Francis Richard J B, Chatterjee Bishwanath, Loges Niki T, Zentgraf Hanswalter, Omran Heymut, Lo Cecilia W
Laboratory of Developmental Biology, National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA.
Am J Physiol Lung Cell Mol Physiol. 2009 Jun;296(6):L1067-75. doi: 10.1152/ajplung.00001.2009. Epub 2009 Apr 3.
Mucociliary clearance in the adult trachea is well characterized, but there are limited data in newborns. Cilia-generated flow was quantified across longitudinal sections of mouse trachea from birth through postnatal day (PND) 28 by tracking fluorescent microsphere speed and directionality. The percentage of ciliated tracheal epithelial cells, as determined by immunohistochemistry, was shown to increase linearly between PND 0 and PND 21 (R(2) = 0.94). While directionality measurements detected patches of flow starting at PND 3, uniform flow across the epithelia was not observed until PND 7 at a approximately 35% ciliated cell density. Flow became established at a maximal rate at PND 9 and beyond. A linear correlation was observed between the percentage of ciliated cells versus flow speed (R(2) = 0.495) and directionality (R(2) = 0.975) between PND 0 and PND 9. Cilia beat frequency (CBF) was higher at PND 0 than at all subsequent time points, but cilia beat waveform was not noticeably different. Tracheal epithelia from a mouse model of primary ciliary dyskinesia (PCD) harboring a Mdnah5 mutation showed that ciliated cell density was unaffected, but no cilia-generated flow was detected. Cilia in mutant airways were either immotile or with slow dyssynchronous beat and abnormal ciliary waveform. Overall, our studies showed that the initiation of cilia-generated flow is directly correlated with an increase in epithelial ciliation, with the measurement of directionality being more sensitive than speed for detecting flow. The higher CBF observed in newborn epithelia suggests unique physiology in the newborn trachea, indicating possible clinical relevance to the pathophysiology of respiratory distress seen in newborn PCD patients.
成人气管中的黏液纤毛清除功能已得到充分研究,但关于新生儿的数据有限。通过追踪荧光微球的速度和方向性,对出生至出生后第28天(PND)的小鼠气管纵切面的纤毛产生的流动进行了量化。免疫组织化学测定显示,有纤毛的气管上皮细胞百分比在PND 0至PND 21之间呈线性增加(R² = 0.94)。虽然方向性测量在PND 3时检测到了流动斑块,但直到PND 7,当纤毛细胞密度约为35%时,才观察到上皮细胞的均匀流动。在PND 9及以后,流动以最大速率形成。在PND 0至PND 9之间,观察到有纤毛细胞百分比与流速(R² = 0.495)和方向性(R² = 0.975)之间存在线性相关性。PND 0时的纤毛摆动频率(CBF)高于所有后续时间点,但纤毛摆动波形没有明显差异。携带Mdnah5突变的原发性纤毛运动障碍(PCD)小鼠模型的气管上皮显示,有纤毛细胞密度未受影响,但未检测到纤毛产生的流动。突变气道中的纤毛要么不动,要么摆动缓慢且不同步,并且纤毛波形异常。总体而言,我们的研究表明,纤毛产生的流动的起始与上皮细胞纤毛化的增加直接相关,方向性测量比速度测量对检测流动更敏感。在新生儿上皮中观察到的较高CBF表明新生儿气管具有独特的生理学特性,这表明与新生儿PCD患者中出现的呼吸窘迫的病理生理学可能具有临床相关性。