Quantification of myosin heavy chain mRNA in somatic and branchial arch muscles using competitive PCR.
Quantification of myosin heavy chain mRNA in somatic and branchial arch muscles using competitive PCR.
复制标题
使用竞争性 PCR 定量体细胞和鳃弓肌中的肌球蛋白重链 mRNA。
DOI：
10.1152/ajpcell.1998.275.1.c68
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
Ryan,AF
中科院分区：
文献类型：
--
作者：
Jung,HH;Lieber,RL;Ryan,AF
本研究的目的是量化的类型和数量的肌球蛋白重链（MHC）mRNA的肌肉内的不同发育起源，以确定是否调控基因表达是可比的。7 MHC亚型进行了分析，在大鼠成年肢体（伸趾长肌，胫骨前肌，比目鱼肌）和非肢体（眼外肌，甲杓肌，隔膜，咬肌）肌肉使用竞争性PCR检测。构建了一种外源性模板，其包括对7种大鼠肌节MHC亚型（β-心脏、2A、2X、2B、眼外、胚胎和新生儿）以及β-肌动蛋白具有特异性的寡核苷酸序列，并将其用作竞争者。只有眼外肌含有所有七种亚型。所有七块肌肉都含有不同百分比的2A型和2X型MHC转录本。比目鱼肌主要含有β-心肌MHC（87.8 ± 2.6%）。眼外肌MHC仅见于眼外肌和甲杓肌，且比例相对较小（分别为7.4 ± 1.5%和4.0 ± 0.7%）。在眼外肌（7.9 ± 0.3%）、甲杓肌（4.4 ± 0.4%）和咬肌（1.0 ± 0.2%）中鉴定出新生儿MHC，在眼外肌（1.2 ± 0.5%）和比目鱼肌（0.6 ± 0.1%）中均鉴定出胚胎MHC。咬肌中MHC mRNA的绝对质量最大（106 pg/0.5 μg RNA），胫骨前肌最小（64 pg/0.5 μg RNA）。这些值表明MHC mRNA占各种骨骼肌中总mRNA库的4%至17%。体细胞和鳃弓肌肉之间的MHC谱的差异表明，肌肉的发育起源可能，至少部分，是负责的MHC表达程序，在成人中实施。注意到β-心脏和2B型MHC转录物在肌肉中的表达之间的反比关系，这表明这两种亚型的表达可能是受免疫调节的。
The purpose of this study was to quantify the type and amount of myosin heavy chain (MHC) mRNA within muscles of different developmental origins to determine whether the regulation of gene expression is comparable. Seven MHC isoforms were analyzed in rat adult limb (extensor digitorum longus, tibialis anterior, and soleus) and nonlimb (extraocular, thyroarytenoid, diaphragm, and masseter) muscles using a competitive PCR assay. An exogenous template that included oligonucleotide sequences specific for seven rat sarcomeric MHC isoforms (β-cardiac, 2A, 2X, 2B, extraocular, embryonic, and neonatal) as well as β-actin was constructed and used as the competitor. Only the extraocular muscle contained all seven isoforms. All seven muscles contained type 2A and type 2X MHC transcripts in varying percentages. As expected, the soleus muscle contained primarily β-cardiac MHC (87.8 ± 2.6%). Extraocular MHC was found only in the extraocular and thyroarytenoid muscles and in relatively small proportions (7.4 ± 1.5% and 4.0 ± 0.7%, respectively). Neonatal MHC was identified in extraocular (7.9 ± 0.3%), thyroarytenoid (4.4 ± 0.4%), and masseter (1.0 ± 0.2%) muscles, and embryonic MHC was identified both in extraocular (1.2 ± 0.5%) and, unexpectedly, in soleus (0.6 ± 0.1%) muscles. Absolute MHC mRNA mass was greatest in the masseter (106 pg/0.5 μg RNA) and least for the tibialis anterior (64 pg/0.5 μg RNA). These values suggest that MHC mRNA represents from 4 to 17% of the total mRNA pool in various skeletal muscles. Differences in MHC profile between somatic and branchial arch muscles suggest that the developmental origin of a muscle may, at least in part, be responsible for the MHC expression program that is implemented in the adult. An inverse relationship between the expression of β-cardiac and type 2B MHC transcripts across muscles was noted, suggesting that the expression of these two isoforms may be reciprocally regulated.