Characterization of key genes of the renin–angiotensin system in mature feline adipocytes and during in vitro adipogenesis

Obesity is a growing health problem in humans as well as companion animals. In the development and progression of obesity‐associated diseases, the members of the renin–angiotensin system (RAS) are proposed to be involved. Particularly, the prevalence of type 2 diabetes mellitus in cats has increased enormously which is often been linked to obesity as well as to RAS. So far, reports about the expression of a local RAS in cat adipocytes are missing. Therefore, we investigated the mRNA expression of various RAS genes as well as the adipocyte marker genes adiponectin, leptin and PPAR‐γ in feline adipocytes using quantitative PCR. To characterize the gene expression during adipogenesis, feline pre‐adipocytes were differentiated into adipocytes in a primary cell culture and the expression of RAS key genes measured. All major RAS components were expressed in feline cells, but obvious differences in the expression between pre‐adipocytes and the various differentiation stages were found. Interestingly, the two enzymes ACE and ACE2 showed an opposite expression course. In addition to the in vitro experiments, mature adipocytes were isolated from subcutaneous and visceral adipose tissue. Significant differences between both fat depots were found for ACE as well as AT1 receptor with greater expression in subcutaneous than in visceral adipocytes. Visceral adipocytes had significantly higher adiponectin and PPAR‐γ mRNA level compared to the subcutaneous fat cells. Concerning the nutritional status, a significant lower expression of ACE2 was measured in subcutaneous adipocytes of overweight cats. In summary, the results show the existence of a potentially functional local RAS in feline adipose tissue which is differentially regulated during adipogenesis and dependent on the fat tissue depot and nutritional status. These findings are relevant for understanding the development of obesity‐associated diseases in cats such as diabetes mellitus.     

Adipogenic differentiation state-specific gene expression as related to bovine carcass adiposity

Genetic regulation of the site of fat deposition is not well defined. The objective of this study was to investigate adipogenic differentiation state-specific gene expression in feedlot cattle (>75% Angus; <25% Simmental parentage) of varying adipose accretion patterns. Four groups of 4 steers were selected via ultrasound for the following adipose tissue characteristics: low subcutaneous-low intramuscular (LSQ-LIM), low subcutaneous-high intramuscular (LSQ-HIM), high subcutaneous-low intramuscular (HSQ-LIM), and high subcutaneous-high intramuscular (HSQ-HIM). Adipose tissue from the subcutaneous (SQ) and intramuscular (IM) depots was collected at slaughter. The relative expression of adipogenic genes was evaluated using quantitative PCR. Data were analyzed using the mixed model of SAS, and gene expression data were analyzed using covariate analysis with ribosomal protein L19 as the covariate. No interactions (P > 0.10) were observed between IM and SQ adipose tissue depots for any of the variables measured. Therefore, only the main effects of high and low accretion within a depot and the effects of depot are reported. Steers with LIM had smaller mean diameter IM adipocytes (P < 0.001) than HIM steers. Steers with HSQ had larger mean diameter SQ adipocytes (P < 0.001) than LSQ. However, there were no differences (P > 0.10) in any of the genes measured due to high or low adipose accretion. Preadipogenic delta-like kinase1 mRNA was greater in the IM than the SQ adipose tissue; conversely, differentiating and adipogenic genes, lipoprotein lipase, PPARγ, fatty acid synthetase, and fatty acid binding protein 4 were greater (P < 0.001) in the SQ than the IM depot. Intramuscular adipocytes were smaller than SQ adipocytes and had greater expression of the preadipogenic gene, indicating that more hyperplasia was occurring. Meanwhile, SQ adipose tissue contained much larger (P < 0.001) adipocytes that had a greater expression (P < 0.001) of differentiating and adipogenic genes than did the IM adipose tissue, indicating more cells were undergoing differentiation and hypertrophy. Adipogenic differentiation state-specific gene expression was not different in cattle with various phenotypes, but adipogenesis in the SQ and IM adipose tissues seems to occur independently.     

