Fat Content In Pigs Research Articles

Identification of a Candidate Gene Regulating Intramuscular Fat Content in Pigs through the Integrative Analysis of Transcriptomics and Proteomics Data.

Pork is a widely consumed source of animal protein worldwide, and the intramuscular fat (IMF) content in pork plays a crucial role in determining its quality. In this study, we sought to identify candidate genes that regulate IMF deposition in pigs. We performed tandem mass tags (TMT)-based quantitative proteomics analysis using Longissimus dorsi (LD) muscle samples obtained from eight pigs with extremely high and low IMF content among a group of 28 Duroc pigs and identified 50 differentially abundant proteins (DAPs). Additionally, we compared the proteomics data with RNA-sequencing data obtained in our previous study and identified TUSC5 as a differentially expressed gene corresponding to the relevant DAP. To investigate the potential role of TUSC5 in adipogenesis, we suppressed TUSC5 expression in mouse 3T3-L1 preadipocytes using short hairpin RNA (shRNA) and observed a significant reduction in the differentiation of 3T3-L1 cells into adipocytes, as indicated by Oil Red O staining and triglyceride content. Moreover, we observed a reduction in the expression of genes associated with adipogenesis (PPARG, CEBPA, FABP4, and FASN) following TUSC5 suppression. Through an integrative analysis of transcriptomics and proteomics data, our study identified TUSC5 as a crucial candidate gene associated with the regulation of IMF content in pigs.

Dietary calcium supplementation promotes the accumulation of intramuscular fat

BackgroundIn the livestock industry, intramuscular fat content is a key factor affecting meat quality. Many studies have shown that dietary calcium supplementation is closely related to lipid metabolism. However, few studies have examined the relationship between dietary calcium supplementation and intramuscular fat accumulation.MethodsHere, we used C2C12 cells, C57BL/6 mice (n = 8) and three-way cross-breeding pigs (Duroc×Landrace×Large white) (n = 10) to study the effect of calcium addition on intramuscular fat accumulation. In vitro, we used calcium chloride to adjust the calcium levels in the medium (2 mmol/L or 3 mmol/L). Then we measured various indicators. In vivo, calcium carbonate was used to regulate calcium levels in feeds (Mice: 0.5% calcium or 1.2% calcium) (Pigs: 0.9% calcium or 1.5% calcium). Then we tested the mice gastrocnemius muscle triglyceride content, pig longissimus dorsi muscle meat quality and lipidomics.ResultsIn vitro, calcium addition (3 mmol/L) had no significant effect on cell proliferation, but promoted the differentiation of C2C12 cells into slow-twitch fibers. Calcium supplementation increased triglyceride accumulation in C2C12 cells. Calcium addition increased the number of mitochondria and also increased the calcium level in the mitochondria and reduced the of key enzymes activity involved in β-oxidation such as acyl-coenzyme A dehydrogenase. Decreasing mitochondrial calcium level can alleviate lipid accumulation induced by calcium addition. In addition, calcium addition also reduced the glycolytic capacity and glycolytic conversion rate of C2C12 cells. In vivo, dietary calcium supplementation (1.2%) promoted the accumulation of triglycerides in the gastrocnemius muscle of mice. Dietary calcium supplementation (1.5%) had no effect on pig weight, but significantly improved the flesh color of the longissimus dorsi muscle, reduced the backfat thickness and increased intramuscular fat content in pigs. Besides, calcium addition had no effect on longissimus dorsi pH, electrical conductivity and shear force.ConclusionsThese results suggest that calcium addition promotes intramuscular fat accumulation by inhibiting the oxidation of fatty acids. These findings provide a new tool for increasing intramuscular fat content and an economical strategy for improving meat quality.

Evidence Against the Causal Relationship Between a Putative Cis-Regulatory Variant of MYH3 and Intramuscular Fat Content in Pigs.

