Single nucleotide polymorphism scanning and expression of the FRZB gene in pig populations

Secreted frizzled-related protein 3 (sFRP3), encoded by the gene FRZB, is a member of the sFRP family with important roles in inhibition of the Wnt signalling pathway through competitive binding of the Wnt receptor. Here, we investigated pig FRZB as a candidate gene for growth traits and identified three polymorphic sites, an insertion (A-532B) and two SNPs (G636A and C650T) in its 5′-UTR. The genotype distributions of G636A and C650T were significantly different among mini-type indigenous (Diannan Small-ear and Tibetan), normal indigenous (Laiwu and Huai), and introduced (Large Yorkshire and Landrace) breeds. In semi-quantitative PCR expression analysis, expression of FRZB mRNA was abundant in tissues of hypophysis, longissimus dorsi muscle, and adipose tissues, and low in the heart, hypothalamus, and brain. Quantitative determination of mRNA level and protein expression analysis were corresponding. The results demonstrated that FRZB gene expression in longissimus dorsi muscle and liver tissue was significantly higher in Diannan Small-ear and Tibetan pigs than in the Large Yorkshire breed (P < 0.05); however, in back fat tissue, the expression was significantly higher in Diannan Small-ear pig than in Tibetan or Large Yorkshire breeds (P < 0.05). Given the known growth and fat characteristics of the breeds, these results indicate that FRZB expression has a negative association with muscle growth and a positive association with fat deposition. In conclusion, FRZB may be a major candidate gene for growth traits in pigs.     

Lipogenic enzyme activities in subcutaneous adipose tissue and skeletal muscle from neonatal pigs consuming maternal or formula milk

The influence of maternal and formula milk on lipid metabolism was studied in 7-day-old pigs. Lipid content, fatty acid composition, lipogenic enzyme activities and expression of GLUT4 mRNA were determined in subcutaneous adipose tissue and skeletal muscle from pigs that were bottle-fed formula milk (F) or sow milk (SM), or were sow-reared (SR). Bottle-fed pigs were isoenergetically fed and achieved similar daily body weight gain. SR pigs have a higher (P < 0.05) body weight gain than bottle-fed pigs. Lipid content of adipose tissue was lower (P < 0.05) in F than in SM and SR pigs. In muscle, lipid content did not differ significantly between groups. In adipose tissue, acetyl-CoA-carboxylase (CBX), fatty acid synthase (FAS), malic enzyme (ME), glucose-6-phosphate-dehydrogenase (G6PDH) and lipoprotein lipase (LPL) activities and GLUT4 mRNA levels were higher (P < 0.05) in SR than in bottle-fed pigs. In muscle, ME and G6PDH activities and GLUT4 mRNA were higher (P < 0.05) in F than in SM and SR pigs; LPL was not detected. The present study indicates that lipogenic enzyme activities and GLUT4 mRNA expression are regulated differently in subcutaneous adipose tissue and skeletal muscle in the neonatal pig.     

Changes in the expression of uncoupling proteins and lipases in porcine adipose tissue and skeletal muscle during feed deprivation*(1)

The hormone-sensitive and lipoprotein lipases are critical determinants of the metabolic adaptation to starvation. Additionally, the uncoupling proteins have emerged with potential roles in the metabolic adaptations required by energy deficiency. The objective of this study was to evaluate the expression (mRNA abundance) of uncoupling proteins 2 and 3 and that of hormone-sensitive and lipoprotein lipase in the adipose tissue and skeletal muscle of the pig in relationship to feed deprivation. Thirty-two male castrates (87 kg +/- 5%) were assigned at random to fed and feed-deprived treatment groups. After 96 hr, the pigs were euthanized and adipose and skeletal muscle tissue obtained for total RNA extraction and nuclease protection assays. Feed deprivation increased uncoupling protein 3 mRNA abundance 103-237% (P < 0.01) in longissimus and red and white semitendinosus muscle. In contrast, the increase in uncoupling protein 3 mRNA in adipose tissue was only 23% (P < 0.06), and adipose uncoupling protein 2 mRNA was not influenced (P > 0.66) by feed deprivation. The increased abundance of uncoupling protein 2 mRNA in the longissimus muscle of feed-deprived pigs was small (22%), but significant (P < 0.04). The expression of hormone-sensitive lipase was increased 46% and 64% (P < 0.04) in adipose tissue and longissimus muscle, respectively, by feed deprivation, whereas adipose lipoprotein lipase expression was reduced (P < 0.01) to 20% of that of the fed group. Longissimus lipoprotein lipase expression in the feed-deprived group was 37% of that of the fed group (P < 0.01), and similar reductions were detected in red and white semitendinosus muscle. Overall, these findings indicate that uncoupling protein 3 expression in skeletal muscle is quite sensitive to starvation in the pig, whereas uncoupling protein 2 changes are minimal. Furthermore, we conclude that hormone-sensitive lipase is upregulated at the mRNA level with prolonged feed deprivation, whereas lipoprotein lipase is downregulated.     

