated with gene expression variations in the blood, based on an expression genome-wide association study (eGWAS) using 243 Large White pigs at 60 days of age. Each pig was genotyped with the Illumina iSelect 60K Chip and the blood transcriptome analysis was performed by using a custom gene expression 8X60K microarray (Agilent Technologies). 4,591 significant associations between one SNP and one probe were found, including 62% cis-associations (the associated SNP is in a window of 2Mb surrounding the probe). Among the 3,419 probes with at least one associated SNP (eQTL), 70% had a cis-eQTL and 89% could be assigned to a porcine gene. The eQTLs were found to be linked with the expression of one to 51 genes. The genomic regions surrounding the eQTLs correspond to candidate regions to further study thegenetic architecture of immunity trait variations in pigs, and contribute to paving the way towards a qualification of the individual immune capacity for tran slational research in precision pig farming.

Estudi realitzat a partir d’una estada al Institut National de la Recherche Agronomique (INRA), a França, entre 2006 i 2008. En el ultims anys, estudis realitzats en diferents tipus cel•lulars han pogut determinar l’importància de l’organització nuclear en el control i regulació gènica. S’han realizat diferents experiments per tal de determinar si la posició dels gens de les proteïnes làcties en el nucli interfàsic de cel•lules epitelials mamaries és important per regular la seva expressió. Els gens de les proteïnes de la llet s’expressen a la glàndula mamaria durant la lactació en resposta a les hormones lactogèniques (majoritàriament prolactina i glucocorticoids). Mitjançant la tècnica de FISH (fluorescent in situ hibridization) en 3D s’ha caracteritzat la localització nuclear del gens WAP (whey acidic protein) i les caseïnes en cèl•lules epitelials mamaries de ratolí (HC11) cultivades en l’absència i presencia d’hormones lactogèniques. En absència d’hormones, els dos gens estan distribuïts dins del nucli de forma no aleatòria, el gen WAP es troba localitzat en l’interior del nucli, mentre que les caseïnes es troben localitzades prop de la perifèria nuclear. L’estimulació hormonal indueix un canvi significatiu en la distància dels dos gens a la perifèria nuclear. Així mateix, la posició del locus de la caseïna en relació al seu territori cromosòmic (CT) 5 està correlacionada amb la inducció hormonal i per tant amb la seva activació transcripcional, mentre que la posició del gen WAP amb relació al seu CT11 sembla més determinada pel context cromosòmic del gen. Per últim, no s’han trobat diferencies en la localització dels gens en relació a l'heterocromatina del centròmer, descrit com a compartiment repressiu, entre les cèl•lules estimulades amb hormones i les que no. En els dos casos s’ha trobat un gran percentatge de gens que no estan associats als centròmers.

Els programes de fecundació in vitro utilitzen sovint una hormona que secreta la glàndula pituïtària. Obtenir aquestes hormones resulta molt costós. Investigadors de la UAB estan treballant per tal d'obtenir més eficaçment aquesta hormona utilitzant ratolins transgènics per testar in vivo les construccions transgèniques. Los programas de fecundación in vitro utilizan a menudo una hormonaque secreta la glándula pituitaria. Obtener estas hormonas resulta muycostoso. Investigadores de la UAB están trabajando para conseguir máseficazmente esta hormona utilizando ratones transgénicos para testar invivo las construcciones transgénicas.