Bovine haptoglobin as an adipokine: serum concentrations and tissue expression in dairy cows receiving a conjugated linoleic acids supplement throughout lactation

The present study aimed to characterize serum haptoglobin (Hp) concentrations throughout an entire lactation period in both primi- and multiparous cows and to compare them to the Hp mRNA expression in liver and - in view of Hp being potentially an adipokine - also in different subcutaneous (s.c.) and visceral fat depots. In addition, potential anti-inflammatory effects of long-term supplementation with conjugated linoleic acids (CLA) were evaluated by assessing Hp. Trial 1 comprised 33 cows and 16 Holstein heifers from day 21 ante partum until day 252 postpartum. The animals received 100 or 50 g/day CLA or a control fat supplement. Blood samples and biopsy (tail head fat and liver) samples were collected. Trial 2 included 25 Holstein heifers, 5 animals were slaughtered on the day of parturition, the remaining animals were allocated to either CLA (100 g/day, n=10) or control fat supplement (n=10) and slaughtered on days 42 and 105 postpartum, respectively. At slaughter, fat samples were collected from 3 different visceral depots, 3 s.c. depots and from liver tissue. Results indicated no effects of CLA on serum Hp and liver Hp mRNA for both trials and on Hp mRNA in biopsies from s.c. tail head fat. In omental and s.c. withers fat from trial 2, CLA reduced Hp mRNA on both day 42 and day 105. Hp mRNA was detectable in fat tissues from both trials with abundance values being significantly lower than in liver. The Hp mRNA abundance in the s.c. fat depots was generally higher than in the visceral depots. Haptoglobin mRNA abundance in the different tissues from trial 2 was correlated whereby all s.c. depots were interrelated. The evidence of Hp mRNA expression in adipose tissues and the presence of Hp-immune staining in histological fat sections confirm that Hp can be classified as a bovine adipokine. The lack of an evident relationship between circulating Hp concentrations and normal body fat portions in dairy cattle demonstrates that varying degrees of adiposity are not confounding factors when using Hp as inflammatory marker. The physiological changes in serum Hp concentration seem to be limited to parity and parturition. In view of the lack of effects of CLA on serum Hp concentrations, the observed reaction in two out of six different fat depots seems of marginal importance for the organisms as an entity.     

Adipose tissue growth and cellularity: changes in bovine adipocyte size and number

Forty crossbred steers of similar birth date and fed the same growing-finishing diet were used to study adipocyte changes in six fat depots during growth from 11 to 19 mo of age. Steers were slaughtered at 2-mo intervals. Adipose tissue samples were obtained from kidney, mesenteric and brisket fat and subcutaneous, intermuscular and intramuscular fat from the 10th to 12th rib section. The osmium tetroxide fixation technique was used for determination of cell size and number. Except for three brisket fat samples, distributions of adipocyte diameters from six different fat depots were monophasic during the age range considered in this study. At 17 mo of age, the mean adipocyte diameter, in decreasing order, was: kidney fat greater than mesenteric greater than subcutaneous greater than intermuscular greater than intramuscular greater than brisket fat. Fat deposition during growth to 19 mo of age occurred mainly by hypertrophy of adipocytes. An apparent cell hyperplasia occurred in the intramuscular fat depot from 11 to 15 mo and in the brisket fat depot after 15 mo of age. Based on cellularity characteristics, evidence exists to classify intramuscular and brisket fat depots as late-developing ones. Cell number/gram of intramuscular adipose tissue was a better predictor of marbling score than was fat cell diameter.     