Expression profiling analysis for genes related to meat quality and carcass traits during postnatal development of backfat in two pig breeds

The competitive equilibrium of fatty acid biosynthesis and oxidation in vivo determines porcine sub-cutaneous fat thickness(SFT) and intramuscular fat(IMF) content.Obese and lean-type pig breeds show obvious differences in adipose deposition;however, the molecular mechanism underlying this phenotypic variation remains unclear.We used pathway-focused oligo microarray studies to examine the expression changes of 140 genes associated with meat quality and carcass traits in backfat at five growth stages(1―5 months) of Landrace(a leaner, Western breed) and Taihu pigs(a fatty, indigenous, Chinese breed).Variance analysis(ANOVA) revealed that differences in the expression of 25 genes in Landrace pigs were significant(FDR adjusted permutation, P<0.05) among 5 growth stages.Gene class test(GCT) indicated that a gene-group was very significant between 2 pig breeds across 5 growth stages(PErmineJ<0.01), which consisted of 23 genes encoding enzymes and regulatory proteins associ-ated with lipid and steroid metabolism.These findings suggest that the distinct differences in fat deposition ability between Landrace and Taihu pigs may closely correlate with the expression changes of these genes.Clustering analysis revealed a very high level of significance(FDR adjusted, P<0.01) for 2 gene expression patterns in Landrace pigs and a high level of significance(FDR adjusted, P<0.05) for 2 gene expression patterns in Taihu pigs.Also, expression patterns of genes were more diversified in Taihu pigs than those in Landrace pigs, which suggests that the regulatory mechanism of micro-effect polygenes in adipocytes may be more complex in Taihu pigs than in Landrace pigs.Based on a dy-namic Bayesian network(DBN) model, gene regulatory networks(GRNs) were reconstructed from time-series data for each pig breed.These two GRNs initially revealed the distinct differences in physiological and biochemical aspects of adipose metabolism between the two pig breeds;from these results, some potential key genes could be identified.Quantitative, real-time RT-PCR(QRT-PCR) was used to verify the microarray data for five modulated genes, and a good correlation between the two measures of expression was observed for both 2 pig breeds at different growth stages(R=0.874±0.071).These results highlight some possible candidate genes for porcine fat characteristics and provide some data on which to base further study of the molecular basis of adipose metabolism.     

The association of haplotypes in IGFBP-3 gene promoter region and tissue expressions in three pig breeds

Bama Xiang pig (BM) and Tibetan mini-pig (TM) are used as experimental animals in China; however, the dwarf molecular mechanisms of these Chinese local pig breeds are unknown. IGFBP-3 affects animal growth, carcass and meat quality. The aim of this study was to identify the polymorphisms in the promoter of the IGFBP-3 and analyse their effect on the IGFBP-3 mRNA expression level in liver and muscle tissues. High-density single-nucleotide polymorphisms (SNPs) (31) and InDels (5) were detected in the promoter of the IGFBP-3 in the BM, TM and Junmu No. 1 White (JM, control) pig breeds from 114 individuals by re-sequencing. A perfect Linkage disequilibrium consisted of 13 SNPs was observed in the promoter region and 2 main haplotypes were identified, of which the h1 genotype (GCA-ATGTACATAT) was more prevalent in JM breed than in TM or BM breeds (P?h2 (ATGTGCACG--CGC) was the dominant haplotype in TM and BM breeds (P?IGFBP-3 mRNA expression level in liver and muscle tissues of pigs. The IGFBP-3 mRNA expression level was determined higher in the liver and muscle tissues of pigs with h2 genotype as compared to that in pigs with h1 genotype (P?IGFBP-3 gene may serve as useful molecular markers for the body growth traits and the breeding in swine.     