Improving meat quality has become the main goal of modern pig breeding. Intramuscular fat content (IMF) is an important trait influencing meat quality of livestock, but the molecular mechanism behind this trait is still unclear. Recently, Cho et al. reported the discovery of the first causal mutation affecting IMF and red flesh color (a*) in pigs, namely XM_013981330.2:g.−1805_−1810del, a 6-bp deletion variant in the porcine MYH3 promoter region. The objective of this study was to reassess the causality of this mutation for its potential commercial application. By Sanger sequencing, we firstly identified several new variants (including a 4-bp deletion) at or near the 6-bp deletion site, which formed four haplotypes in multiple breeds. Unexpectedly, the 6-bp deletion allele, previously determined as the MYH3 Q allele because of its significantly positive effect on IMF and a*, was found not only in Chinese indigenous breeds, but also in four western commercial breeds with relatively lower IMF levels, including Duroc, Large White, Landrace and Pietrain. More surprisingly, we found that the MYH3 Q allele and the haplotypes harboring it had no significant effects on IMF, marbling and color score in three large-scale divergent pig populations: the heterogeneous F6 and F7 pigs and commercial crossbred Duroc × (Landrace × Yorkshire) pigs. Transient transfection analysis in porcine satellite cells showed that the 6-bp deletion variants had a negligible effect on transcription of reporter gene, but could attenuate the MRF (myogenesis regulatory factors)-induced increase in luciferase activity of the MYH3 promoter vector. The MYH3 protein level in muscle did not differ significantly among the haplotype groups. Therefore, our results cannot support the causal relationship between the 6-bp deletion in MYH3 and IMF trait, suggesting that the causal mutation for the IMF QTL on SSC12 needs to be further identified.

Integrated Microbiome and Metabolome Analysis Reveals a Positive Change in the Intestinal Environment of Myostatin Edited Large White Pigs.

Myostatin (MSTN) functional inactivation can change the proportion of lean meat and fat content in pigs. While both genotype and microbial composition are known to affect the host phenotype, so far there has been no systematic study to detect the changes in the intestinal microbial composition and metabolome of MSTN single copy mutant pigs. Here, we used 16S rDNA sequencing and metabolome analysis to investigate how MSTN gene editing affects changes in the microbial and metabolome composition in the jejunum and the cecum of Large White pigs. Our results showed that Clostridium_sensu_stricto_1, Bifidobacterium, Lachnospiraceae_UCG-007, Clostridium_sensu_stricto_6, Ruminococcaceae_UCG-002, and Ruminococcaceae_UCG-004 were significantly upregulated; while Treponema_2 and T34_unclassified were significantly downregulated in the jejunum of MSTN pigs. Similarly, Phascolarctobacterium, Ruminiclostridium_9, Succinivibrio, Longibaculum, and Candidatus_Stoquefichus were significantly upregulated, while Barnesiella was significantly downregulated in the cecum of MSTN pigs. Moreover, metabolomics analysis showed significant changes in metabolites involved in purine, sphingolipid and tryptophan metabolism in the jejunum, while those associated with glycerophospholipid and pyrimidine metabolism were changed in the cecum. Spearman correlation analysis further demonstrated that there was a significant correlation between microflora composition and metabolites. Our analyses indicated the MSTN editing affects the composition of metabolites and microbial strains in the jejunum and the cecum, which might provide more useable nutrients for the host of MSTN± Large White pigs.

Investigation on the Effect of Two Fat Metabolism Related Pathways on Intramuscular Fat Content in Pigs

Investigation on the Effect of Two Fat Metabolism Related Pathways on Intramuscular Fat Content in Pigs

Promoter CpG methylation status affects ADRP gene expression level and intramuscular fat content in pigs

