Department of Cell and Neurobiology, Keck School of Medicine, University of Southern California, Los Angeles, CA 90089-9112, USA.
Curr Eye Res. 2013 Jan;38(1):18-26. doi: 10.3109/02713683.2012.725797. Epub 2012 Sep 25.
To investigate the expressional changes of Na(+)/K(+)-ATPase subunits in the lacrimal glands (LG) of term pregnant rabbits.
LG were obtained from term pregnant rabbits and age-matched female control rabbits for laser capture microdissection (LCM), real time RT-PCR, western blot, and immunofluorescence. The mRNA and proteins of α1, α2, β1, β2, and β3 subunits of Na(+)/K(+)-ATPase were detected and quantified.
Although only the mRNA for β3 from whole LG of pregnant rabbits was significantly different from that of normal controls, many mRNA levels for α1, α2, β1, β2, and β3 from acini and epithelial cells from various duct segments that were collected by LCM were significantly different from those of normal control rabbits. Western blots demonstrated that the expressions of all three β subunits were significantly higher in pregnant rabbits, while both α subunits remained unchanged during pregnancy. Interestingly, immunofluorescence results showed that the distribution patterns of all Na(+)/K(+)-ATPase subunits during pregnancy were similar to those of the control rabbits.
Changes were found in mRNA and protein expressions of Na(+)/K(+)-ATPase subunits in LG from term pregnant rabbits and these changes suggest a role in the pregnancy-related LG secretion changes and dry eye symptoms observed in these animals.
研究钠离子/钾离子-ATP 酶亚基在足月妊娠兔泪腺中的表达变化。
采用激光捕获显微切割(LCM)、实时 RT-PCR、Western blot 和免疫荧光技术,从足月妊娠兔和同龄雌性对照兔的泪腺中获取组织。检测和定量分析钠离子/钾离子-ATP 酶的α1、α2、β1、β2 和β3 亚基的 mRNA 和蛋白。
虽然整个妊娠兔泪腺β3亚基的 mRNA 与正常对照组有显著差异,但从 LCM 收集的各种导管段腺泡和上皮细胞中,α1、α2、β1、β2 和β3 的许多 mRNA 水平与正常对照组有显著差异。Western blot 表明,所有三种β亚基在妊娠兔中的表达均显著升高,而α亚基在妊娠期间保持不变。有趣的是,免疫荧光结果表明,所有钠离子/钾离子-ATP 酶亚基在妊娠期间的分布模式与对照组相似。
在足月妊娠兔的泪腺中发现了钠离子/钾离子-ATP 酶亚基的 mRNA 和蛋白表达变化,这些变化提示它们在与妊娠相关的泪液分泌变化和这些动物中观察到的干眼症症状中发挥作用。