El objetivo final de nuestro trabajo es la obtención de proteínas recombinantes de interés farmacológico en la leche de animales transgénicos de producción. En nuestro grupo, previamente a este trabajo, se han llevado a cabo distintos proyectos con el fin de estudiar las regiones reguladoras del gen de la b-lactoglobulina (bLG) caprina y obtener así un cassette de expresión que pueda dirigir la expresión del transgén de forma específica y eficaz a la glándula mamaria de los animales transgénicos. Todas las construcciones han sido testadas previamente en el ratón al ser un modelo animal más barato ya que presenta un menor coste de mantenimiento y su ciclo reproductivo e intervalo generacional es más corto con respecto a los animales de producción. En el presente trabajo se ha utilizado un cassette de expresión que contiene 410 pb de región promotora proximal y 2,5 kb de región 3' flanqueante del gen de la bLG caprina para dirigir la expresión de las dos subunidades (a y b) de la Hormona Estimulante del Folículo (FSH) humana a la glándula mamaria de ratones transgénicos. Se han obtenido 4 líneas transgénicas que son capaces de expresar los dos transgenes en su glándula mamaria. Por otra parte, en la obtención de ratones transgénicos para el transgén de la bLG caprina, con el fin de testar las regiones reguladoras del gen, se observó un fenotipo conspicuo en los ratones homocigotos de la línea tg56; en el presente trabajo se ha caracterizado el lugar de integración de este transgén, el cual provocó una mutación por inserción en el gen de la Fosfolipasa C-b1 (PLC-b1) del ratón.Asimismo, se ha desarrollado una nuevo protocolo basado en la PCR cuantitativa a tiempo real para determinar el número de copias integradas del transgén en las distintas líneas transgénicas. Este método puede ser utilizado en todas aquellas líneas transgénicas que contengan cassettes de expresión pertenecientes a genes de especies rumiantes.Por último se han identificado y caracterizado un total de 15 polimorfismos en el gen de la bLG caprina, 9 de los cuales se encuentran localizados en la región promotora proximal, con el fin de encontrar mutaciones en las regiones reguladoras que puedan ser importantes en la expresión del gen., The final aim of this project is to produce pharmaceutical recombinant proteins in milk of farm animals. Previous works of our group have been performed to study the caprine b-lactoglobulin (bLG) regulatory sequences resulting in a expression cassette that is able to target the expression of the transgene to the mammary gland of transgenic mice in an integration-site dependent manner, independently of the number of copies in the array. All the constructions have been previously tested in a mouse model due to their short generation intervals and ease of genetic manipulation being cheaper than farm animals.In the present work, a goat bLG expression cassette which contained 410 bp of proximal promoter region and 2.5 kb of 3' flanking region has been used to target the expression of the two subunits (a and b) of human Follicle Stimulating Hormone (hFSH) to the mammary gland of transgenic mice. Four established transgenic lines that express both hFSH transgenes (a and b) to their mammary gland have been obtained.On the other hand, homozygous mice from one (tg56) of the different transgenic lines, that have been previously obtained for the goat bLG gene to study the regulatory sequences, displayed a distinct phenotype. Here, we present the identification and characterization of the transgene-induced insertional mutation at the PLC-b1 locus (PLC-b1bLG) in line tg56 of bLG transgenic mice.Furthermore, a rapid and accurate real-time quantitative PCR-based system to determine transgene copy number in transgenic animals has been developed. This method can be used directly for the analysis of transgenic mice for transgenes with different ruminant sequences without the requirement of control sample previously quantified by blotting techniques.Finally, fifteen polymorphisms, nine in the promoter region, of the caprine bLG gene have been identified and characterized. The main aim of this study has been identified mutations in the proximal promoter region that could be important in the expression of the gene.

