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Developmental regulation of cardiac MAP4 protein expression.

机译：心脏MAP4蛋白表达的发育调控。

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It has been shown that the level of expression of microtubule-associated protein 4 (MAP4) mRNAs changes throughout neonatal heart development [Chapin SJ, et al. 1995. Biochemistry 34:2289]. In the present study, both immunofluorescence and western blotting methods were used to monitor MAP4 protein expression levels in the developing heart. By both methods, it was shown that the levels of total MAP4 protein were maximal during the first postnatal week, and then declined progressively to adulthood. In addition, four major electrophoretic species that reacted with MAP4-specific antibodies (called bands 1-4) were observed in all heart tissue samples. Three of the four bands decreased in abundance throughout postnatal development, but at different rates. The fourth band remained relatively constant in abundance with increasing postnatal age. To determine if phosphorylation events might contribute to this heterogeneity, western blotting experiments using phospho-specific antibodies and phosphatase digestion of extract samples were performed. No phosphorylation-specific antibody staining was observed and no significant changes were demonstrated in the bands after phosphatase treatment, implying that the observed complexity was due mainly to alternative start site or differential isoform expression. Finally, it was discovered that cardiomyocyte MAP4 associated with drug- and cold-stable microtubules in early neonatal myocytes. Thus, the complex regulation of MAP4 protein expression may play a key role in the functional differentiation of myocyte microtubules during heart development.
机译：研究表明，在整个新生儿心脏发育过程中，微管相关蛋白4（MAP4）mRNA的表达水平都发生了变化[Chapin SJ，et al。 1995.Biochemistry 34：2289]。在本研究中，免疫荧光和蛋白质印迹方法均用于监测发育中的心脏中MAP4蛋白的表达水平。两种方法均显示，在产后的第一周中，总MAP4蛋白水平最高，然后逐渐下降到成年。此外，在所有心脏组织样品中均观察到与MAP4特异性抗体发生反应的四种主要电泳物质（称为条带1-4）。在整个产后发育过程中，四个谱带中的三个谱带的丰度下降，但速率不同。随着出生年龄的增加，第四频段的相对丰度保持相对恒定。为了确定磷酸化事件是否可能导致这种异质性，进行了使用磷酸特异性抗体和提取物样品的磷酸酶消化的蛋白质印迹实验。没有观察到磷酸化特异性抗体染色，在磷酸酶处理后的条带中没有发现明显变化，这表明观察到的复杂性主要是由于替代的起始位点或差异同工型表达。最后，我们发现，心肌细胞MAP4与早期新生儿心肌细胞中的药物稳定和冷稳定微管有关。因此，在心脏发育过程中，MAP4蛋白表达的复杂调节可能在心肌微管的功能分化中起关键作用。

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