Expression of adiponectin and its receptors in swine

Adiponectin is an adipocyte-derived hormone that plays an important role in lipid metabolism and glucose homeostasis. Objectives of this study were 1) to determine the presence and distribution of adiponectin and its receptors 1 and 2 (adipoR1 and adipoR2) in porcine tissues; 2) to characterize pig adiponectin, adipoR1, and adipoR2 mRNA levels in various fat depots from three different breeds of pigs; and 3) to study, in stromal-vascular cell culture, the effects of leptin and tumor necrosis factor-alpha (TNFalpha) on pig adiponectin, adipoR1, and adipoR2 gene expression. To this end, fat Chinese Upton Meishan (UM, n = 10), lean Ham Line (HL, n = 10), and Large White (LW, n = 10) gilts were used. We report the isolation of partial cDNA sequences of pig adipoR1 and adipoR2. Porcine-deduced AA sequences share 97 to 100% homology with human and murine sequences. Pig adipoR1 mRNA is abundant in skeletal muscle, visceral fat, and s.c. fat tissues, whereas adipoR2 mRNA is predominantly expressed in liver, heart, skeletal muscle, and visceral and s.c. fat tissues. Pig adiponectin mRNA levels in s.c. and visceral fat tissues were not associated with plasma insulin and glucose in fasting animals. Subcutaneous (r = -0.44, P < 0.05), visceral (r = -0.43, P < 0.05), and total body fat (r = -0.42, P < 0.05) weights were negatively correlated with adiponectin mRNA levels measured in visceral, but not s.c., fat. Pig adipoR1 and adipoR2 mRNA levels, in visceral fat, were less expressed in fat UM gilts than in the lean HL gilts (P < 0.05). Inverse associations were found between s.c. (r = -0.57, P < 0.01), visceral (r = -0.46, P < 0.05), and total body fat (r = -0.56, P < 0.01) weights and adipoR2 mRNA levels in visceral fat only. We were unable to find such associations for adipoR1 mRNA levels in the overall gilt population. The current study demonstrated that TNFalpha downregulates adiponectin and adipoR2, but not adi-poR1, mRNA levels in stromal-vascular cell culture. Moreover, leptin significantly decreased adiponectin mRNA levels, whereas there was no effect on adiponectin receptors. We conclude that adiponectin and adi-poR2 mRNA levels, but not adipoR1, are modulated in pig visceral fat tissues. Furthermore, our results indicate that TNFalpha interferes with adiponectin function by downregulation of adipoR2 but not of adipoR1 mRNA levels in pigs.     

ORIGINAL ARTICLE: mRNA abundance of adiponectin and its receptors,leptin and visfatin and of G‐protein coupled receptor 41 in five different fat depots from sheep

Adipose tissue (AT) expresses adipokines, which are involved in the regulation of energy expenditure, lipid metabolism and insulin sensitivity. Visceral (v.c.) and subcutaneous (s.c.) depots largely differ concerning their metabolic characteristics as to the control of lipolysis and the sensitivity to insulin. The adipokines adiponectin, leptin and visfatin influence lipolysis and insulin sensitivity. Signalling by G‐protein coupled receptor 41 (GPR 41) stimulates leptin release via activation by short‐chain fatty acids. We hypothesized that the metabolic differences between v.c. and s.c. fat depots may also apply to the expression of adiponectin, its receptors, leptin, visfatin, insulin receptor (IR) and GPR 41. Therefore, we aimed to compare the mRNA expression of adiponectin, leptin and visfatin, of the adiponectin receptors 1 and 2 (AdipoR1/2) and IR as well of GPR 41 between several s.c. and v.c. fat depots in sheep. Samples from 10 rams were collected at slaughter (40 kg BW) from three s.c. depots, i.e. close to sternum (s.c.S), close to withers (s.c.W), and at the base of tail (s.c.T), and from two v.c. depots, i.e. from perirenal (v.c.P) and omental (v.c.O) fat. The mRNAs of both adiponectin receptors, as well as IR and putative GPR 41, were higher expressed in v.c. fat than in s.c. fat (p ≤ 0.05). Leptin mRNA abundance was greater in s.c. than in v.c. fat (mean ± SEM: s.c.: 2.55 ± 0.81; v.c.: 0.66 ± 0.21) and also differed among the five separately measured fat depots. Our results show differences in mRNA abundance for leptin, AdipoR1 and R2, as well as for IR and GPR 41 in s.c. compared with v.c. fat, thus confirming the need for individual consideration of distinct fat depots, when aiming to characterize adipose functions in ruminants.     