Adipose differentiation-related protein (ADRP) is a lipid droplet-associated protein that is expressed early during adipose differentiation. However, the underlying mechanism is still unclear. In this study, the porcine ADRP (pADRP) gene regulatory mechanism and its relationship to intramuscular fat (IMF) content are analysed. We investigated the characteristics of the pADRP gene prompter as well as its transcription level, core promoter and epigenetic modification status using real-time PCR, Dual-luciferase reporter assays and bisulphite sequencing. The results showed that pADRP mRNA expression in Longissimus Dorsi (LD) muscle of the low IMF (LEAN) group was lower than that in the high IMF (FAT) group (p<.01). Maximum promoter activity was found in the −926 bp to −235 bp region. The methylation level of the pADRP of this region (mainly 7th, 10th, 12th CpG sites) in the low IMF group (86.67%) was significantly higher than that in the high IMF group (46.7%) (p<.01). Our result plays an important role in pADRP transcriptional regulation and provides a good prospect for the marbling traits improvement. Highlights pADRP expression differs in divergent IMF content pigs. Promoter region located −929 to −235 bp, and its methylation contributes to gene expression.

TMT-labeled quantitative proteomic analyses on the longissimus dorsi to identify the proteins underlying intramuscular fat content in pigs

The Laiwu pig is famous for its excessively extremely high level of intramuscular fat content (IMF), however, the exact regulatory mechanism underlying intramuscular fat deposition in skeletal muscle is still unknown. As an economically important trait in pigs, IMF is controlled by multiple genes and biological pathways. In this study, we performed an integrated transcriptome-assisted TMT-labeled quantitative proteomic analysis of the longissimus dorsi (LD) muscle in Laiwu pigs at the fastest IMF deposition stage and identified 5074 unique proteins and 52 differentially abundant proteins (DAPs) (>1.5-fold cutoff, p < .05). These DAPs were hierarchically clustered in the LD muscle over two developmental stages from 120 d to 240 d. A comparison between transcriptomic (mRNA) and proteomic data revealed two differentially expressed genes corresponding to the DAPs. Changes in the levels of the nine proteins were further analyzed using RT-qPCR and parallel reaction monitoring (PRM). The proteins identified in this study could serve as candidates for elucidating the molecular mechanism of IMF deposition in pigs. SignificanceThe intramuscular fat content (IMF) refers to the amount of fat within muscles and plays an important role in meat quality by affecting meat quality-related traits, such as tenderness, juiciness and flavor. Using the integrated transcriptome-assisted TMT-labeled quantitative proteomic approach to characterize changes in the proteomic profile of the longissimus dorsi muscle, we identified differentially abundant proteins, such as ALDH1B1, OTX2, AnxA6 and Zfp512, that are associated with intramuscular fat deposition and fat biosynthesis in pigs. These proteins could serve as candidates for elucidating the molecular mechanism of IMF deposition in pigs.

A functional regulatory variant of MYH3 influences muscle fiber-type composition and intramuscular fat content in pigs.

Muscle development and lipid accumulation in muscle critically affect meat quality of livestock. However, the genetic factors underlying myofiber-type specification and intramuscular fat (IMF) accumulation remain to be elucidated. Using two independent intercrosses between Western commercial breeds and Korean native pigs (KNPs) and a joint linkage-linkage disequilibrium analysis, we identified a 488.1-kb region on porcine chromosome 12 that affects both reddish meat color (a*) and IMF. In this critical region, only the MYH3 gene, encoding myosin heavy chain 3, was found to be preferentially overexpressed in the skeletal muscle of KNPs. Subsequently, MYH3-transgenic mice demonstrated that this gene controls both myofiber-type specification and adipogenesis in skeletal muscle. We discovered a structural variant in the promotor/regulatory region of MYH3 for which Q allele carriers exhibited significantly higher values of a* and IMF than q allele carriers. Furthermore, chromatin immunoprecipitation and cotransfection assays showed that the structural variant in the 5′-flanking region of MYH3 abrogated the binding of the myogenic regulatory factors (MYF5, MYOD, MYOG, and MRF4). The allele distribution of MYH3 among pig populations worldwide indicated that the MYH3 Q allele is of Asian origin and likely predates domestication. In conclusion, we identified a functional regulatory sequence variant in porcine MYH3 that provides novel insights into the genetic basis of the regulation of myofiber type ratios and associated changes in IMF in pigs. The MYH3 variant can play an important role in improving pork quality in current breeding programs.