La carne de cerdo es una de las carnes más consumidas en el mundo, cuyo valor se ve afectado por su calidad y las preferencias del consumidor. La composición de los ácidos grasos (AGs) en músculo y tejido adiposo modifica la calidad de la carne. Del mismo modo, la microbiota intestinal, a través de la producción de metabolitos como los ácidos grasos volátiles, puede también afectar su calidad. Sin embargo, la relación entre el genoma del cerdo y su microbiota intestinal no está bien estudiada. En la presente tesis se han realizado una serie de trabajos con el fin de profundizar en los mecanismos genéticos implicados en la determinación de la composición de los AGs. Además, se ha estudiado la composición de la microbiota a lo largo del intestino y su interacción con el genoma porcino. Se realizaron estudios de asociación del genoma completo (GWAS) entre 38.424 Single Nucleotide Polymorphisms (SNPs) y 60 caracteres fenotípicos relacionados con la composición de los AGs en músculo y grasa dorsal de 441 cerdos pertenecientes a tres retrocruces de la población experimental IBMAP: BC1_LD (25% Ibérico y 75% Landrace), BC1_PI (25% Ibérico y 75% Pietrain), y BC1_DU (25% Ibérico y 75% Duroc). El GWAS reveló nueve regiones del genoma porcino asociadas con doce caracteres de la grasa dorsal y seis regiones asociadas con seis medidas de la grasa intramuscular. Dentro de estas regiones, se identificaron 50 genes como candidatos funcionales a explicar la variación de estos caracteres. Los genes más relevantes fueron ELOVL3, ELOVL6, ELOVL7, FADS2, FASN y SCD. Además, el polimorfismo ELOVL6:c.‑394G>A fue el más asociado con los porcentajes de C14:0, C16:0, y C16:1(n-7) en grasa dorsal. Para estudiar otras variantes genéticas aparte de los SNPs, se detectaron 1.928.746 indels con tres programas (Dindel, SAMtools mpileup, y GATK) mediante los datos de secuenciación del genoma completo de siete fundadores (dos machos Ibéricos y cinco hembras Landrace) del material IBMAP. Se genotiparon diez indels localizados en genes relacionados con el metabolismo lipídico en los 441 cerdos de los tres retrocruces, encontrándose a distintas frecuencias alélicas. En la grasa intramuscular, el indel C1QTNF12:c.557_559delCCG presentó una asociación significativa con el porcentaje de ácido eicosadienoico (C20:2(n-6)). Para describir la composición de la microbiota a lo largo del intestino, se recogió el contenido luminal de cinco regiones (duodeno, yeyuno, íleo, colon proximal y distal) de trece cerdos Ibéricos. Posteriormente, mediante el método de amplificación y secuenciación del gen 16S rRNA, se identificaron 1.669 operational taxonomic units (OTUs) agrupados en 179 géneros, siendo los más abundantes Lactobacillus, Clostridium y Prevotella. Las muestras de colon eran más ricas en especies y se parecían más entre cerdos que las muestras del intestino delgado. Además, las predicciones funcionales del metagenoma a lo largo del intestino mostraron que sus rutas energéticas eran distintas. Finalmente, se estudió la asociación entre el genoma del cerdo y su microbiota intestinal. Se obtuvo la composición de la microbiota del recto de 285 cerdos Ibérico × Duroc mediante la amplificación y secuenciación del gen del 16S rRNA, identificándose un total de 1.257 OTUs agrupados en 101 géneros y 18 filos, siendo los filos más abundantes Firmicutes y Bacteroidetes. El GWAS reveló 17 regiones del genoma porcino asociadas con la abundancia relativa de los géneros Akkermansia, CF231, Phascolarctobacterium, Prevotella, SMB53 y Streptococcus. Dentro de estas regiones, se identificaron 38 genes como candidatos a modular la composición de la microbiota intestinal por su relación con el sistema inmunitario y el metabolismo de los mucopolisacáridos y los ácidos biliares. Pork is one of the most consumed meats worldwide and it is subjected to consumer’s preferences. Meat quality is affected by fatty acid (FA) composition in muscle and adipose tissues. Gut microbiota composition can also affect meat quality through the production of metabolites such as short-chain fatty acids. However, the relationship between pig genome and gut microbiota is not fully understood. In the current thesis, several studies have been performed to improve our knowledge about the genetic determinism of FA composition. In addition, the