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Articles in PresS, published online ahead of print June 4, 2002
Am J Physiol Renal Physiol, 10.1152/ajprenal.00107.2002
Submitted on March 19, 2002
Accepted on June 3, 2002
1 Department of Medicine, Laval University, Quebec, Quebec, Canada
2 Department of Cellular and Molecular Physiology, Yale University, New Haven, CT, USA
* To whom correspondence should be addressed. E-mail: paul.isenring{at}crhdq.ulaval.ca.
The Na-K-Cl cotransporter isoform 1 (NKCC1) has been isolated from several species including Squalus acanthias (sa). A second kidney-specific isoform (NKCC2) has been cloned mainly from higher vertebrates. Here, we have isolated the saNKCC24 and found that it is produced in at least 4 spliced variants (saNKCC2A, F, AF, and AFno8)4 of ~1090 residues. Expression of these transcripts in Xenopus laevis oocytes revealed that only "A" and "F" are functional, and that they are more active after incubation in low Cl or hyperosmolar media. Interestingly, rates of activation following exposure to these media were exceptionally rapid, demonstrating for the first time that the NKCC2 itself represents an important site of regulation by Cl and that extracellular domains are involved. Another remarkable finding in this study was the failure to identify NKCC2B, a variant found in the kidney of higher vertebrates and expressed specifically in macula densa cells. This result, in conjunction with the fact that the shark kidney lacks well-developed juxtaglomerular apparati, suggests that the "B" exon evolved as a result of selective pressure presumably by gene duplication of the "A" exon), and that a restricted relationship exists between NKCC2B and macula densa.
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