Differential expression of calcium-activated chloride channels (CLCA) gene family members in the small intestine of cystic fibrosis mouse models
Differential expression of calcium-activated chloride channels (CLCA) gene family members in the small intestine of cystic fibrosis mouse models
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囊性纤维化小鼠模型小肠中钙激活氯离子通道（CLCA）基因家族成员的差异表达
DOI：
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发表时间：
2006
影响因子：
2.3
通讯作者：
A. Gruber
中科院分区：
文献类型：
--
作者：
I. Leverkoehne;Hannah Holle;F. Anton;A. Gruber
钙激活氯离子通道(CLCA)家族的成员被认为是囊性纤维化(CF)表型的调节因子。本研究采用实时定量RT-PCR方法检测了小鼠mCLCA1、mCLCA2、mCLCA3和mCLCA4在Cf小鼠(cftrtm1Cam、cftrTgH(Neim)1Hgu)和野生型C57BL/6、BALB/c、DBA/2和NMRI小鼠小肠中的表达水平。四种CLCA同源物在不同野生型菌株间的表达水平存在明显差异。CF中mCLCA1和mCLCA4的表达与野生型相似。相反，在所有的CF模型中，mCLCA3mRNA的拷贝数都增加了3倍。连续切片上mCLCA3和PAS反应的免疫组织化学检测发现，mCLCA3表达杯状细胞的数量也有类似的增加，这表明mCLCA3的mRNA拷贝数增加是由于杯状细胞增殖而不是转录调节事件。然而，mCLCA2mRNA拷贝数的增加被认为更有可能是由于转录上调。通过使用组蛋白3磷酸化抗体和激活的caspase-3抗体的免疫组织化学方法确定，mRNA拷贝数的变化与细胞动力学的改变无关。结果提示，mCLCA2和mCLCA3均可作为CF肠道表型的修饰物。
Members of the family of calcium-activated chloride channels (CLCA) have been implicated as modulators of the phenotype in cystic fibrosis (CF). Here, the expression levels of the murine mCLCA1, mCLCA2, mCLCA3 and mCLCA4 were quantified by real-time RT-PCR in the small intestines of CF (cftrtm1Cam, cftrTgH(neoim)1Hgu) and wild type C57BL/6, BALB/c, DBA/2 and NMRI mice. Markedly different expression levels of all four CLCA homologs were observed between the different wild type strains. Expression of mCLCA1 and mCLCA4 was similar in CF versus wild type. In contrast, mCLCA3 mRNA copy numbers were increased up to threefold in all CF models. Immunohistochemical detection of mCLCA3 and PAS reactions on consecutive tissue sections identified a similar increase in mCLCA3 expressing goblet cells, suggesting that elevated mRNA copy numbers of mCLCA3 are due to goblet cell hyperplasia rather than transcriptional regulatory events. Increased mCLCA2 mRNA copy numbers, however, were considered more likely to be due to transcriptional upregulation. Changes in mRNA copy numbers were not associated with altered cell kinetics as determined by immunohistochemistry using antibodies to phospho-histone 3 and activated caspase-3. The results suggest that both mCLCA2 and mCLCA3 may act as modifiers of the intestinal phenotype in CF.
DOI：
10.1152/ajpcell.1999.276.6.c1261
发表时间：
1999-06
期刊：
American journal of physiology. Cell physiology
影响因子：
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作者：
A. Gruber;K. D. Schreur;H. Ji;C. Fuller;B. Pauli
通讯作者：
A. Gruber;K. D. Schreur;H. Ji;C. Fuller;B. Pauli
使用延伸因子 1 α (EF-1 α) 对异质组织样品中 mRNA 可及性进行原位评估。
影响因子：
2.3
作者：
Gruber,AD;Levine,RA
通讯作者：
Levine,RA
影响因子：
4.4
作者：
Gruber, AD;Elble, RC;Pauli, BU
通讯作者：
Pauli, BU