Effect of simvastatin on culturing of kidney cells from pigs in vitro

Statins is an inhibitor in the cholesterol biosynthesis pathway. Simvastatin (SIM) has been found to have other clinical benefits besides those resulted from its actions of reducing plasma level of low density lipoprotein (LDL) cholesterol. Both mevastatin (MEV) and parvastatin (PAR) can increase release of nitric oxide (NO) which is a potent inhibitor of platelet aggregation and endothelial cell conglutination. In this study, we found different concentrations of SIM had different effects on 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) mRNA expression and NO and total cholesterol (TC) in normal cultural pig kidney cells. NO and TC were measured by using colorimetry in 550 nm and 546 nm, respectively. HMGR mRNA expression was tested by RT-PCR. Results showed that HMGR mRNA expression had a significant difference (P < 0.05) between different concentration of SIM treatment (0, 5, 10, or 25 μmol/l). HMGR mRNA expression and TC content decreased gradually with the elevation of SIM concentration. The content of NO increased with the elevation of SIM concentration, but the difference was not notable. SIM affects the expression of HMGR-CoA, TC and NO in normal cells, but the specific mechanism need to be further research.     

Expression profiling analysis for genes related to meat quality and carcass traits during postnatal development of backfat in two pig breeds

The competitive equilibrium of fatty acid biosynthesis and oxidation in vivo determines porcine subcutaneous fat thickness (SFT) and intramuscular fat (IMF) content. Obese and lean-type pig breeds show obvious differences in adipose deposition; however, the molecular mechanism underlying this phenotypic variation remains unclear. We used pathway-focused oligo microarray studies to examine the expression changes of 140 genes associated with meat quality and carcass traits in backfat at five growth stages (1–5 months) of Landrace (a leaner, Western breed) and Taihu pigs (a fatty, indigenous, Chinese breed). Variance analysis (ANOVA) revealed that differences in the expression of 25 genes in Landrace pigs were significant (FDR adjusted permutation, P<0.05) among 5 growth stages. Gene class test (GCT) indicated that a gene-group was very significant between 2 pig breeds across 5 growth stages (P _ErmineJ<0.01), which consisted of 23 genes encoding enzymes and regulatory proteins associated with lipid and steroid metabolism. These findings suggest that the distinct differences in fat deposition ability between Landrace and Taihu pigs may closely correlate with the expression changes of these genes. Clustering analysis revealed a very high level of significance (FDR adjusted, P<0.01) for 2 gene expression patterns in Landrace pigs and a high level of significance (FDR adjusted, P<0.05) for 2 gene expression patterns in Taihu pigs. Also, expression patterns of genes were more diversified in Taihu pigs than those in Landrace pigs, which suggests that the regulatory mechanism of micro-effect polygenes in adipocytes may be more complex in Taihu pigs than in Landrace pigs. Based on a dynamic Bayesian network (DBN) model, gene regulatory networks (GRNs) were reconstructed from time-series data for each pig breed. These two GRNs initially revealed the distinct differences in physiological and biochemical aspects of adipose metabolism between the two pig breeds; from these results, some potential key genes could be identified. Quantitative, real-time RT-PCR (QRT-PCR) was used to verify the microarray data for five modulated genes, and a good correlation between the two measures of expression was observed for both 2 pig breeds at different growth stages (R=0.874±0.071). These results highlight some possible candidate genes for porcine fat characteristics and provide some data on which to base further study of the molecular basis of adipose metabolism.     