Expression of lipogenic genes during porcine intramuscular preadipocyte differentiation

Intramuscular fat (IMF) content plays an important role in meat quality. Triglyceride (TG) metabolism in intramuscular adipocytes is strongly associated with the intramuscular fat deposition. To better understand the mechanisms leading to IMF deposition we compared the expression levels of genes related to preadipocyte differentiation and lipogenesis in the intramuscular preadipocytes isolated from the longissimus muscle of Wujin and Landrace pigs. The results showed that the intramuscular preadipocytes could differentiate into mature adipocytes in vitro. Triglyceride content in adipocytes isolated from Wujin pigs was higher than Landrace pigs during the middle and later phases of preadipocyte differentiation. The expression levels of genes related to preadipocyte differentiation such as PPARG and CEBPA showed differential expression between Wujin and Landrace porcine adipocytes during the early stage of differentiation. The expression levels of lipogenic genes such as FASN and SREBF1 were significantly higher in Wujin porcine intramuscular preadipocytes than in Landrace intramuscular preadipocytes at the middle and the later stages of differentiation. This suggests that preadipocyte differentiation and lipogenesis exhibited breed-related scheduling.     

11-Hydroxy-β-steroid dehydrogenase gene expression in canine adipose tissue and adipocytes: stimulation by lipopolysaccharide and tumor necrosis factor α

The enzyme 11β-hydroxysteroid dehydrogenase 1 (11β-HSD-1) is expressed in a number of tissues in rodents and humans and is responsible for the reactivation of inert cortisone into cortisol. Its gene expression and activity are increased in white adipose tissue (WAT) from obese humans and may contribute to the adverse metabolic consequences of obesity and the metabolic syndrome. The extent to which 11β-HSD-1 contributes to adipose tissue function in dogs is unknown; the aim of the present study was to examine 11β-HSD-1 gene expression and its regulation by proinflammatory and anti-inflammatory agents in canine adipocytes. Real-time PCR was used to examine the expression of 11β-HSD-1 in canine adipose tissue and canine adipocytes differentiated in culture. The mRNA encoding 11β-HSD-1 was identified in all the major WAT depots in dogs and also in liver, kidney, and spleen. Quantification by real-time PCR showed that 11β-HSD-1 mRNA was least in perirenal and falciform depots and greatest in subcutaneous, omental, and gonadal depots. Greater expression was seen in the omental depot in female than in male dogs (P = 0.05). Gene expression for 11β-HSD-1 was also seen in adipocytes, from both subcutaneous and visceral depots, differentiated in culture; expression was evident throughout differentiation but was generally greatest in preadipocytes and during early differentiation, declining as cells progressed to maturity. The inflammatory mediators lipopolysaccharide and tumor necrosis factor α had a main stimulatory effect on 11β-HSD-1 gene expression in canine subcutaneous adipocytes, but IL-6 had no significant effect. Treatment with dexamethasone resulted in a significant time- and dose-dependent increase in 11β-HSD-1 gene expression, with greatest effects seen at 24 h (2nM: approximately 4-fold; 20nM: approximately 14-fold; P = 0.010 for both). When subcutaneous adipocytes were treated with the peroxisome proliferator activated receptor γ agonist rosiglitazone, similar dose- and time-dependent effects were noted. However, no effects were seen when adipocytes from the gonadal WAT depot were treated with rosiglitazone. The induction of 11β-HSD-1 expression, by the pro-inflammatory cytokine tumor necrosis factor α and by lipopolysaccharide may have implications for the pathogenesis of obesity and its associated diseases in the dog.     

牦牛不同部位前体脂肪细胞分离鉴定及分化关键基因表达研究

旨在建立牦牛皮下和肌内前体脂肪细胞的体外研究模型,并检测两部位前体脂肪细胞分化过程中关键基因表达量差异,为研究牦牛不同部位脂肪沉积的分子机制提供试验材料和理论依据.本研究通过采取5头18～22月龄健康麦洼公牦牛的皮下脂肪组织和背最长肌组织,利用胶原酶消化,分离皮下和肌内前体脂肪细胞,随后根据细胞来源将细胞分为肌内组和皮...     