composition of the microbiota along the pig gut and its interaction with the host genome has been also analysed. Genome-wide association studies (GWAS) were performed among 38,424 single nucleotide polymorphisms (SNPs) and 60 phenotypic traits related to FA composition in backfat and muscle. This analysis was performed in 441 pigs from three different backcrosses: BC1_LD (25% Iberian and 75% Landrace), BC1_PI (25% Iberian and 75% Pietrain), and BC1_DU (25% Iberian and 75% Duroc) belonging to the IBMAP experimental population. Nine regions of the pig genome were associated with twelve backfat traits, while six regions were associated with six intramuscular fat (IMF) traits. A total of 50 candidate genes were proposed to explain the variation in these traits. The most promising candidate genes were ELOVL3, ELOVL6, ELOVL7, FADS2, FASN and SCD. Furthermore, ELOVL6:c.‑394G>A was the most associated SNP with the percentages of C14:0, C16:0, and C16:1(n-7) in backfat. With the aim of detecting other variants apart from SNPs, we performed an indel detection with the whole genome sequencing data from seven founders (two Iberian boars and five Landrace sows) of the IBMAP pigs. A total of 1,928,746 indels were found in common among the three programs used (Dindel, SAMtools mpileup, and GATK). Ten indels inside genes related with lipid metabolism (ASPH, C1QTNF12, CAPN9, CCR7, CRP, GZMA, JMJD1C, LYST, PEX19 and SAMD4B) were genotyped in pigs belonging to the three IBMAP backcrosses, obtaining different allelic frequencies. The C1QTNF12:c.557_559delCCG indel was associated with the percentage of eicosadienoic acid (C20:2(n-6)) in IMF. To describe the microbiota composition along the pig gut, luminal contents of five gut sections (duodenum, jejunum, ileum, and proximal and distal colon) were collected in thirteen Iberian pigs. A total of 1,669 operational taxonomic units (OTUs) grouped in 179 genera were found using the 16S rRNA gene sequencing method. Lactobacillus, Clostridium and Prevotella were the three most abundant genera. Colon samples were more similar among pigs and richer in species than small intestine samples were. The metagenome predictions showed that the energy pathways were different along gut sections. Finally, to reveal the association between host genome and gut microbiota in pigs, the microbiota composition of the rectum of 285 Iberian × Duroc pigs was obtained using the 16S rRNA gene sequencing method, finding 1,257 OTUs distributed in 101 genera and 18 phyla. Firmicutes and Bacteroidetes were the most abundant phyla. GWAS identified 17 genomic regions of the pig genome associated with the relative abundance of six genera (Akkermansia, CF231, Phascolarctobacterium, Prevotella, SMB53 and Streptococcus). A total of 38 candidate genes, related with the host defence system and the metabolism of mucopolysaccharides and bile acids, were suggested to be modulators of the gut microbiota composition.

El cerdo es una de las principales fuentes de carne consumida por el hombre y las preferencias de los consumidores hacia productos de alta calidad han aumentado durante los últimos años. Por lo tanto, conocer los mecanismos moleculares que afectan a la producción y a la calidad de esta carne ayudaría a la selección de estos caracteres. La calidad de la carne está determinada en gran medida por la composición de los ácidos grasos (AG) y la comprensión de los procesos moleculares subyacentes a éstos son el objetivo general de esta tesis. En este trabajo, se han identificado QTLs en el cromosoma 8 porcino (SSC8) para la composición de AG en grasa dorsal (GD) y se han identificado dos regiones cromosómicas con SNPs asociados, localizadas a 93 y 119 Mb. Las señales estadísticamente más significativas para ambas regiones se observaron para el ácido palmitoleico y los índices C18:0/C16:0 y C18:1(n-7)/C16:1(n-7). Los genes MAML3 y SETD7 fueron estudiados como genes candidatos posicionales para la región localizada a 93 Mb. Los dos nuevos microsatélites analizados en el gen MAML3 y el SNP del gen SETD7 (SETD7:c.700G>T) no mostraron las asociaciones más significativas en esta región, descartando estos polimorfismos como las mutaciones causales. Además, en la región localizada a 119 Mb, el SNP ELOVL6:c.