猪PID1基因CDS区的克隆及其mRNA表达与肌内脂肪沉积关系

为了探索PID1(Phosphotyrosine interaction domain containing1)基因的表达与脂肪沉积的关系,文章利用兼并引物进行RT-PCR从猪脂肪和肌肉组织中克隆PID1基因CDS(Coding region)区全序列,并采用荧光定量PCR方法对大白猪、鲁莱黑猪、莱芜猪3个猪品种的肝脏、脂肪和肌肉组织PID1基因mRNA表达进行了相对定量分析。结果表明:经克隆、测序,得到了猪PID1基因654bp全编码区序列,通过Blast比对,与人、大鼠、牛有93.88%、66.94%、88.07%的同源性。PID1基因在同一个品种猪中mRNA表达水平总体表现为:肝脏脂肪肌肉。在不同品种3种组织中PID1基因mRNA表达水平总体表现为:莱芜猪鲁莱黑猪大白猪,其中肝脏中差异显著(P0.05),但是在脂肪和肌肉组织中莱芜猪与鲁莱黑猪差异不显著(P0.05)。对于高肌内脂肪(LWH)、中等肌内脂肪(LWI)和低肌内脂肪(LWL)沉积的3组莱芜猪,PID1基因在肝脏组织中的表达水平是LWH显著高于LWL(P0.05),在肌肉组织中则是LWH显著高于LWI和LWL(P0.05)。PID1基因在莱芜猪品种内3个组织中mRNA表达量与IMF含量相关均不显著,而在品种间3个组织中mRNA表达量与IMF含量呈显著正相关(P0.05)。结果提示:PID1的表达可能与脂肪沉积性状存在一定的关系。     

Breed difference and regulation of the porcine adipose triglyceride lipase and hormone sensitive lipase by TNFα

Adipose triglyceride lipase (ATGL) and hormone sensitive lipase (HSL) are major novel triglyceride lipases in animals. The aim of this study was to determine if there are differences in the porcine ATGL ( pATGL ) and HSL genes between Jinhua pigs (a fatty breed) and Landrace pigs (a leaner breed). In addition, the effect of TNFα and pATGL-specific siRNA ( pATGL-siRNA ) on the expression of pATGL and HSL in porcine adipocytes was also examined. Compared with Landrace pigs, the body weight ( BW ) of Jinhua pigs was lower ( P <  0.01), while intramuscular fat content (in the longissimus dorsi muscle), as well as the back fat thickness and body fat content were higher ( P <  0.01). The expression of pATGL and HSL mRNA in Jinhua pigs was lower ( P <  0.01) in subcutaneous adipose tissue, and greater ( P <  0.01) in longissimus dorsi muscle compared with Landrace pigs. In vitro treatment of porcine adipocytes with TNFα decreased ( P <  0.01) the glycerol release and the gene expression of pATGL , HSL and PPARγ in porcine adipocytes. Furthermore, transfection with pATGL-siRNA significantly decreased ( P <  0.01) the expression of pATGL , while it had no effect on the expression of HSL . Treatment with 25 ng/ml TNFα in conjunction with pATGL-siRNA significantly decreased ( P <  0.01) the expression of pATGL and HSL in cultured porcine adipocytes. These results provide useful information to further the understanding of the function of pATGL and HSL in porcine lipid metabolism, which should be applicable to the regulation of fat deposition and improvement of meat quality.     

猪脂肪组织中IGF2和IGFBP3基因表达的发育性变化及其品种差异

采用荧光定量PCR技术分析长白猪和太湖猪背部皮下脂肪组织中胰岛素样生长因子2和胰岛素样生长因子结合蛋白3基因在30、60、90、120和150日龄时表达水平的发育性变化。结果表明: (1)品种内日龄间比较, 长白猪IGF2 mRNA 在30日龄时的表达量极显著高于其他日龄(P<0.01), 之后逐渐下降, 至120日龄降到最低, 150日龄时又明显上升; 太湖猪IGF2 mRNA在30~60日龄的表达量较高, 90日龄降至最低, 120日龄迅速回升, 之后又有所下降。两品种IGFBP3 mRNA表达的发育性变化模式基本相同, 30日龄时表达量最高, 60日龄显著下降(P<0.05), 之后趋于平缓但略有波动。(2)品种间同日龄比较, 120日龄时太湖猪IGF2 mRNA的表达量极显著高于长白猪(P<0.01), 150日龄时太湖猪IGFBP3的表达量极显著低于长白猪(P<0.01), 其余日龄间两品种差异均不显著(P>0.05)。研究结果提示: 猪脂肪组织IGF2和IGFBP3基因表达存在明显的发育性变化和品种差异; IGF2基因表达水平的降低可能与脂肪细胞的增殖有关。     

缺乏抗坏血酸对豚鼠胆固醇代谢的影响

边缘性缺乏抗坏血酸之豚鼠,于三周内其肝脏及小肠粘膜3-羟-3-甲基戊二酰辅酶A还原酶(HMGR)活力均下降到原有水平的50％,但肝脏胆固醇7α-羟化酶活力尚无显著性改变。坏血病豚鼠(三周内)上述几种酶活力都下降至原有水平的50％左右。豚鼠摄取抗坏血酸不足,其血清总胆固醇浓度显著增加,而血清高密度脂蛋自胆固醇浓度显著减少,其改变程度与抗坏血酸缺乏状况一致。     