山羊RPL26基因生物信息学分析及对肌内脂肪细胞分化的影响

旨在克隆山羊RPL26基因序列并对其在山羊各组织中的表达情况和对山羊脂肪细胞分化的调控作用进行探究.本研究以1周岁简州大耳羊公羊作为试验对象(健康生长状态良好,体重约50 kg,n=3),利用RT-PCR等方法克隆RPL26序列,对基因及蛋白质序列进行生物信息学分析;以山羊各组织cDNA为模板,利用qPCR方法构建组织...     

猪甘油三酯水解酶和激素敏感脂酶基因表达规律的研究

利用半定量RT-PCR法分析比较了甘油三酯水解酶(Triacylglycerol hydrolase,TGH)和激素敏感脂酶(Hormone-sensitive lipase,HSL)基因在不同猪种、不同发育阶段及不同部位脂肪组织中转录表达的差异,探讨其在猪脂肪组织的表达规律。结果显示,脂肪型个体TGHmRNA表达丰度显著低于瘦肉型和杂交型个体,成年猪较初生仔猪低,皮下、腹膜和内脏脂肪组织中TGH表达量依次递增;其变化规律与HSL相同。此外,对分离培养的原代前体脂肪细胞通过诱导分化和油红O染色区分分化状态,分析TGHmRNA表达的时序变化,发现TGH在前脂肪细胞中不转录表达,诱导分化后开始表达,且在诱导分化第4天表达量最高,分化第10天表达量下降,达到峰值的时间较HSL早。结果表明,TGH的表达与个体肥胖程度、年龄、脂肪组织部位以及脂肪细胞分化程度相关,同时,在脂肪细胞分化过程中,TGH表达峰值早于HSL,提示TGH在脂肪细胞发育过程中可能较早承担基础脂解作用。     

Development and evaluation of empirical equations to interconvert between twelfth-rib fat and kidney, pelvic, and heart fat respective fat weights and to predict initial conditions of fat deposition models for beef cattle

The Davis growth model (DGM) simulates growth and body composition of beef cattle and predicts development of 4 fat depots. Model development and evaluation require quantitative data on fat weights, but sometimes it is necessary to use carcass data that are more commonly reported. Regression equations were developed based on published data to interconvert between carcass characteristics and kilograms of fat in various depots and to predict the initial conditions for the DGM. Equations include those evaluating the relationship between the following: subcutaneous fat (SUB, kg) and 12th-rib fat thickness (mm); visceral fat (VIS, kg) and KPH (kg); DNA (g) in intermuscular, intramuscular, subcutaneous, and visceral fat depots and empty body weight; and contributions of fat (kg) in intramuscular (INTRA), SUB, and VIS fat depots and total body fat (kg). The intermuscular fat (INTER, kg) contribution was found by difference. The linear regression equations were as follows: SUB vs. 12th-rib fat thickness (n = 75; P < 0.01) with R(2) = 0.88 and SE = 10.00; VIS vs. KPH (kg; n = 78; P < 0.01) with R(2) = 0.95 and SE = 2.82; the DNA (g) equations for INTER, INTRA, SUB, and VIS fat depots vs. empty body weight (n = 6, 5, 6, and 6; P = 0.08, P < 0.01, P < 0.01, and P = 0.05) with R(2) = 0.57, 0.93, 0.93, and 0.66, and SE = 0.03, 0.003, 0.02, and 0.03, respectively; and initial contribution of INTRA, SUB, and VIS fat depots vs. total body fat (n = 23; P < 0.01) for each depot, with R(2) = 0.97, 0.99, and 0.97, and SE = 0.61, 0.93, and 1.41, respectively. All empirical equations except for DNA were challenged with independent data sets (n = 12 and 10 for SUB and VIS equations and n = 9 for the initial INTER, INTRA, SUB, and VIS fat depots). The mean biases were -2.21 (P = 0.12) and 2.11 (P < 0.01) kg for the SUB and VIS equations, respectively, and 0.05 (P = 0.97), -0.37 (P = 0.27), 1.82 (P = 0.08), and -1.50 (P = 0.06) kg for the initial contributions of INTER, INTRA, SUB, and VIS fat depots, respectively. The random components of the mean square error of prediction were 73 and 26% for the SUB and VIS equations, respectively, and similarly were 99, 85, 62, and 61% for the initial contributions of INTER, INTRA, SUB, and VIS fat depots, respectively. Both the SUB and VIS equations predicted accurately within the bounds of experimental error. The equations to predict initial fat contribution (kg) were considered adequate for initializing the fat depot differential equations for the DGM and other beef cattle simulation models.     

