Electroneutral cation-Cl− cotransporters NKCC2β and NCCβ expressed in the intestinal tract of Japanese eel Anguilla japonica

Abstract In the present study, we aimed to elucidate the mechanisms of intestinal Na + and Cl − absorption in Japanese eel, focusing on electroneutral cation-Cl − cotransporters, NKCC2β and NCCβ, expressed in the intestinal tract. First, we cloned cDNAs encoding NKCC2β and NCCβ from the intestinal tract of Japanese eel. In both freshwater- and seawater-acclimated eels, quantitative PCR analysis showed that NKCC2β was predominantly expressed in the anterior and posterior intestines, and that NCCβ expression was specifically high in the rectum. According to immunohistochemistry with anti-eel NKCC2β (reacting with NKCC2β but not with NCCβ) and T4 antibody (reacting with both NKCC2β and NCCβ), NKCC2β was localized in the apical surface of the epithelial cells in the anterior and posterior intestines, whereas NCCβ was likely to be distributed to that in the rectum. Furthermore, a specific NCC inhibitor, hydrochlorothiazide, inhibited of Na + and Cl − absorption, as well as water absorption, in the rectal sac preparations from seawater eel, indicating the involvement of NCCβ in ion absorption in the rectum. Our findings indicate that NKCC2β expressed in the anterior and posterior intestines and NCCβ in the rectum are importantly involved in ion absorption to reduce osmolality of ingested seawater prior to water absorption in seawater-acclimated eel.