Molecular Characterization and Expression Pattern of Na+-K+-2Cl- Cotransporter 2 (NKCC2) in the Intestine of Starry Flounder Platichthys stellatus after Bacterial Challenge
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We identified Na+-K+-2Cl- cotransporter 2 (NKCC2) cDNA isoform from starry flounder Platichthys stellate. The NKCC2 cDNA encoded a polypeptide of 1,043 amino acids representing 12 putative transmembrane domains based on the bioinformatic topology prediction. In addition, starry flounder NKCC2 possessed highly conserved residues within the transmembrane domain 4, known as an essential site for its functionality. End-point RT-PCR analysis revealed that NKCC2 transcript was only found in anterior and posterior intestines and rectum, with the moderate expression levels. In response to Streptococcus parauberis challenge, the NKCC2 mRNA level in the rectum, but not in other segments, was significantly induced at 3 day post challenge, indicating that excess salt may be transported into the rectum. Taken together, our data indicate that S. parauberis infection would tip the intestinal fluid balance in favor of fluid accumulation, indicating that the bacterial pathogens can interfere with osmotic balance in the intestine as well as normal mucosal immune homeostasis.
Abstract
We identified Na+-K+-2Cl- cotransporter 2 (NKCC2) cDNA isoform from starry flounder Platichthys stellate. The NKCC2 cDNA encoded a polypeptide of 1,043 amino acids representing 12 putative transmembrane domains based on the bioinformatic topology prediction. In addition, starry flounder NKCC2 possessed highly conserved residues within the transmembrane domain 4, known as an essential site for its functionality. End-point RT-PCR analysis revealed that NKCC2 transcript was only found in anterior and posterior intestines and rectum, with the moderate expression levels. In response to Streptococcus parauberis challenge, the NKCC2 mRNA level in the rectum, but not in other segments, was significantly induced at 3 day post challenge, indicating that excess salt may be transported into the rectum. Taken together, our data indicate that S. parauberis infection would tip the intestinal fluid balance in favor of fluid accumulation, indicating that the bacterial pathogens can interfere with osmotic balance in the intestine as well as normal mucosal immune homeostasis.
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