Expression of NR3C1, INSR and SLC2A4 genes in skeletal muscles and CBG in liver depends on age and breed of pigs

The genes encoding glucose transporter 4 (SLC2A4, GLUT4), insulin receptor (INSR) and glucocorticoid receptor (NR3C1) are considered regulators of energy metabolism that may influence fat content in skeletal muscle at different ages and breeds of pigs. In the study we performed analysis of expression of NR3C1, INSR and SLC2A4 genes in two skeletal muscle tissues: semimembranosus and longissimus dorsi muscle in gilts from three breeds of pigs that differed in intramuscular fat content: Duroc (DU), Pulawska (PUL) and Polish Large White (PLW) at 60, 90, 120, 150, 180 and 210 days after weaning. We also analyzed the expression of cortisol binding globulin (CBG) in the liver. Expression was analyzed with real time PCR (qPCR) using a relative quantification method. In blood, the concentration of cortisol, insulin and leptin were evaluated with radioimmunoassay. The concentration of metabolites (triglycerides and glucose) related to the changes of lipids content was also measured. We observed a significant relationship between the breed and the age of pigs and the expression of analyzed genes in muscle tissues (P &amp;lt; 0.01 for all analyzed genes) as well as with two hormones (for insulin P = 0.001; for cortisol P &amp;lt; 0.0001). Leptin level and metabolites in serum were not significant in the model (P &amp;gt; 0.05). The expression of analyzed genes (SLC2A4, INSR and NR3C1 in muscles and CBG in liver) strongly correlates with intramuscular fat content in pigs and is associated with age and breed.

Linoleic acid metabolic pathway allows for an efficient increase of intramuscular fat content in pigs

BackgroundIntramuscular fat (IMF) content is a relevant trait for high-quality meat products such as dry-cured ham, but increasing IMF has the undesirable correlated effect of decreasing lean growth. Thus, there is a need to find selection criteria for IMF independent from lean growth. In pigs, the proportion of linoleic (C18:2) and arachidonic (C20:4) acids decline with fat deposition and therefore they can be considered as indicators of fatness. The aim of this research was to estimate the genetic variation for C18:2 and C20:4 in IMF and their genetic correlations with IMF and lean growth traits, with the objective to assess their potential as specific biomarkers of IMF. The analysis was conducted using a full-pedigreed Duroc resource line with 91,448 records of body weight and backfat thickness (BT) at 180 days of age and 1371 records of fatty acid composition in the muscle gluteus medius.ResultsThe heritability estimates for C18:2 and C20:4 in IMF, whether expressed in absolute (mg/g of muscle) or in relative (mg/g of fatty acid) terms, as well as for their ratio (C20:4/C18:2), were high (> 0.40), revealing that the C18:2 to C20:4 pathway is subjected to substantial genetic influence. Litter effects were not negligible, with values ranging from 8% to 15% of the phenotypic variance. The genetic correlations of C18:2 and C20:4 with IMF and BT were negative (− 0.75 to − 0.66, for IMF, and − 0.64 to − 0.36, for BT), if expressed in relative values, but almost null (− 0.04 to 0.07), if expressed in absolute values, except for C18:2 with IMF, which was highly positive (0.88). The ratio of C20:4 to C18:2 also displayed a stronger genetic correlation with IMF (− 0.59) than with BT (− 0.10).ConclusionsThe amount of C18:2 in muscle can be used as an IMF-specific biomarker. Selection for the absolute amount of C18:2 is expected to deliver a similar response outcome as selection for IMF at restrained BT. Further genetic analysis of the C18:2 metabolic pathway may provide new insights into differential fat deposition among adipose tissues and on candidate genes for molecular markers targeting specifically for one of them.

