Cooper Deborah S, Lee Hye Jeong, Yang Han Soo, Kippen Joseph, Yun C Chris, Choi Inyeong
Department of Physiology, Emory University School of Medicine, 615 Michael Street, Atlanta, GA 30322, USA.
J Biomed Sci. 2006 Jul;13(4):593-5. doi: 10.1007/s11373-006-9078-6. Epub 2006 Mar 18.
In this study, we examined the tissue-specific expression of two electroneutral Na/HCO(3) cotransporter (NBCn1) variants that differ from each other by the presence of the N-terminal 123 amino acids (cassette II). A rat Northern blot with the probe to nucleotides encoding cassette II detected a 9 kb NBCn1 mRNA strongly in the heart and weakly in skeletal muscles, but absent from most of the tissues including kidney, brain, and pancreas. In the rat heart, PCR with primers flanking cassette II preferentially amplified a DNA fragment that lacked cassette II. However, in the human heart, PCR preferentially amplified a fragment that contained cassette II. This larger PCR product was found virtually in all regions of the human cardiovascular system with strong amplification in the apex, atrium, and atrioventricular nodes. These findings indicate that the variant containing cassette II is almost absent in tissues including brain, kidney, and pancreas, where NBCn1 has been extensively examined.
在本研究中,我们检测了两种电中性钠/碳酸氢根共转运体(NBCn1)变体的组织特异性表达,这两种变体因N端123个氨基酸(盒式结构II)的存在与否而有所不同。用编码盒式结构II的核苷酸探针进行的大鼠Northern印迹分析在心脏中强烈检测到9 kb的NBCn1 mRNA,在骨骼肌中检测到较弱信号,但在包括肾脏、大脑和胰腺在内的大多数组织中未检测到。在大鼠心脏中,用位于盒式结构II两侧的引物进行PCR,优先扩增出一个缺少盒式结构II的DNA片段。然而,在人类心脏中,PCR优先扩增出一个包含盒式结构II的片段。这种较大的PCR产物几乎在人类心血管系统的所有区域都能发现,在心尖、心房和房室结中扩增强烈。这些发现表明,在包括大脑、肾脏和胰腺在内的组织中,几乎不存在含有盒式结构II的变体,而在这些组织中,NBCn1已被广泛研究。