猪苹果酸酶及苹果酸脱氢酶基因在肌肉和脂肪组织中的表达差异
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摘要
本实验采用荧光定量PCR技术(SYBR GreenⅠ法),以ACTB (beta actin,β-肌动蛋白),TBP (TATA box binding protein, TATA框结合蛋白)和TOP2B (topoisomeraseⅡbeta,拓扑异构酶Ⅱ-β)为内参基因,检测了猪苹果酸酶1(malie enzyme1, ME1)苹果酸脱氢酶1和2(malate dehydrogenase, MDH1和MDH2)基因mRNA丰度在7月龄荣昌猪和长白猪不同部位脂肪和肌肉组织中的表达差异,本研究的结果可为进一步探讨ME1、MDH1和MDH2基因与猪肌肉生长与脂肪沉积性状的遗传联系提供基础数据。
结果显示：背膘厚(backfat thickness, BFT)、猪肥胖指数(porcine obesity index,POI)指数和脂肪酸组成差异与荣昌猪脂肪沉积能力较长白猪强,瘦肉率较长白猪低的品种特征相符。猪脂肪和肌肉组织中ME1、MDH1和MDH2基因的nRNA丰度存在明显的性别和品种差异：
(1)品种内相同组织性别间的多重比较表明：雄性长白猪背部皮下脂肪内层、小网膜和肌间脂肪中ME1基因的表达量极显著高于雌性长白猪(P<0.01),雌性长白猪小网膜和肾周脂肪组织中ME1基因表达量极显著高于雄性长白猪(P<0.01)；雌性荣昌猪背部皮下脂肪外层和心脏表面脂肪中ME1基因表达量显著高于雄性荣昌猪(P<0.05),雄性荣昌猪小网膜中ME1基因表达量极显著高于雌性荣昌猪(P<0.01)。雄性长白猪背部皮下脂肪外层中MDH1基因表达量显著高于雌性长白猪(P<0.05),雌性长白猪肌间脂肪中MDH1基因表达量极显著高于雄性长白猪(P<0.01)；雌性荣昌猪背部皮下脂肪外层、肾周脂肪、肌间脂肪和心脏表面脂肪中MDH1基因表达量极显著高于雄性荣昌猪(P<0.01)。雄性长白猪腰大肌中MDH2基因表达量极显著高于雌性长白猪(P<0.01),而在小网膜、肾周脂肪、肌间脂肪、心脏表面脂肪和背最长肌中雌性长白猪MDH2基因的表达量极显著高于雄性长白猪(P<0.01)。其余品种内相同组织性别间ME1、MDH1和MDH2基因表达量差异均未达显著水平(P>0.05)。
(2)相同性别同一组织品种间的多重比较表明：ME1基因在雄性长白猪的小网膜和肌间脂肪的表达量极显著高于雄性荣昌猪(P<0.01)；ME1基因在雌性荣昌猪背部皮下脂肪内层和心脏表面脂肪中的表达量显著高于雌性长白猪(P<0.05),而ME1基因在雌性荣昌猪背部皮下脂肪外层组织中的表达量极显著高于雌性长白猪(P<0.01),ME1基因在雌性长白猪小网膜和肾周脂肪中表达量极显著高于雌性荣昌猪(P<0.01)。MDH1基因在雄性荣昌猪和雌性荣昌猪的肾周脂肪、腹部皮下脂肪和肌间脂肪中的表达量,分别极显著高于雄性长白猪和雌性长白猪(P<0.01)；雌性荣昌猪背部皮下脂肪外层、小网膜和心脏表面脂肪中MDH1基因的表达量极显著高于雌性长白猪(P<0.01)；MDH1基因在雌性荣昌猪背部皮下脂肪内层组织中的表达量显著高于在雌性长白猪(P<0.05)。MDH2基因在雄性荣昌猪背最长肌中的表达量极显著高于雄性长白猪(P<0.01)；雌性荣昌猪小网膜、肾周脂肪、肌间脂肪、心脏表面脂肪和腰大肌中MDH2基因的表达量极显著高于雌性长白猪(P<0.01)。其余相同性别同一组织品种间ME1、MDH1和MDH2基因表达量差异均未达显著水平(P>0.05)。
To investigate the mRNA abundance of porcine malic enzyme 1 (ME1), malate dehydrogenasel and 2 (MDH1 and MDH2) genes across the adipose and muscle tissues at different body site of Landrace (a leaner Western breed) and Rongchang pigs (a fatty indigenous Chinese breed), The real-time fluorescence quantitative PCR was applied. In addition, the relationships betwin on adipose deposition and muscle growth traits and gene expression changes are analysed.