Data Mining and Validation of AMPK Pathway as a Novel Candidate Role Affecting Intramuscular Fat Content in Pigs.

Simple SummaryIntramuscular fat (IMF) is increasingly being recognized as a key meat trait in the modern pork industry. The aims of this research were to identify potential signaling pathways associated with IMF content in the longissimus dorsi (LD) muscle of different pig breeds and investigate the gene expression levels in the screened signaling pathways. Our results indicated that the AMPK signaling pathway may be related to IMF deposition in the LD muscle of pigs. The results of qRT-PCR analysis showed that the expression of ten key hub genes (AMPK, ADIPOR1, ADIPOR2, LKB1, CAMKKβ, CPT1A, CPT1B, PGC-1α, CD36, and ACC1) differed between the LD muscle of Min and Large White pigs. The protein expression levels of AMPK, LKB1, CaMKK2, CPT1A, and ACC1 were similar to the genes expression patterns in the LD muscle of Large White pigs. The results of this study provide novel insights into the regulatory function of the AMPK signaling pathway in relation to the IMF content in the LD muscle of different pigs.Intramuscular fat (IMF) is an important economic trait for pork quality and a complex quantitative trait regulated by multiple genes. The objective of this work was to investigate the novel transcriptional effects of a multigene pathway on IMF deposition in the longissimus dorsi (LD) muscles of pigs. Potential signaling pathways were screened by mining data from three gene expression profiles in the Gene Expression Omnibus (GEO) database. We designed quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) arrays for the candidate signaling pathways to verify the results in the LD muscles of two pig breeds with different IMF contents (Large White and Min). Western blot analysis was used to detect the expression levels of several candidate proteins. Our results showed that the AMPK signaling pathway was screened via bioinformatics analysis. Ten key hub genes of this signaling pathway (AMPK, ADIPOR1, ADIPOR2, LKB1, CAMKKβ, CPT1A, CPT1B, PGC-1α, CD36, and ACC1) were differentially expressed between the Large White and Min pigs. Western blot analysis further confirmed that LKB1/CaMKK2-AMPK-ACC1-CPT1A axis dominates the activity of AMPK signaling pathway. Statistical analyses revealed that AMPK signaling pathway activity clearly varied among the two pig breeds. Based on these results, we concluded that the activation of the AMPK signaling pathway plays a positive role in reducing IMF deposition in pigs.

Identification and Functional Analysis of Long Intergenic Non-coding RNAs Underlying Intramuscular Fat Content in Pigs.

Intramuscular fat (IMF) content is an important trait that can affect pork quality. Previous studies have identified many genes that can regulate IMF. Long intergenic non-coding RNAs (lincRNAs) are emerging as key regulators in various biological processes. However, lincRNAs related to IMF in pig are largely unknown, and the mechanisms by which they regulate IMF are yet to be elucidated. Here we reconstructed 105,687 transcripts and identified 1,032 lincRNAs in pig longissimus dorsi muscle (LDM) of four stages with different IMF contents based on published RNA-seq. These lincRNAs show typical characteristics such as shorter length and lower expression compared with protein-coding genes. Combined with methylation data, we found that both the promoter and genebody methylation of lincRNAs can negatively regulate lincRNA expression. We found that lincRNAs exhibit high correlation with their protein-coding neighbors in expression. Co-expression network analysis resulted in eight stage-specific modules, gene ontology and pathway analysis of them suggested that some lincRNAs were involved in IMF-related processes, such as fatty acid metabolism and peroxisome proliferator-activated receptor signaling pathway. Furthermore, we identified hub lincRNAs and found six of them may play important roles in IMF development. This work detailed some lincRNAs which may affect of IMF development in pig, and facilitated future research on these lincRNAs and molecular assisted breeding for pig.

Molecular characterization, tissue expression profile, and SNP analysis of porcine SLC13A5.