猪背最长肌中肌肉生长和脂肪沉积相关基因的差异表达和主成分分析

骨骼肌细胞和脂肪细胞在分化生长速度上相对竞争的平衡点是猪肉质和胴体性状的决定因素.利用Oligo功能分类芯片检测了瘦肉型的长白猪和脂肪型的太湖猪在初生、1、2、3、4和5月龄间背最长肌中肌肉生长和脂肪沉积相关基因的动态表达变化.差异表达分析结果显示,在初生至5月龄的品种间分别有15、16、11、13、18和20个基因的表达差异倍数大于2倍.品种内的方差分析表明,长白猪分别有18和22个基因,太湖猪分别有3和7个基因在月龄间的表达差异达极显著(Ｐ<0.01)和显著水平(Ｐ<0.05).主成分分析结果显示,先降后升是两品种内最具代表性的基因表达模式,且长白猪和太湖猪分别有7和6个基因的表达模式明显偏离其他基因,提示其可能受到了重要的调控. 此外,5个差异表达基因的荧光定量RT-PCR验证结果均与芯片结果呈正相关趋势.以上结果筛选出了对于猪肉质和胴体性状可能具有重要影响,值得深入研究的一些候选基因,为深入研究生长发育过程中参与肌纤维生长和脂肪酸合成关键基因的表达变化规律和互作调控机制提供了基础数据.     

Developmental patterns of serum leptin levels, leptin gene expression in adipose tissue and Ob-Rb gene expression in hypothalamus of Erhualian and Large White pigs

~~Developmental patterns of serum leptin levels, leptin gene expression in adipose tissue and Ob-Rb gene expression in hypothalamus of Erhualian and Large White pigs~~     

Clofibrate causes an upregulation of PPAR-{alpha} target genes but does not alter expression of SREBP target genes in liver and adipose tissue of pigs

This study investigated the effect of clofibrate treatment on expression of target genes of peroxisome proliferator-activated receptor (PPAR)-alpha and various genes of the lipid metabolism in liver and adipose tissue of pigs. An experiment with 18 pigs was performed in which pigs were fed either a control diet or the same diet supplemented with 5 g clofibrate/kg for 28 days. Pigs treated with clofibrate had heavier livers, moderately increased mRNA concentrations of various PPAR-alpha target genes in liver and adipose tissue, a higher concentration of 3-hydroxybutyrate, and markedly lower concentrations of triglycerides and cholesterol in plasma and lipoproteins than control pigs (P < 0.05). mRNA concentrations of sterol regulatory element-binding proteins (SREBP)-1 and -2, insulin-induced genes (Insig)-1 and Insig-2, and the SREBP target genes acetyl-CoA carboxylase, 3-methyl-3-hydroxyglutaryl-CoA reductase, and low-density lipoprotein receptor in liver and adipose tissue and mRNA concentrations of apolipoproteins A-I, A-II, and C-III in the liver were not different between both groups of pigs. In conclusion, this study shows that clofibrate treatment activates PPAR-alpha in liver and adipose tissue and has a strong hypotriglyceridemic and hypocholesterolemic effect in pigs. The finding that mRNA concentrations of some proteins responsible for the hypolipidemic action of fibrates in humans were not altered suggests that there were certain differences in the mode of action compared with humans. It is also shown that PPAR-alpha activation by clofibrate does not affect hepatic expression of SREBP target genes involved in synthesis of triglycerides and cholesterol homeostasis in liver and adipose tissue of pigs.     

Functional and association studies of the cholesteryl ester transfer protein (CETP) gene in a Wannan Black pig model

Some polymorphisms of the human CETP gene are causally and significantly associated with serum lipids levels; however, the information regarding this gene in pigs is sparse. To evaluate the effects of CETP on blood lipid traits and fat deposition in pig, porcine CETP tissue expression patterns were observed by quantitative real‐time polymerase chain reaction (qPCR) first. High expression was detected in liver, spleen, gluteus medius (GM) muscle and backfat. A de novo polymorphism (AF333037:g.795C>T) in the intron 1 region of porcine CETP was identified. This polymorphism was further genotyped by direct sequencing of the PCR products of 390 Wannan Black pigs, a Chinese native breed population. Association analyses at 45 and 300 days of age revealed highly significant associations between CETP genotypes and serum lipid traits. Furthermore, this polymorphism was proved to be associated with differences in liver CETP mRNA levels: pigs at 300 days of age with the TT genotype had higher levels than did those with other genotypes (P = 0.021). Additionally, analysis at 300 days of age showed that GM CETP mRNA expression correlated positively with serum lipids levels as well as with carcass backfat thickness and intramuscular fat content in GM. These results indicate that CETP is involved in serum, adipose and muscle lipid metabolism in pigs. The mechanisms underlying such relationships and their functional implications are worthy of further research.     

