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首页> 外文期刊>American Journal of Physiology >Smooth muscle myosin isoform expression and LC20 phosphorylation in innate rat airway hyperresponsiveness.
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Smooth muscle myosin isoform expression and LC20 phosphorylation in innate rat airway hyperresponsiveness.

机译：先天大鼠气道高反应性中平滑肌肌球蛋白同工型表达和LC20磷酸化。

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Four smooth muscle myosin heavy chain (SMMHC) isoforms are generated by alternative mRNA splicing of a single gene. Two of these isoforms differ by the presence [(+)insert] or absence [(-)insert] of a 7-amino acid insert in the motor domain. The rate of actin filament propulsion of the (+)insert SMMHC isoform, as measured in the in vitro motility assay, is twofold greater than that of the (-)insert isoform. We hypothesized that a greater expression of the (+)insert SMMHC isoform and greater regulatory light chain (LC(20)) phosphorylation contribute to airway hyperresponsiveness. We measured airway responsiveness to methacholine in Fischer hyperresponsive and Lewis normoresponsive rats and determined SMMHC isoform mRNA and protein expression, as well as essential light chain (LC(17)) isoforms, h-caldesmon, and alpha-actin protein expression in their tracheae. We also measured tracheal muscle strip contractility in response to methacholine and corresponding LC(20) phosphorylation. We found Fischer rats have more (+)insert mRNA (69.4 +/- 2.0%) (mean +/- SE) than Lewis rats (53.0 +/- 2.4%; P < 0.05) and a 44% greater content of (+)insert isoform relative to total myosin protein. No difference was found for LC(17) isoform, h-caldesmon, and alpha-actin expression. The contractility experiments revealed a greater isometric force for Fischer trachealis segments (4.2 +/- 0.8 mN) than Lewis (1.9 +/- 0.4 mN; P < 0.05) and greater LC(20) phosphorylation level in Fischer (55.1 +/- 6.4) than in Lewis (41.4 +/- 6.1; P < 0.05) rats. These results further support the contention that innate airway hyperresponsiveness is a multifactorial disorder in which increased expression of the fast (+)insert SMMHC isoform and greater activation of LC(20) lead to smooth muscle hypercontractility.
机译：通过单个基因的可变mRNA剪接产生四种平滑肌肌球蛋白重链（SMMHC）亚型。这些同工型中的两个因运动域中7个氨基酸插入片段的存在[（+）插入]或不存在[（-）插入]而不同。 （-）插入SMMHC同工型的肌动蛋白丝推进速率，如在体外运动测定中所测量的，是（-）插入同工型的两倍。我们假设（+）插入SMMHC亚型的更大表达和更大的监管轻链（LC（20））磷酸化有助于气道高反应性。我们测量了Fischer高反应性和Lewis正常反应性大鼠中对乙酰甲胆碱的气道反应性，并确定了SMMHC亚型mRNA和蛋白表达，以及气管中必需的轻链（LC（17））亚型，h-caldesmon和α-actin蛋白表达。我们还测量了对乙酰甲胆碱和相应的LC（20）磷酸化反应的气管肌肉条收缩性。我们发现Fischer大鼠比Lewis大鼠（53.0 +/- 2.4％; P <0.05）具有（+）插入mRNA（69.4 +/- 2.0％）（平均+/- SE）和（+）含量高44％插入相对于总肌球蛋白蛋白质的同工型。没有发现LC（17）亚型，h-caldesmon和α-肌动蛋白表达的差异。收缩性实验显示，Fischer气管节段的等距力（4.2 +/- 0.8 mN）比Lewis（1.9 +/- 0.4 mN; P <0.05）大，Fischer的LC（20）磷酸化水平更高（55.1 +/- 6.4） ）高于Lewis（41.4 +/- 6.1; P <0.05）大鼠。这些结果进一步支持以下观点：先天性气道高反应性是一种多因素疾病，其中快速（+）插入的SMMHC同工型的表达增加和LC（20）的更大活化导致平滑肌过度收缩。

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