Our results showed that the Rongchang pigs had higher adipocyte deposition capability than that of the Landrace pigs in backfat thickness (BFT), porcine obesity index (POI) and fatty acid composition. The expression changes of porcine ME1, MDH1 and MDH2 mRNA in the adipose and muscle tissues exhibited the specific different patterns by genderes and breeds:
(1) The mRNA abundance of ME1 mRNA in male Landrace pigs was significantly higher than in female Landrace pigs in upper layer of back fat, lesser omentum and intermuscular fat (P<0.01), but the mRNA abundunce of ME1 mRNA in female Landrace pigs was significantly higher than in male Landrace pigs in lesser omentum and leaf fat (P<0.01). The mRNA abundunce of ME1 mRNA in female Rongchang pigs was significantly higher than in male Rongchang pigs in upper layer of back fat and pericardial fat (P<0.05), the mRNA abundunce of ME1 mRNA in male Rongchang pigs was significantly higher than in female Rongchang pigs in lesser omentum (P< 0.01).In upper layer of back fat, the mRNA abufidunce of MDH1 mRNA in male Landrace pigs was significantly higher than in female Landrace pigs (P<0.05), however the mRNA abundunce of MDHl mRNA in female Landrace pigs was significantly higher than in male Landrace pigs in intermuscular fat (P<0.01); the mRNA abundunce of MDH1 mRNA in female Rongchang pigs was significantly higher than in male Rongchang pigs in intermuscular fat, upper layer of back fat, leaf fat and pericardial fat (P< 0.01).In addition psoas major muscle, the mRNA abundunce of MDH2 mRNA in male Landrace pigs was significantly higher than in female Landrace pigs (P< 0.01), the mRNA abundunce of MDH2 mRNA in female Landrace pigs was significantly higher than in male Landrace pigs in intermuscular fat, lesser omentum, leaf fat, pericardial fat and longissimus dorsi muscle (P<0.01).
(2) The mRNA abundunce of ME1 niRNA in male Landrace pigs was significantly higher than in male Rongchang pigs in lesser omentum and intermuscular fat (P<0.01), however, the mRNA abundunce of ME1 mRNA in female Rongchang pigs was significantly higher than in female Landrace pigs in inner layer of back fat and pericardial fat (P<0.05). The mRNA abundunce of MDH1 mRNA in female and male Rongchang pigs was respective significantly higher than in female and male Landrace pigs in abdominal fat, intermuscular fat and pericardial fat (P<0.01). The mRNA abundunce of MDH1 mRNA in female Rongchang pigs was significantly higher than in female Landrace pigs in upper layer of back fat, lesser omentum and pericardial fat (P< 0.01). The mRNA abundunce of MDH2 mRNA in male Rongchang pigs was significantly higher than in male Landrace pigs in longissimus dorsi muscle (P<0.01), the mRNA abundunce of MDH2 mRNA in female Rongchang pigs was significantly higher than in female Landrace pigs in lesser omentum, pericardial fat, intermuscular fat, leaf fat and psoas major muscle (P<0.01).
结果显示：背膘厚(backfat thickness, BFT)、猪肥胖指数(porcine obesity index,POI)指数和脂肪酸组成差异与荣昌猪脂肪沉积能力较长白猪强,瘦肉率较长白猪低的品种特征相符。猪脂肪和肌肉组织中ME1、MDH1和MDH2基因的nRNA丰度存在明显的性别和品种差异：
(1)品种内相同组织性别间的多重比较表明：雄性长白猪背部皮下脂肪内层、小网膜和肌间脂肪中ME1基因的表达量极显著高于雌性长白猪(P<0.01),雌性长白猪小网膜和肾周脂肪组织中ME1基因表达量极显著高于雄性长白猪(P<0.01)；雌性荣昌猪背部皮下脂肪外层和心脏表面脂肪中ME1基因表达量显著高于雄性荣昌猪(P<0.05),雄性荣昌猪小网膜中ME1基因表达量极显著高于雌性荣昌猪(P<0.01)。雄性长白猪背部皮下脂肪外层中MDH1基因表达量显著高于雌性长白猪(P<0.05),雌性长白猪肌间脂肪中MDH1基因表达量极显著高于雄性长白猪(P<0.01)；雌性荣昌猪背部皮下脂肪外层、肾周脂肪、肌间脂肪和心脏表面脂肪中MDH1基因表达量极显著高于雄性荣昌猪(P<0.01)。雄性长白猪腰大肌中MDH2基因表达量极显著高于雌性长白猪(P<0.01),而在小网膜、肾周脂肪、肌间脂肪、心脏表面脂肪和背最长肌中雌性长白猪MDH2基因的表达量极显著高于雄性长白猪(P<0.01)。其余品种内相同组织性别间ME1、MDH1和MDH2基因表达量差异均未达显著水平(P>0.05)。
