Liu P, Hu Y, Grossmann R, Zhao R
Key Laboratory of Animal Physiology and Biochemistry, Nanjing Agricultural University, Nanjing, China Key Laboratory of Poultry Heredity and Breeding, Institute of Poultry Science of Jiangsu Province, Yangzhou, Jiangsu, China Department of Functional Genomics and Bioregulation, Institute of Animal Genetics, FLI, Mariensee, Neustadt, Germany.
J Anim Physiol Anim Nutr (Berl). 2013 Oct;97(5):887-95. doi: 10.1111/j.1439-0396.2012.01334.x. Epub 2012 Aug 1.
To evaluate the effect of maternal leptin on muscle growth, we injected 0 μg (control, CON), 0.5 μg (low leptin dose, LL) or 5.0 μg (high leptin dose, HL) of recombinant murine leptin dissolved in 100 μl of PBS into the albumen of broiler eggs prior to incubation. The newly hatched chicks were all raised under the same conditions until 21 days of age (D21), when body weight was measured and samples of gastrocnemius muscle were collected and weighed. Myosin ATPase staining was applied to identify myofibre types and measure the cross-sectional area (CSA) of myofibres. Real-time PCR was performed to quantify leptin receptor (LEPR), insulin-like growth factor 1 (IGF-1), IGF-1 receptor (IGF-1R), growth hormone receptor (GHR) and myostatin (MSTN) mRNA expression in the gastrocnemius muscle. The activity of calpains (CAPNs) in the gastrocnemius muscle was measured using a quantitative fluorescence detection kit. Male chickens treated with both high and low doses of leptin had significantly higher (p < 0.05) body weight on D21. The high leptin significantly increased the CSA (p < 0.05) of gastrocnemius muscle in male chickens, which coincided with a 93% increase (p < 0.05) in IGF-1 mRNA expression. Likewise, the LL dose increased the weight of gastrocnemius muscle in male chickens (p < 0.05), which was accompanied by a 41% down-regulation (p < 0.05) of MSTN mRNA expression and a decreased activity of CAPNs. However, all these changes were not observed in female chickens. The proportion of myofibre types did not altered. No significant change was detected for LEPR and GHR mRNA expression. These results indicate that in ovo leptin treatment affects skeletal muscle growth in chickens in a dose-dependent and sex-specific manner. The altered expression of IGF-1, MSTN mRNA and activity of CAPNs in skeletal muscle may be responsible for such effects.
为评估母体瘦素对肌肉生长的影响,我们在孵化前将溶解于100 μl PBS中的0 μg(对照,CON)、0.5 μg(低瘦素剂量,LL)或5.0 μg(高瘦素剂量,HL)重组小鼠瘦素注入肉鸡卵的蛋白中。新孵出的雏鸡均在相同条件下饲养至21日龄(D21),此时测量体重,并采集腓肠肌样本称重。应用肌球蛋白ATPase染色来鉴定肌纤维类型并测量肌纤维的横截面积(CSA)。进行实时PCR以定量腓肠肌中瘦素受体(LEPR)、胰岛素样生长因子1(IGF-1)、IGF-1受体(IGF-1R)、生长激素受体(GHR)和肌肉生长抑制素(MSTN)mRNA的表达。使用定量荧光检测试剂盒测量腓肠肌中钙蛋白酶(CAPNs)的活性。接受高剂量和低剂量瘦素处理的雄性鸡在D21时体重显著更高(p < 0.05)。高剂量瘦素显著增加了雄性鸡腓肠肌的CSA(p < 0.05),这与IGF-1 mRNA表达增加93%(p < 0.05)相一致。同样,LL剂量增加了雄性鸡腓肠肌的重量(p < 0.05),同时伴随着MSTN mRNA表达下调41%(p < 0.05)以及CAPNs活性降低。然而,在雌性鸡中未观察到所有这些变化。肌纤维类型的比例未改变。未检测到LEPR和GHR mRNA表达的显著变化。这些结果表明,卵内瘦素处理以剂量依赖性和性别特异性方式影响鸡的骨骼肌生长。骨骼肌中IGF-1、MSTN mRNA表达的改变以及CAPNs的活性可能是造成这种影响的原因。