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Changes in mRNA expression of nicotinic acetylcholine receptor subunits during embryonic development of mouse masseter muscle

机译:小鼠咬肌肌肉胚胎发育过程中烟碱乙酰胆碱受体亚基mRNA表达的变化

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Nicotinic acetylcholine receptors (nAChRs) switch from the embryonic-type (alpha(2)betagammadelta subunits) to the adult-type (alpha(2)betaepsilondelta subunits), and disappear besides the neuromuscular junctions with the development of trunk and limb skeletal muscles. However, little is known about this process during the embryonic development of masseter muscle. To identify the time course of the nAChR transition from embryonic day (E) 11 to the newborn stage in mouse masseter muscle, we analyzed the expression level of delta, epsilon, and gamma subunit mRNAs by competitive polymerase chain reaction in combination with reverse transcription as well as distribution of delta subunit protein by immunohistochemistry. The nAChR delta subunit mRNA was initially detected at Ell, showed an approximately 25-fold increase (p<0.0001) between El 1 and El 7, and plateaued thereafter until the newborn stage. Immunostaining for delta subunit was observed in the whole portions of masseter myofibers; at E17 and birth, suggesting that the nAChR elimination does not begin even at the newborn stage. The epsilon subunit mRNA initially appeared at E17, and increased in quantity by 144% (p<0.0001) up to the newborn stage. The quantity of gamma subunit mRNA increased by approximately 240% (p<0.0001) between E11 and E17, and then decreased by 22% (p<0.05) from E17 value at the newborn stage. The beginning of the expression of the epsilon subunit mRNA was coincident with the beginning of the decrease in the quantity of the gamma subunit mRNA, suggesting that the nAChR subunit switch begins at E17. [References: 26]
机译:烟碱型乙酰胆碱受体(nAChRs)从胚胎型(alpha(2)betagammadelta亚基)转变为成人型(alpha(2)betaepsilondelta亚基),并在神经肌肉连接处消失,并伴有躯干和四肢骨骼肌的发育。但是,在咬肌的胚胎发育过程中对该过程知之甚少。为了确定nAChR从胚胎第(E)11天到小鼠咬肌中新生阶段过渡的时间过程,我们通过竞争性聚合酶链反应结合逆转录分析了Δ,ε和γ亚基mRNA的表达水平,以及通过免疫组织化学分析δ亚基蛋白的分布。最初在Ell处检测到nAChR delta亚基mRNA,在El 1和El 7之间显示出大约25倍的增加(p <0.0001),此后一直稳定直到新生阶段。在整个咬肌肌纤维中观察到δ亚单位的免疫染色。在E17和出生时,提示nAChR消除甚至在新生儿阶段就没有开始。 ε亚基mRNA最初出现在E17,直至新生阶段数量增加了144%(p <0.0001)。在E11和E17之间,γ亚基mRNA的数量增加了约240%(p <0.0001),然后在新生儿阶段从E17值减少了22%(p <0.05)。 ε亚基mRNA表达的开始与γ亚基mRNA数量减少的开始相吻合,这表明nAChR亚基转换从E17开始。 [参考:26]

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