Response of the porcine MYH4-promoter and MYH4-expressing myotubes to known anabolic and catabolic agents in vitro.
Response of the porcine MYH4-promoter and MYH4-expressing myotubes to known anabolic and catabolic agents in vitro.
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猪MYH4启动子和MYH4表达肌管对已知合成代谢和分解代谢剂的体外反应。
DOI：
10.1016/j.bbrep.2021.100924
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发表时间：
2021-03
影响因子：
2.7
通讯作者：
Brameld JM
中科院分区：
文献类型：
--
作者：
Brearley MC;Loczenski-Brown DM;Loughna PT;Parr T;Brameld JM
肌球蛋白重链-IIB（MyHC-IIB;由MYH 4或Myh 4编码）表达通常与肌肉肥大性生长相关。与其他大型哺乳动物不同，家猪在mRNA和蛋白质水平上表达MyHC-IIB。利用基于荧光的启动子-报告基因系统检测合成代谢和分解代谢试剂对增加猪MYH 4启动子活性的影响，并确定随后是否诱导细胞肥大。将C2 C12成肌细胞与猪MYH 4启动子驱动的ZsGreen和CMV驱动的DsRed表达质粒共转染。在分化开始时，加入处理（二丁酰环AMP（dbcAMP）、Des（1-3）胰岛素样生长因子-1（IGF-I）、三碘-1-甲状腺原氨酸（T3）和地塞米松（Dex））或适当的载体对照，并将细胞维持长达4天。在分化的第4天，收集总荧光和平均肌管直径的测量值，分别作为MYH 4启动子活性和细胞肥大的指标。猪MYH 4启动子活性增加C2 C12肌细胞分化过程中，与第3天和第4天之间的显着增加。MYH 4-启动子活性进一步增加后四天的dbcAMP治疗和平均肌管直径显着增加dbcAMP。猪MYH 4启动子活性也倾向于通过T3处理而增加，但Des（1-3）IGF-I或Dex处理没有影响，而平均肌管直径通过Des（1-3）IGF-I而不是T3或Dex增加。猪MYH 4启动子活性在体外对dbcAMP、Des（1-3）IGF-I和T3处理有反应，如先前在体内研究中所报告的。然而，我们报告说，增加MYH 4启动子活性并不总是与肌肉细胞肥大。基于荧光的报告系统为研究肌细胞肥大性生长提供了有用的工具。体外猪MYH 4启动子-报告系统，用于测试合成代谢和分解代谢剂。猪MYH 4启动子活性和肌管直径的变化串联测量。MYH 4启动子活性对dbcAMP、Des（1-3）IGF-I和T3有反应，如体内所见。MYH 4启动子活性增加并不总是与细胞肥大相关。
Myosin heavy chain-IIB (MyHC-IIB; encoded by MYH4 or Myh4) expression is often associated with muscle hypertrophic growth. Unlike other large mammals, domestic pig breeds express MyHC-IIB at both the mRNA and protein level. To utilise a fluorescence-based promoter-reporter system to test the influence of anabolic and catabolic agents on increasing porcine MYH4-promoter activity and determine whether cell hypertrophy was subsequently induced. C2C12 myoblasts were co-transfected with porcine MYH4-promoter-driven ZsGreen and CMV-driven DsRed expression plasmids. At the onset of differentiation, treatments (dibutyryl cyclic-AMP (dbcAMP), Des(1–3) Insulin-Like Growth Factor-1 (IGF-I), triiodo-l-thyronine (T3) and dexamethasone (Dex)) or appropriate vehicle controls were added and cells maintained for up to four days. At day 4 of differentiation, measurements were collected for total fluorescence and average myotube diameter, as indicators of MYH4-promoter activity and cell hypertrophy respectively. Porcine MYH4-promoter activity increased during C2C12 myogenic differentiation, with a marked increase between days 3 and 4. MYH4-promoter activity was further increased following four days of dbcAMP treatment and average myotube diameter was significantly increased by dbcAMP. Porcine MYH4-promoter activity also tended to be increased by T3 treatment, but there were no effects of Des(1–3) IGF-I or Dex treatment, whereas average myotube diameter was increased by Des(1–3) IGF-I, but not T3 or Dex. Porcine MYH4-promoter activity responded to dbcAMP, Des(1–3) IGF-I and T3 treatment in vitro as observed previously in reported in vivo studies. However, we report that increased MYH4-promoter activity was not always associated with muscle cell hypertrophy. The fluorescence-based reporter system offers a useful tool to study muscle cell hypertrophic growth. In vitro porcine MYH4-promoter-reporter system to test anabolic & catabolic agents. Changes in porcine MYH4-promoter activity & myotube diameter measured in tandem. MYH4-promoter activity responded to dbcAMP, Des(1–3) IGF-I and T3 as seen in vivo. Increased MYH4-promoter activity was not always associated with cell hypertrophy.
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影响因子：
3.7
作者：
Brown DM;Jones S;Daniel ZCTR;Brearley MC;Lewis JE;Ebling FJP;Parr T;Brameld JM
通讯作者：
Brameld JM
影响因子：
4.6
作者：
Brown DM;Williams H;Ryan KJ;Wilson TL;Daniel ZC;Mareko MH;Emes RD;Harris DW;Jones S;Wattis JA;Dryden IL;Hodgman TC;Brameld JM;Parr T
通讯作者：
Parr T
影响因子：
2.7
作者：
Brown, David M.;Parr, Tim;Brameld, John M.
通讯作者：
Brameld, John M.
影响因子：
3.3
作者：
Gunawan, A. M.;Richert, B. T.;Gerrard, D. E.
通讯作者：
Gerrard, D. E.
影响因子：
3.6
作者：
Harrison, AP;Tivey, DR;Dauncey, MJ
通讯作者：
Dauncey, MJ