-533C>T mostró la asociación más significativa con el porcentaje de los ácidos palmítico y palmitoleico y los índices de elongación en GD. Los resultados obtenidos para el gen ELOVL6, gen candidato posicional del QTL localizado a 119 Mb refuerzan la hipótesis de su efecto pleiotrópico sobre la composición de AG en GD y en músculo, y su papel en la determinación del QTL del SSC8 para el perfil de AG. Por otra parte, se utilizaron datos del genoma completo de cerdos ibéricos y landrace para identificar 1.279 variaciones en el número de copias (CNV), las cuales se fusionaron en 540 regiones de CNVs (CNVRs). El impacto de cuatro de ellas fue estudiado para caracteres de crecimiento y composición de AG. Se encontró asociación con la longitud de la canal y la composición de AG en grasa intramuscular y GD para el CNVR112. Este CNVR contiene el gen GPAT2 que cataliza la biosíntesis de triglicéridos y glicerofosfolípidos. Los resultados obtenidos subrayan la importancia de los CNVRs en la determinación de caracteres económicamente importantes en el cerdo. Finalmente, se analizó la expresión de 44 genes candidatos relacionados con el metabolismo lipídico en 115 animales. El estudio de asociación genómico con los datos de expresión (eGWAS) identificó 193 eSNPs localizados en 19 eQTLs. Tres de los eQTLs correspondientes a los genes ACSM5, FABP4 y FADS2 se clasificaron como cis-eQTLs; mientras que los 16 eQTLs restantes mostraron efectos reguladores en trans. Estos hallazgos, junto con los polimorfismos evaluados para alguno de estos genes, mejoran nuestro conocimiento sobre los mecanismos funcionales implicados en la variación de los caracteres relacionados con la calidad de la carne porcina., Pork is one of the main sources of human-consumed meat and consumer’s preference towards high quality meat is increasing. Hence, understanding the molecular mechanisms affecting meat production and quality would help in the selection of these traits. Meat quality is determined largely by its fatty acid (FA) composition and understanding the underlying molecular processes of FA composition is the general objective of this thesis. We analyzed quantitative trait loci (QTL) on porcine chromosome 8 (SCC8) for FA composition in backfat, identifying two trait-associated SNP regions at 93 Mb and 119 Mb. The strongest statistical signals for both regions were observed for palmitoleic acid and, C18:0/C16:0 and C18:1(n-7)/C16:1(n-7) elongation ratios. MAML3 and SETD7 genes were analyzed as positional candidate genes in the 93 Mb region. The two novel microsatellites analyzed in the MAML3 gene, and the SETD7:c.700G>T SNP in the SETD7 gene did not show the strongest signal in this region, discarding these polymorphisms as the causal mutations. Furthermore, in the 119 Mb region, the ELOVL6:c.-533C>T SNP showed a strong association with the percentage of palmitic and palmitoleic acids and elongation ratios in backfat. These results for ELOVL6 gene, support the hypothesis that it has a pleiotropic effect in backfat and muscle for the 119 Mb QTL, and reinforce this gene as a strong candidate for the SSC8 QTL for FA composition. Moreover, whole genome sequence (WGS) data from Iberian and Landrace pigs were used to identify 1,279 copy number variations (CNVs), merging into 540 swine CNV regions (CNVRs). The impact of four of them in growth and FA composition in intramuscular fat and backfat was studied. Association with carcass length and FA composition in backfat and intramuscular fat was showed for the CNVR112, containing the GPAT2 gene which catalyse the biosynthesis of triglycerides and glycerophospholipids. These results underline the importance of CNVRs affecting economically important traits in pigs. Finally, the adipose tissue mRNA expression of 44 candidate genes related with lipid metabolism was analyzed in 115 animals. The expression genome-wide association (eGWAS) identified 193 eSNPs located in 19 expression QTLs (eQTLs). Three out of 19 eQTLs corresponding to ACSM5, FABP4, and FADS2 were classified as cis-acting eQTLs, whereas the remaining 16 eQTLs had trans-regulatory effects. These findings and the polymorphisms evaluated for some of these genes provide new data to further understand the functional mechanisms implicated in the variation of meat quality traits in pigs.

