Acute regulation of OAT3-mediated estrone sulfate transport in isolated rabbit renal proximal tubules

Mahidol University, Krung Thep, Bangkok, Thailand
American journal of physiology. Renal physiology (Impact Factor: 3.25). 12/2004; 287(5):F1021-9. DOI: 10.1152/ajprenal.00080.2004
Source: PubMed


We investigated the regulation of organic anion transport driven by the organic anion transporter 3 (OAT3), a multispecific OAT localized at the basolateral membrane of the renal proximal tubule. PMA, a PKC activator, inhibited uptake of estrone sulfate (ES), a prototypic substrate for OAT3, in a dose- and time-dependent manner. This inhibition was reduced by 100 nM bisindoylmaleimide I (BIM), a specific PKC inhibitor. The alpha(1)-adrenergic receptor agonist phenylephrine also inhibited ES uptake, and this effect was reduced by BIM. These results suggest that PKC activation downregulates OAT3-mediated organic anion transport. In contrast, epidermal growth factor (EGF) increased ES uptake following activation of MAPK. Exposure to PGE(2) or dibutyryl (db)-cAMP also enhanced ES uptake. Stimulation produced by PGE(2) and db-cAMP was prevented by the PKA inhibitor H-89, indicating that this stimulation required PKA activation. In addition, inhibition of cyclooxygenase 1 (COX1) (but not COX2) inhibited ES uptake. Furthermore, the stimulatory effect of EGF was eliminated by inhibition of either COX1 or PKA. These data suggest that EGF stimulates ES uptake by a process in which MAPK activation results in increased PGE(2) production that, in turn, activates PKA and subsequently stimulates ES uptake. Interestingly, EGF did not induce upregulation immediately following phenylephrine-induced downregulation; and phenylephrine did not induce downregulation immediately after EGF-induced upregulation. These data are the first to show the regulatory response of organic anion transport driven by OAT3 in intact renal proximal tubules.

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    • "Also like the OAT1 results, EGF stimulates ES uptake in isolated rabbit renal proximal tubules. The signalling process for this effect is identical to that described above for OAT1 up-regulation (Soodvilai et al. 2004). In our laboratory, we have recently shown that both insulin and EGF are able to stimulate ES transport in rat renal cortical slices through the activation of PKCζ, an atypical PKC that is not activated by phorbol esters, calcium, or DOG (Hirai and Chida 2003; Jenny et al. 2005). "
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