Fat metabolism is regulated by altered gene expression of lipogenic enzymes and regulatory factors in liver and adipose tissue but not in semimembranosus muscle of pigs during the fattening period

It has been shown previously that lipid metabolism is regulated by fatty acids (FA) and that thyroid hormones are important regulators of energy metabolism. The effects of weight, dietary fat level and dietary FA profile on thyroid hormone levels and expression of lipogenic genes and tissue FA composition were studied. Sixty-one crossbred gilts weighing 62 ± 5.2 kg BW average were either slaughtered at the beginning of the trial (n = 5) or fed one of seven diets (n = 8 pigs per diet): a semi-synthetic diet formulated to contain a very low level of fat (NF) and six diets based on barley-soybean meal supplemented with approximately 10% fat of different origin and slaughtered at 100 kg BW. The supplemental fats were tallow, high-oleic sunflower oil, sunflower oil (SFO), linseed oil, fat blend (55% tallow, 35% sunflower oil, 10% linseed oil) and fish oil blend (40% fish oil, 60% linseed oil). In general, the dietary FA profiles altered the FA composition of liver, semimembranosus muscle and adipose tissues. Pigs fed the NF diet had the highest free and total triiodothyronine (T3) values followed by pigs fed SFO. Total T3 levels were higher in pigs at 60 kg than in pigs at 100 kg. Correlations between thyroid hormones and genes encoding enzymes of fat synthesis in adipose tissue (acetyl CoA carboxylase (ACACA), fatty acid synthase and stearoyl CoA desaturase (SCD)) and the large differences in expression of lipogenic genes at different weights (60 and 100 kg BW), suggest a role for thyroid hormones and for T3, in particular, in regulating whole animal fat metabolism, with effects brought about by altered expression of lipogenic genes. Liver sterol receptor element binding protein-1 (SREBP1) mRNA content was affected by dietary treatment (P < 0.001) and was correlated with ACACA and SCD, whereas adipose tissue SREBP1 was not correlated with the mRNA abundance of any lipogenic enzyme. Weight and tissue factors showed greater influence on mRNA abundance of genes related with lipid metabolism than diet and tissue FA composition. In the pig, FA synthesis appear to be of greater magnitude in adipose tissue than in the liver as suggested by the higher expression of lipogenic genes in adipose tissue.     

Dietary fat source affects metabolism of fatty acids in pigs as evaluated by altered expression of lipogenic genes in liver and adipose tissues

Little is known about pig gene expressions related to dietary fatty acids (FAs) and most work have been conducted in rodents. The aim of this study was to investigate how dietary fats regulate fat metabolism of pigs in different tissues. Fifty-six crossbred gilts (62 ± 5.2 kg BW) were fed one of seven dietary treatments (eight animals per treatment): a semi-synthetic diet containing a very low level of fat (no fat (NF)) and six fat-supplemented diets (ca. 10%) based on barley and soybean meal. The supplemental fat sources were tallow (T), high-oleic sunflower oil (HOSF), sunflower oil (SFO), linseed oil (LO), blend (FB) (55% T, 35% SFO and 10% LO) and fish oil (FO) blend (40% FO and 60% LO). Pigs were slaughtered at 100 kg BW and autopsies from liver, adipose tissue and muscle semimembranousus were collected for qPCR. The messenger ribonucleic acid (mRNA) abundances of genes related to lipogenesis were modified due to dietary treatments in both liver (sterol regulatory element-binding protein-1 (SREBP-1), acetyl CoA carboxylase (ACACA) and stearoyl CoA desaturase (SCD)) and adipose tissue (fatty acid synthase (FASN), ACACA and SCD), but were not affected in semimembranousus muscle. In the liver, the mRNA abundances of genes encoding lipogenic enzymes were highest in pigs fed HOSF and lowest in pigs fed FO. In adipose tissue, the mRNA abundances were highest in pigs fed the NF diet and lowest in pigs fed T. The study demonstrated that dietary FAs stimulate lipogenic enzyme gene expression differently in liver, fat and muscles tissues.     