La carne de cerdo es la más consumida mundialmente. Los programas de mejora animal se han centrado en realizar una producción sostenible y eficiente mejorando tanto los caracteres de reproducción, como de crecimiento y calidad de la carne. Actualmente no se dispone de un conocimiento completo sobre la base genética que determina estos caracteres de producción y por este motivo el objetivo principal de esta tesis consistió en profundizar en los mecanismos moleculares que determinan los caracteres de crecimiento y calidad de la carne. Los genes FABP4 y FABP5 fueron evaluados como genes candidatos para la composición de ácidos grasos en músculo y grasa dorsal. Los animales portadores del alelo C para el polimorfismo FABP4: g.2634_2635insC mostraron un mayor contenido intramuscular de ácidos grasos C16:0 y C16:1(n-7) y, un menor contenido de C18:2(n-6) y una mayor expresión del gen FABP4 en grasa dorsal. Dicho polimorfismo se encuentra dentro de una diana de unión para el gen PPARG pudiendo este factor nuclear determinar las diferencias de expresión observadas. Respecto la expresión del gen FABP5 en músculo, se detectaron regiones asociadas en los SSC4, SSC6, SSC9 y SSC13. Sin embargo, no se observó una clara asociación de genotipos del SNP FABP5: g.3000T> G con la composición de ácidos grasos. Con el objetivo de explorar el transcriptoma del músculo para identificar genes y rutas metabólicas relacionadas con el contenido y la composición de ácidos grasos en músculo, se secuenció el ARN de dos grupos de animales que diferían en estos caracteres. Se identificó un total de 131 genes diferencialmente expresados entre grupos en su mayoría relacionados con las rutas del metabolismo lipídico. El análisis funcional mostró una menor captación de glucosa y una inhibición de la lipogénesis en los animales con un mayor contenido en PUFA. Los enfoques utilizados hasta ahora para el análisis de los caracteres complejos implican generalmente varios genes con efectos pequeños y no consideran las interacciones funcionales. Por esta razón, se decidió aplicar un enfoque de redes génicas basada en el análisis de co-asociación entre SNPs para el estudio de caracteres relacionados con el crecimiento, la conformación y el engrasamiento. Se identificaron tres factores de transcripción, PRDM16, ELF1 y PPARG, como principales reguladores de la red. Además, 54 de los genes identificados en la red pertenecían a ontologías relacionadas con el crecimiento. Finalmente, con el objetivo de evaluar genes candidatos funcionales identificados en estudios previos de nuestro grupo afectando a caracteres de deposición y composición de ácidos grasos en músculo, se analizó la expresión de 45 genes en 114 animales. El eGWAS identificó 241 eSNPs distribuidos en 18 eQTLs. Tres de los 18 eQTLs presentaban variantes en cis, y en 16 de los eQTLs se identificaron zonas reguladoras en trans. Los resultados permitieron identificar potenciales reguladores y mejorar nuestro conocimiento sobre los mecanismos funcionales implicados en estos caracteres complejos., Pork is the most consumed meat worldwide. The breeding programs have been working towards a sustainable pig production improving reproduction traits, growth and meat quality traits. The genetic basis determining these complex and economically important production traits remains elusive not being fully understood. In the present thesis we pursue to clarify the molecular mechanisms that determine growth and meat quality traits. The functional role of FABP4 and FABP5 genes in determining the fatty acid composition in muscle and backfat tissues was evaluated. Animals inheriting the C allele for FABP4:g.2634_2635insC polymorphism showed higher intramuscular content of C16:0 and C16:1(n-7) fatty acids and, decreased content of C18:2(n-6) fatty acid and higher FABP4 expression in backfat. Moreover, FABP4:g.2634_2635insC was located inside a PPARG binding site suggesting a role of this nuclear receptor in the regulation of FABP4 gene expression. The FABP5:g.3000T>G SNP was the most significant marker for intramuscular percentages of C18:1(n-9), C18:2(n-6), and MUFA. However, this variant was not associated with FABP5 gene expression, being FABP5 gene expression in muscle regulated by other genomic regions located on SSC4, SSC6, SSC9 and SSC13. Aiming to identify genes and pathways affecting the intramuscular fatty acid composition, the muscle transcriptome of two groups of pigs with extreme phenotypes for these traits was analyzed. A total of 131 genes mostly related to lipid metabolism pathways were identified as differentially expressed between groups. The functional analysis showed that animals with a higher content of PUFA presented low fatty acid and glucose uptake resulting in an inhibition of the lipogenesis. Gene-by-gene approaches used until now are limited when analyzing complex traits usually implying several genes with small effects; moreover, they ignore functional interactions. For this reason, we decided to apply a gene network approach based on SNP-by-SNP co-association analysis to explore growth, conformation and fatness related traits. From the resulting network formed by 513 nodes and 639 edges, three transcription factors PRDM16, ELF1 and PPARG were likely to be the major regulators of this network. Moreover, 54 genes identified within the network belonged to growth-related ontologies. Finally, with the aim to evaluate functional candidate genes affecting intramuscular deposition and fatty acid composition traits from previous studies of our group, the mRNA expression of 45 genes was measured in 114 animals. The eGWAS identified 241 eSNPs distributed in 18 eQTLs. Three out of 18 eQTLs presented cis-acting variants and 16 eQTLs showed trans regulatory effects. The results highlighted putative key regulators and improved our knowledge in the functional regulatory mechanisms implicated in these complex traits.