The effect of hyperammonemia on myostatin and myogenic regulatory factor gene expression in broiler embryos.
The effect of hyperammonemia on myostatin and myogenic regulatory factor gene expression in broiler embryos.
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DOI：
10.1017/s1751731115000117
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发表时间：
2015-06
期刊：
影响因子：
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通讯作者：
Mozdziak PE
中科院分区：
文献类型：
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作者：
Stern RA;Ashwell CM;Dasarathy S;Mozdziak PE
肌生成由四种肌生成调节因子促进，并被肌生成抑制素显著抑制。本研究的目的是检查胚胎基因调控的肌肉生长抑制素/生肌调节因子，和随后的操作蛋白质合成，在肉鸡胚胎诱导高氨血症。从胚胎第15天（艾德）或第17天开始，在48小时内向肉鸡蛋注射乙酸铵溶液4次。当与假注射对照相比时，对于ED 17和ED 19收集的样品，注射乙酸铵的胚胎中的血清氨浓度显著更高（P < 0.05）。从实验和对照胚胎的胸大肌中提取的mRNA的表达，采用实时定量PCR测量肌生长抑制素、生肌调节因子生肌因子5、生肌决定因子1、生肌素、生肌调节因子4和配对框7。在ED 17和ED 19收集的样品中，肌生长抑制素表达显著降低（P < 0.01）伴随着较高的血清氨浓度。在给予乙酸铵的ED 17收集的样品中，肌生成因子5表达更高（P < 0.05）。在ED 17和ED 19采集的样品中，在乙酸铵注射的胚胎中，生肌调节因子4较低（P ≤ 0.05）。对于任一样本采集年龄，在治疗组之间未观察到生肌决定因子1、生肌蛋白或配对框7表达的显著差异。此外，从处理和对照胚胎中采集的组织学样品的BrdU染色没有显著差异。Westernblot分析显示，与对照组相比，肌生成抑制素的表达水平明显降低（P < 0.05）。总的来说，似乎可以通过高氨血症抑制肌生长抑制素的表达，这有望对胚胎肌发生和出生后肌肉生长产生积极影响。
Myogenesis is facilitated by four myogenic regulatory factors and is significantly inhibited by myostatin. The objective of the current study was to examine embryonic gene regulation of myostatin/myogenic regulatory factors, and subsequent manipulations of protein synthesis, in broiler embryos under induced hyperammonemia. Broiler eggs were injected with ammonium acetate solution four times over 48 hours beginning on either embryonic day (ED) 15 or 17. Serum ammonia concentration was significantly higher (P < 0.05) in ammonium acetate injected embryos for both ED17 and ED19 collected samples when compared to sham-injected controls. Expression of mRNA, extracted from pectoralis major of experimental and control embryos, was measured using real-time quantitative PCR for myostatin, myogenic regulatory factors myogenic factor 5, myogenic determination factor 1, myogenin, myogenic regulatory factor 4, and paired box 7. A significantly lower (P < 0.01) myostatin expression was accompanied by a higher serum ammonia concentration in both ED17 and ED19 collected samples. Myogenic factor 5 expression was higher (P < 0.05) in ED17 collected samples administered ammonium acetate. In both ED17 and ED19 collected samples, myogenic regulatory factor 4 was lower (P ≤ 0.05) in ammonium acetate injected embryos. No significant difference was seen in myogenic determination factor 1, myogenin, or paired box 7 expression between treatment groups for either age of sample collection. Additionally, there was no significant difference in BrdU staining of histological samples taken from treated and control embryos. Myostatin protein levels were evaluated by Western blot analysis, and also showed lower myostatin expression (P < 0.05). Overall, it appears possible to inhibit myostatin expression through hyperammonemia, which is expected to have a positive effect on embryonic myogenesis and postnatal muscle growth.