Umenishi F, Carter E P, Yang B, Oliver B, Matthay M A, Verkman A S
Department of Medicine, University of California, San Francisco 94143-0521, USA.
Am J Respir Cell Mol Biol. 1996 Nov;15(5):673-9. doi: 10.1165/ajrcmb.15.5.8918374.
Three members of the water channel (aquaporin) family are expressed in adult rat lung: CHIP28 (AQP-1), MIWC (AQP-4), and AQP-5. Because water channels may be important in the clearance of fluid from the newborn lung, the expression of water channels just before and after birth was investigated using the ribonuclease (RNAse) protection assay. RNA was isolated from lungs, brain, and heart of prenatal rats (fetal days F19, F20, and F21) and postnatal rats (days +1, +2, +5, +7, +21, and adult). Transcript expression was measured relative to a beta-actin control by quantitative densitometry. Whereas beta-actin mRNA expression was nearly constant over time, distinct expression patterns were observed for the three water channels. CHIP28 mRNA expression rose slowly from days F19 to +1, then strongly at day +2, and remained elevated over the first week. MIWC mRNA was weakly expressed prenatally, but strongly increased just after birth. AQP-5 mRNA increased slowly and monotonically between days F20 and +7. These patterns contrasted sharply with the developmental expression of CHIP28 in heart, which decreased over time, and MIWC in brain. Immunocytochemistry showed CHIP28 protein expression in capillary endothelia and MIWC in airway epithelia by day +1; quantitative immunoblot analysis showed increased CHIP28 protein expression over time. These findings are consistent with a role of lung water channels in perinatal fluid clearance; however, proof of physiologic significance will require functional measurements of air space-capillary water permeability.
水通道(水孔蛋白)家族的三个成员在成年大鼠肺中表达:CHIP28(AQP-1)、MIWC(AQP-4)和AQP-5。由于水通道可能在新生儿肺清除液体过程中起重要作用,因此使用核糖核酸酶(RNAse)保护试验研究了出生前后水通道的表达情况。从产前大鼠(胎龄F19、F20和F21)以及产后大鼠(出生后第1、2、5、7、21天和成年)的肺、脑和心脏中分离RNA。通过定量光密度法相对于β-肌动蛋白对照测量转录本表达。虽然β-肌动蛋白mRNA表达随时间几乎保持恒定,但观察到三种水通道有明显不同的表达模式。CHIP28 mRNA表达从F19天到出生后第1天缓慢上升,然后在第2天强烈上升,并在第一周保持升高。MIWC mRNA在产前表达较弱,但出生后立即强烈增加。AQP-5 mRNA在F20天到出生后第7天之间缓慢且单调增加。这些模式与CHIP28在心脏中的发育表达(随时间下降)以及MIWC在脑中的发育表达形成鲜明对比。免疫细胞化学显示,到出生后第1天,CHIP28蛋白在毛细血管内皮中表达,MIWC在气道上皮中表达;定量免疫印迹分析显示CHIP28蛋白表达随时间增加。这些发现与肺水通道在围产期液体清除中的作用一致;然而,生理意义的证据将需要对气腔-毛细血管水通透性进行功能测量。