Molecular and histological evidence of brown adipose tissue in adult cats

Brown adipose tissue (BAT) can influence glucose, lipid, and energy metabolism in rodents. Active BAT is now known to be present in adult humans, and interventions targeting BAT are being investigated for the treatment of human obesity and disorders of glucose and lipid metabolism. Domestic cats, like humans, are at increasing risk for obesity and diabetes but little is known about the presence and role of BAT in adult cats. The purpose of this study was to determine if brown adipocytes, identifiable by histological features and molecular markers, were present in the fat depots of adult cats. Adipose tissue samples from intrascapular, perirenal, and subcutaneous depots of eleven 8–12 year old cats (6 lean, 5 obese), were analyzed by real-time PCR for brown adipocyte markers uncoupling protein 1 (UCP1) and Type II iodothyronine 5′deiodinase (D2), by histological examination and by immunohistochemistry for UCP1.UCP1 mRNA was detectable in interscapular and subcutaneous depots in all cats, and in the perirenal depot in 10/11 cats. D2 mRNA was detectable in all depots from all cats. Multilocular adipocytes were identified in the interscapular depots of 4/11 cats and these were positive for UCP1 immunoreactivity. The results demonstrate that UCP1-expressing brown adipocytes are present in multiple depots of adult lean and long-term obese cats, even at 8–12 years of age. It is possible that dietary components or pharmacological agents that influence brown fat activity could exert a relevant biological effect in cats.     

过表达MAT2A基因促进猪肌内脂肪细胞分化的研究

本研究通过构建腺病毒介导的体外超表达载体，探究腺苷甲硫氨酸转移酶2A（methionine adenosyltransferase 2A，MAT2A）基因在猪肌内脂肪细胞分化中的作用。根据GenBank中猪MAT2A基因mRNA序列（登录号：NM_001167650.1）设计引物，提取猪脂肪组织细胞总RNA并反转录获得cDNA，以此为模板进行PCR扩增并连接到pAdTrack-CMV腺病毒穿梭载体中，对重组质粒pAd-MAT2A进行测序鉴定；pAd-MAT2A载体经PacⅠ限制酶酶切线性化，经质粒大片段回收纯化后转染293A细胞进行病毒包装；采用实时荧光定量PCR检测MAT2A基因表达情况，并提取蛋白进行Western blotting分析，取分化第8天的细胞进行油红O染色。结果表明，穿梭载体pAdTrack-CMV-MAT2A构建成功，并能与骨架载体pAdEasy-1实现同源重组；腺病毒载体pAd-MAT2A转染293A细胞后，病毒滴度达到1E+6 PFU/mL，可满足侵染猪肌内脂肪细胞的需要。实时荧光定量PCR和Western blotting结果显示，MAT2A基因mRNA和蛋白水平均显著上调。油红O染色结果显示，过表达MAT2A基因可促进猪肌内脂肪细胞内脂滴聚积。结果表明，腺病毒介导的MAT2A基因过表达在猪肌内脂肪细胞中呈上调趋势，MAT2A基因可促进脂质积累。     

Adipose tissue, longissimus muscle and anterior pituitary growth and function in clenbuterol-fed heifers