Solute carrier family 13 (sodium-dependent citrate transporter member 5, SLC13A5) gene has been recently found to play an important role in intramuscular fat content in pigs. In this study, the full-length cDNA of porcine SLC13A5 was obtained from the longissimus dorsi muscle of Shaziling pigs, using the rapid amplification of cDNA ends technique. Full-length porcine SLC13A5 cDNA was 2118 bp, with a 1665-bp open reading frame encoding 554 amino acids. The porcine SLC13A5 protein was analyzed using bioinformatic methodology, and found to include 18 potential phosphorylation sites (including six serine, nine threonine, and three tyrosine) and eight putative transmembrane domains. One single nucleotide polymorphism (SNP) site, A251G, was identified by polymerase chain reaction (PCR)-restriction fragment length polymorphism and the associations of this SNP with age at 100 kg and corrected back fat thickness were found to be not significant. Expression of SLC13A5 was evaluated in ten tissues from 25-day-old full-sib Yorkshire and Shaziling piglets (both N = 3), using quantitative PCR analysis. Expression levels of SLC13A5 differed significantly between the breeds in cecum, liver and crureus muscle. In each breed, gene expression levels were significantly different in longissimus dorsi muscle, compared to the nine other tissues. This study has laid the foundation for further investigations of the molecular mechanisms of SLC13A5 in pigs.

Neuronal Genes for Subcutaneous Fat Thickness in Human and Pig Are Identified by Local Genomic Sequencing and Combined SNP Association Study

Obesity represents a major global public health problem that increases the risk for cardiovascular or metabolic disease. The pigs represent an exceptional biomedical model related to energy metabolism and obesity in humans. To pinpoint causal genetic factors for a common form of obesity, we conducted local genomic de novo sequencing, 18.2 Mb, of a porcine QTL region affecting fatness traits, and carried out SNP association studies for backfat thickness and intramuscular fat content in pigs. In order to relate the association studies in pigs to human obesity, we performed a targeted genome wide association study for subcutaneous fat thickness in a cohort population of 8,842 Korean individuals. These combined association studies in human and pig revealed a significant SNP located in a gene family with sequence similarity 73, member A (FAM73A) associated with subscapular skin-fold thickness in humans (rs4121165, GC-corrected p-value = 0.0000175) and with backfat thickness in pigs (ASGA0029495, p-value = 0.000031). Our combined association studies also suggest that eight neuronal genes are responsible for subcutaneous fat thickness: NEGR1, SLC44A5, PDE4B, LPHN2, ELTD1, ST6GALNAC3, ST6GALNAC5, and TTLL7. These results provide strong support for a major involvement of the CNS in the genetic predisposition to a common form of obesity.

Candidate gene expression and intramuscular fat content in pigs

Seventy-two pigs of three genetic groups (Brazilian indigenous breed Piau, Commercial line and Crossbred) of both sexes were slaughtered at four live weights (30, 60, 90 and 120 kg). Intramuscular fat (IMF) content in Longissimus dorsi muscle of each animal was extracted and correlated with candidate gene mRNA expression (ATN1, EEF1A2, FABP3, LDLR, MGP, OBSCN, PDHB, TRDN and RYR1). Within slaughter weight of 120 kg, Piau and Crossbred pigs showed higher IMF content (p < 0.05) than commercial animals, with 2.48, 2.08 and 1.00% respectively. Barrows presented higher values of IMF (p < 0.05) than gilts (1.54 and 1.30% respectively). Gene expression of EEF1A2, FABP3, LDLR, OBSCN, PDHB, TRDN and RYR1 were correlated with IMF (p < 0.05) using the whole dataset. For Piau data only, expression of FABP3, LDLR, MGP, OBSCN, PDHB, TRDN and RYR1 showed correlation with IMF (p < 0.05). Genes that have important roles in lipid transportation inside the cell (FABP3) and tissues (LDLR) showed correlation with IMF of, respectively, 0.68 and 0.63 using the whole data set, and 0.90 and 0.91 using data from Piau animals. The highly positive correlation of the LDLR and FAPB3 expression with IMF content may confirm that these genes are important for fat deposition in the porcine L. dorsi muscle.