(2)相同性别同一组织品种间的多重比较表明：ME1基因在雄性长白猪的小网膜和肌间脂肪的表达量极显著高于雄性荣昌猪(P<0.01)；ME1基因在雌性荣昌猪背部皮下脂肪内层和心脏表面脂肪中的表达量显著高于雌性长白猪(P<0.05),而ME1基因在雌性荣昌猪背部皮下脂肪外层组织中的表达量极显著高于雌性长白猪(P<0.01),ME1基因在雌性长白猪小网膜和肾周脂肪中表达量极显著高于雌性荣昌猪(P<0.01)。MDH1基因在雄性荣昌猪和雌性荣昌猪的肾周脂肪、腹部皮下脂肪和肌间脂肪中的表达量,分别极显著高于雄性长白猪和雌性长白猪(P<0.01)；雌性荣昌猪背部皮下脂肪外层、小网膜和心脏表面脂肪中MDH1基因的表达量极显著高于雌性长白猪(P<0.01)；MDH1基因在雌性荣昌猪背部皮下脂肪内层组织中的表达量显著高于在雌性长白猪(P<0.05)。MDH2基因在雄性荣昌猪背最长肌中的表达量极显著高于雄性长白猪(P<0.01)；雌性荣昌猪小网膜、肾周脂肪、肌间脂肪、心脏表面脂肪和腰大肌中MDH2基因的表达量极显著高于雌性长白猪(P<0.01)。其余相同性别同一组织品种间ME1、MDH1和MDH2基因表达量差异均未达显著水平(P>0.05)。
To investigate the mRNA abundance of porcine malic enzyme 1 (ME1), malate dehydrogenasel and 2 (MDH1 and MDH2) genes across the adipose and muscle tissues at different body site of Landrace (a leaner Western breed) and Rongchang pigs (a fatty indigenous Chinese breed), The real-time fluorescence quantitative PCR was applied. In addition, the relationships betwin on adipose deposition and muscle growth traits and gene expression changes are analysed.
Our results showed that the Rongchang pigs had higher adipocyte deposition capability than that of the Landrace pigs in backfat thickness (BFT), porcine obesity index (POI) and fatty acid composition. The expression changes of porcine ME1, MDH1 and MDH2 mRNA in the adipose and muscle tissues exhibited the specific different patterns by genderes and breeds:
(1) The mRNA abundance of ME1 mRNA in male Landrace pigs was significantly higher than in female Landrace pigs in upper layer of back fat, lesser omentum and intermuscular fat (P<0.01), but the mRNA abundunce of ME1 mRNA in female Landrace pigs was significantly higher than in male Landrace pigs in lesser omentum and leaf fat (P<0.01). The mRNA abundunce of ME1 mRNA in female Rongchang pigs was significantly higher than in male Rongchang pigs in upper layer of back fat and pericardial fat (P<0.05), the mRNA abundunce of ME1 mRNA in male Rongchang pigs was significantly higher than in female Rongchang pigs in lesser omentum (P< 0.01).In upper layer of back fat, the mRNA abufidunce of MDH1 mRNA in male Landrace pigs was significantly higher than in female Landrace pigs (P<0.05), however the mRNA abundunce of MDHl mRNA in female Landrace pigs was significantly higher than in male Landrace pigs in intermuscular fat (P<0.01); the mRNA abundunce of MDH1 mRNA in female Rongchang pigs was significantly higher than in male Rongchang pigs in intermuscular fat, upper layer of back fat, leaf fat and pericardial fat (P< 0.01).In addition psoas major muscle, the mRNA abundunce of MDH2 mRNA in male Landrace pigs was significantly higher than in female Landrace pigs (P< 0.01), the mRNA abundunce of MDH2 mRNA in female Landrace pigs was significantly higher than in male Landrace pigs in intermuscular fat, lesser omentum, leaf fat, pericardial fat and longissimus dorsi muscle (P<0.01).
(2) The mRNA abundunce of ME1 niRNA in male Landrace pigs was significantly higher than in male Rongchang pigs in lesser omentum and intermuscular fat (P<0.01), however, the mRNA abundunce of ME1 mRNA in female Rongchang pigs was significantly higher than in female Landrace pigs in inner layer of back fat and pericardial fat (P<0.05). The mRNA abundunce of MDH1 mRNA in female and male Rongchang pigs was respective significantly higher than in female and male Landrace pigs in abdominal fat, intermuscular fat and pericardial fat (P<0.01). The mRNA abundunce of MDH1 mRNA in female Rongchang pigs was significantly higher than in female Landrace pigs in upper layer of back fat, lesser omentum and pericardial fat (P< 0.01). The mRNA abundunce of MDH2 mRNA in male Rongchang pigs was significantly higher than in male Landrace pigs in longissimus dorsi muscle (P<0.01), the mRNA abundunce of MDH2 mRNA in female Rongchang pigs was significantly higher than in female Landrace pigs in lesser omentum, pericardial fat, intermuscular fat, leaf fat and psoas major muscle (P<0.01).
引文
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