The present study was conducted to determine the effects of feeding clenbuterol on adipose tissue and longissimus muscle growth in heifers. For 50 d, 14 heifers were fed either a sucrose-based, clenbuterol supplement or a placebo in which the clenbuterol had been omitted. The heifers were slaughtered in two groups, based on initial weight. Adipose tissue from several anatomical sites and longissimus muscle (depending on slaughter group) were obtained fresh at slaughter. Changes in carcass characteristics elicited by clenbuterol were similar to those reported by others for steers and sheep. Subcutaneous (sc) and intramuscular (im), but not perirenal, adipocytes were smaller and there were more cells per g tissue in the adipose tissue depots of the clenbuterol-fed heifers. Clenbuterol decreased lipogenic enzyme activities, fatty acid-binding protein activity, basal lipolysis and acetate incorporation into glyceride-fatty acids (P less than .05) in sc adipose tissue, but had no effect (P greater than .05) on lipogenesis or lipolysis in im adipose tissue. Clenbuterol elicited a 20% increase in type II myofiber diameters (P less than .05) but had no effect on type I myofiber diameters. In vitro growth hormone release by perifused anterior pituitaries was not affected significantly by long-term in vivo exposure to clenbuterol. These data indicate that a depression in lipogenesis is the mechanism by which clenbuterol decreases subcutaneous fat accretion in cattle.     

Porcine sirtuin 1 gene clone, expression pattern, and regulation by resveratrol

Sirtuin1 (Sirt1) is a NAD-dependent deacetylase that plays important roles in a variety of biological processes. In the current study, we examined tissue-specific and different expression pattern of porcine Sirt1 and the effect of resveratrol (RES) on expression of Sirt1 in porcine adipocytes. The full-length complementary DNA sequence of porcine Sirt1 was 4,024 bp (GenBank accession no: EU030283), with a 2,226-bp open reading frame encoding a 742-AA protein (a predicted molecular mass of 80.9 kDa; GenBank accession no. ABS29571). Comparison of the deduced AA sequence with the corresponding sequences of human, dog, cattle, and mouse Sirt1 showed 82 to 92% similarity. Furthermore, the porcine Sirt1 was highly expressed in porcine brain, to a lesser degree in spleen and white adipose tissue, and had low but detectable expression in liver. In subcutaneous adipose tissue and omental adipose tissue, expression of the porcine Sirt1 mRNA was greater in adult pigs than in young pigs (P < 0.01). In vitro, exposure of cultured adipocytes to 40 and 80 micro M RES for 24 h increased mRNA levels of porcine Sirt1 by 47.86% (P < 0.01) and 91.04% (P < 0.01), respectively. Accordingly, lipid accumulation and NEFA release were decreased (P < 0.05), respectively. After cultures were treated with RES for 48 h, the mRNA level of porcine Sirt1 was increased by 103.84% (P < 0.01) and 148.79% (P < 0.01), respectively. Lipid accumulation was decreased and NEFA release was increased (P < 0.05), respectively. These results provide information needed for manipulating Sirt1 expression in regulating fat deposition in pigs.     

Comparing mRNA levels of genes encoding leptin,leptin receptor,and lipoprotein lipase between dairy and beef cattle

Body weight and fat mass vary distinctly between German Holstein (dairy cattle) and Charolais (beef cattle). The aim of this study was to determine whether the expression of the obese (Ob) gene and lipoprotein lipase (LPL) gene in fat tissues and expression of the long isoform leptin receptor (Ob-Rb) gene in the hypothalamus were different between these two cattle breeds. Body weight and the area of longissimus muscle cross-section of German Holstein were lower (P<0.001), while body fat content, as well as the omental and perirenal fat mass were higher (P<0.001), compared to Charolais. Plasma insulin and leptin levels between two cattle breeds were determined by radioimmunoassay. Compared to Charolais, plasma insulin concentrations were significantly higher (P<0.01), and plasma leptin levels were tended to be higher (P<0.1) in German Holstein. Ob mRNA levels in subcutaneous and perirenal fat depots, but not in the omental fat depot, were significantly higher (P<0.05) in German Holstein than in Charolais. LPL mRNA expression in the perirenal fat depot of German Holstein was greater in abundance than that of Charolais. No significantly different LPL mRNA levels were found in subcutaneous and omental fat depots, and Ob-Rb mRNA levels in the hypothalamus between these two cattle breeds (P<0.05). Both Ob and LPL expression was greater in perirenal and omental fat depots than in the subcutaneous fat depot (P<0.05). Data indicated that in bovine the Ob and LPL gene expression levels in perirenal fats are an important index that is associated with body fat content, while Ob-Rb in hypothalamus is not.