Muscle transcriptomic profiles in pigs with divergent phenotypes for fatness traits

BackgroundSelection for increasing intramuscular fat content would definitively improve the palatability and juiciness of pig meat as well as the sensorial and organoleptic properties of cured products. However, evidences obtained in human and model organisms suggest that high levels of intramuscular fat might alter muscle lipid and carbohydrate metabolism. We have analysed this issue by determining the transcriptomic profiles of Duroc pigs with divergent phenotypes for 13 fatness traits. The strong aptitude of Duroc pigs to have high levels of intramuscular fat makes them a valuable model to analyse the mechanisms that regulate muscle lipid metabolism, an issue with evident implications in the elucidation of the genetic basis of human metabolic diseases such as obesity and insulin resistance.ResultsMuscle gene expression profiles of 68 Duroc pigs belonging to two groups (HIGH and LOW) with extreme phenotypes for lipid deposition and composition traits have been analysed. Microarray and quantitative PCR analysis showed that genes related to fatty acid uptake, lipogenesis and triacylglycerol synthesis were upregulated in the muscle tissue of HIGH pigs, which are fatter and have higher amounts of intramuscular fat than their LOW counterparts. Paradoxically, lipolytic genes also showed increased mRNA levels in the HIGH group suggesting the existence of a cycle where triacylglycerols are continuously synthesized and degraded. Several genes related to the insulin-signalling pathway, that is usually impaired in obese humans, were also upregulated. Finally, genes related to antigen-processing and presentation were downregulated in the HIGH group.ConclusionOur data suggest that selection for increasing intramuscular fat content in pigs would lead to a shift but not a disruption of the metabolic homeostasis of muscle cells. Future studies on the post-translational changes affecting protein activity or expression as well as information about protein location within the cell would be needed to to elucidate the effects of lipid deposition on muscle metabolism in pigs.

Acetyl-CoA carboxylase and stearoyl-CoA desaturase protein expression in subcutaneous adipose tissue is reduced in pigs selected for decreased backfat thickness at constant intramuscular fat content.

The objectives of this study were 1) to determine whether selection toward less subcutaneous fat thickness at constant intramuscular fat content in pigs is related to tissue-specific changes in the expression of lipogenic enzymes acetyl-CoA carboxylase (ACC), stearoyl-CoA desaturase (SCD), and Delta(6)-desaturase (Delta6d); and 2) to investigate tissue specific distribution of the porcine ACC, SCD, and Delta6d. The study was conducted on 20 purebred Duroc barrows. Ten animals were from a group selected for decreased subcutaneous fat thickness at constant intramuscular fat content (experimental group). The other 10 animals were from the unselected (control) group. Distribution of ACC, SCD, and Delta6d was investigated in semimembranosus muscle (SM), subcutaneous adipose tissue (SA), liver (L), kidney (K), heart (H), diaphragm (D), rectus capitis muscle (RCM), and abdominal fat (AF). The enzyme expression was studied in 10 animals in the case of SM and SA and in 4 animals in the case of other tissues. The following expression pattern was established for ACC: SM <or= H = K <or= D < RCM < L < AF = SA, whereas the expression patterns for SCD and Delta6d proteins were SM < H < RCM < D < L < K < AF = SA and RCM = SM = D < L <or= H < SA < K < AF, respectively. Expression of ACC and SCD proteins was less in subcutaneous adipose tissue of the experimental animals when compared with the control group (P < 0.001). However, no difference (P > 0.1) in ACC and SCD protein expression between the control and experimental groups was observed in SM. Expression of Delta6d protein did not differ between the control and experimental groups for SA (P = 0.47) or SM (P = 0.31). There was a positive relationship between muscle SCD protein expression and intramuscular fat content (r = 0.48, P < 0.05). Intramuscular fat content did not correlate with ACC or Delta6d protein expression (P = 0.23 and P = 0.80, respectively). We conclude that SCD might be an effective potential biomarker for intramuscular fat deposition.

