Abstract
Expression of pSLC4A4 variant NBCe1B (pancreatic form) is over 30 times larger than that of variant NBCe1A (kidney form) in primary cultures of porcine vas deferens epithelial cells or the PVD9902 cell line. PVD9902 cells constitutively expressing shRNA targeted to a specific 21-bp segment of the common coding sequence shared by porcine SLC4A4 NBCe1A and e1B, revealed, by means of absolute quantitative RT-PCR, a 5.65-fold decrease in the levels of NBCe1B and a 2.55-fold decrease of NBCe1A expression when compared to non-transfected PVD9902 cells, as a clear indication of RNA interference – this cell population is here denominated as pSLC4A4-knockdown (pSLC4A4-kd). HCO3−-dependent ion transport at the basolateral membrane of apical-nystatin permeabilized cells revealed that anion transport in non-transfected cells is 50% larger than in the pSLC4A4-kd cells. Reduction of the HCO3−-dependent anion transport in the apical-nystatin permeabilized model by DNDS is 54% in control cells and 27% in the pSLC4A4-kd. These results indicate that primary and immortalized vas deferens epithelia secreted bicarbonate by an SLC4A4-dependent mechanism that likely influences male fertility. (CFF SCHULT99 and NIH RR-17686 support)
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