Characterization of <Emphasis Type="Italic">methionine adenosyltransferase 2β</Emphasis> gene expression in skeletal muscle and subcutaneous adipose tissue from obese and lean pigs

Methionine adenosyltransferase (MAT) catalyzes the biosynthesis of S-adenosylmethionine. Two genes (MAT1A and MAT2A) encode for the catalytic subunit of MAT, while a third gene (MAT2β) encodes for a regulatory subunit (MAT II β) that regulates the activity of the MAT2A-encoded isoenzyme and intracellular S-adenosylmethionine levels. Our previous work identified MAT2β as a candidate gene for intramuscular fat (IMF) deposition in porcine skeletal muscle by microarray technology. Here, we cloned porcine MAT2β cDNA and compared its expression pattern in subcutaneous adipose tissue and skeletal muscle from obese (Rongchang Breed) and lean (Pig Improvement Company, PIC) pigs (n = 6). The porcine MAT2β cDNA was 1,800 bp long and encodes for 334 amino acids sharing high similarity with other species. MAT2β is expressed at a higher level in liver and duodenum, followed by the stomach, fat and longissinus dorsi muscle. As expected, both subcutaneous fat content and IMF content were higher in obese than in lean pigs (both P < 0.01). MAT2β mRNA abundance was lower in both subcutaneous adipose tissue and skeletal muscle in obese pigs compared with lean pigs (both P < 0.01). MAT II β protein content was lower in skeletal muscle in obese than in lean pigs (P < 0.05), whereas the opposite was observed in subcutaneous adipose tissue (P < 0.01). These data demonstrated an obesity-related expression variation of the MAT II β subunit in skeletal muscle and adipose tissue in pigs, and suggest a novel role for the MAT2β gene in regulation of IMF deposition in skeletal muscle.     

Chronic leptin administration increases serum NEFA in the pig and differentially regulates PPAR expression in adipose tissue

Two in vivo studies were conducted with pigs to determine the effects of exogenous leptin on the expression of peroxisome proliferator activated receptors (PPAR), and on serum concentrations of selected metabolites and hormones. Initially, leptin was administered i.m. to young pigs for 15 days at 0 (control), 0.003 (low), 0.01 (medium) and 0.03 (high) mg. kg(-1). day(-1). There was no leptin effect on serum glucose (P > 0.84), triglycerides (P > 0.69), non-esterified fatty acids (NEFA, P > 0.53), or glycerol (P > 0.33). Leptin at the intermediate and high doses depressed adipose expression of both PPARgamma1 (P < 0.06) and PPARgamma2 (P < 0.01). In a second study, we used a paired-feeding experimental design to determine the effects of a higher dose of leptin (0.05 mg. kg(-1). day(-1)) on serum metabolites and PPAR expression in selected tissues. At this dose, leptin increased (P < 0.0001) serum NEFA concentrations relative to both the ad libitum and pair-fed control groups. However, in this study, there was no difference in the expression of PPARgamma1 in adipose tissue, but PPARgamma2 mRNA was upregulated by leptin (P < 0.08). In contrast, leptin had no impact on the expression of PPARalpha in liver, skeletal muscle or adipose tissue. Adipose tissue explants were also incubated with leptin to assess the effect on PPARgamma expression, in vitro. The abundance of PPARgamma1 mRNA (P < 0.05) was increased after 24 hr of exposure, but the effect of leptin on gamma2 was not significant (P > 0.24). The lipolytic effect of leptin was also evaluated in vitro using isolated adipocytes. In keeping with the increase in serum NEFA concentrations in vivo, leptin stimulated lipolysis in vitro, increasing glycerol concentrations in the medium to about 219% of that in basal (non-treated) culture medium after 8 hr of incubation. Collectively, the data presented herein indicate that leptin modulates lipid metabolism in the pig, but that PPARalpha expression is not a parallel target of leptin as it is in rodent models. The regulation of PPARgamma by leptin seems complex in that it varied in relation to dose in vivo, and may be impacted by in vitro vs. in vivo circumstances.