An age-dependent association between a leptin C3469T single nucleotide polymorphism and intramuscular fat content in pigs

The objective of this work was to investigate whether the C3469T mutation at exon 3 of the pig leptin ( Lep) gene is associated with fatness and if the magnitude and direction of these associations is consistent within age. Repeated measurements on body weight (BW), ultrasonic subcutaneous backfat (SB) and loin-muscle (LT) thickness, and intramuscular fat (IMF) content and composition were taken at 160, 180, 215 and 225 days of age on 162 Duroc barrows previously genotyped for the C3469T mutation. Plasma leptin concentration was determined at 180 days. A total of 505 records of IMF content and composition, which were determined both in live samples of longisimus dorsi muscle (LM) and in carcass samples of LM and gluteus medius muscle, were used in the analyses. The genotypic frequencies associated with the C3469T Lep mutation were 0.76, 0.23, and 0.01 for TT, TC, and CC genotypes, respectively. The effect of the Lep genotypes was assessed using a Bayesian approach based on a polynomial mixed model on age. No evidence was found for an association of the Lep genotypes neither with BW, SB thickness nor with plasma leptin concentration. The posterior mean of the difference TT–TC ranged, for LT, from 0.82 (160 days, posterior probability density (PP)> 0 of 87%) to 1.98 mm (220 days, PP > 0 of 96%) and, for IMF, from 0.96 (160 days, PP > 0 of 84%) to − 1.64 mg/g DM (220 days, PP > 0 of 2%). The results suggest an age-dependent association of the C3469T Lep polymorphism with LT and IMF, which may be useful to explain some controversial results in the literature concerning the association of this polymorphism with IMF content.

Heart fatty acid binding protein is upregulated during porcine adipocyte development1

Heart fatty acid binding protein (H-FABP) has been associated with intramuscular fat content in pigs. In the current study, we showed that expression of H-FABP mRNA in adipose tissue of adult pigs was 8.5% of that in heart and 30% of that in skeletal muscle, and that H-FABP mRNA level was more than 10% of that of adipocyte fatty acid binding protein mRNA in adipose tissue. Levels of H-FABP mRNA reached a maximum in adipose tissue from 7-d neonates, with no further increase in the adult. Also, H-FABP mRNA was induced during adipogenic differentiation of stromal-vascular cells derived from adipose tissue and skeletal muscle. In conclusion, H-FABP may play a role in adipose tissue development and function in the pig.

Liver X receptor alpha and beta genes have the potential role on loin lean and fat content in pigs

Liver X receptor alpha (LXRA) and beta (LXRB) are members of the nuclear receptor subfamily and are important regulators of genes involved in lipid, fatty acid and glucose metabolism in liver, and adipose tissue as well as in skeletal muscle. To investigate whether the two LXR genes play a role in influencing lean and fat growth in pigs, we discovered and examined two polymorphisms in LXRA (LXRA Bsl in exon 2, and LXRA HpyCH4 III in intron 8) and one polymorphism in LXRB (LXRB Aci I in exon 5) for genetic linkage and association analyses. Linkage analyses using a three-generation resource family of a cross between the Berkshire and Yorkshire (BY) pig breeds assigned LXRA to SSC2 and LXRB to SSC6. Association analyses were carried out among those polymorphisms and traits evaluated in the BY F(2) family and four pig commercial populations. These analyses indicated that the LXRA HpyCH4 III polymorphism was significantly associated with loin eye area and total lipid in individuals from the BY family. Significant associations were also found between Bsl I polymorphism in LXRA and boneless loin (%), as well as marbling score in one commercial line. The LXRB Aci I polymorphism was significantly associated with lean meat and fat content in the BY family and a number of the commercial lines examined. Our current findings suggested that LXRA and LXRB might have potential effects, especially for loin lean and fat content.