Channels In Renal Cells Research Articles

AQP2 can modulate the pattern of Ca2+ transients induced by store-operated Ca2+ entry under TRPV4 activation.

There is increasing evidence indicating that aquaporins (AQPs) exert an influence in cell signaling by the interplay with the TRPV4 Ca2+ channel. Ca2+ release from intracellular stores and plasma membrane hyperpolarization due to opening of Ca2+ -activated potassium channels (KCa) are events that have been proposed to take place downstream of TRPV4 activation. A major mechanism for Ca2+ entry, activated after depletion of intracellular Ca2+ stores and driven by electrochemical forces, is the store-operated Ca2+ entry (SOCE). The consequences of the interplay between TRPV4 and AQPs on SOCE have not been yet investigated. The aim of our study was to test the hypothesis that AQP2 can modulate SOCE by facilitating the interaction of TRPV4 with KCa channels in renal cells. Using fluorescent probe techniques, we studied intracellular Ca2+ concentration and membrane potential in response to activation of TRPV4 in two rat cortical collecting duct cell lines (RCCD1 ), one not expressing AQPs (WT-RCCD1 ) and the other transfected with AQP2 (AQP2-RCCD1 ). We found that AQP2 co-immunoprecipitates with TRPV4 and with the small-conductance potassium channel (SK3). We also showed that AQP2 is crucial for the activation of SK3 by TRPV4, leading to hyperpolarization of the plasma membrane. This seems to be relevant to modulate the magnitude of SOCE and is accompanied by TRPV4 translocation to the plasma membrane only in AQP2 expressing cells. These findings open the perspective to further investigate whether the interplay between different AQPs with TRPV4 and KCa channels can be an important mechanism to modulate SOCE with physiological relevance.

Mechanism underlying hypokalemia induced by trimethyltin chloride: Inhibition of H +/K +-ATPase in renal intercalated cells

Trimethyltin chloride (TMT), a byproduct of plastic stabilizers, has caused 67 poisoning accidents in the world; more than 98% (1814/1849) of the affected patients since 1998 have been in China. As a long-established toxic chemical, TMT severely affects the limbic system and the cerebellum; however, its relationship with hypokalemia, a condition observed in the majority of the cases in the last decade, remains elusive. To understand the mechanism underlying hypokalemia induced by TMT, Sprague–Dawley (SD) rats were administered TMT to determine the relationship between H +/K +-ATPase activity and the blood and urine K + concentration and pH, respectively. H +/K +-ATPase protein and mRNA were observed too. In vitro changes to intracellular pH, K + channels in renal cells were measured. The results showed that TMT increased potassium leakage from the kidney, raised urine pH, and inhibited H +/K +-ATPase activity both in vitro and in vivo. In the tested animals, H +/K +-ATPase activity was positively correlated with the decrease of plasma K + and blood pH but was negatively correlated with the increase of urine K + and urine pH ( P < 0.01), while TMT did not change the expression of H +/K +-ATPase protein and mRNA. TMT decreased intracellular pH and opened K + channels in renal intercalated cells. Our findings suggest TMT can directly inhibit the activity of H +/K +-ATPases in renal intercalated cells, reducing urine K + reabsorption and inducing hypokalemia.

PDZ-Binding and Di-Hydrophobic Motifs Regulate Distribution of Kir4.1 Channels in Renal Cells

It was shown previously that the carboxyl-terminal cytoplasmic portion of Kir4.1 determines the localization of basolateral K+ channel in renal distal tubules, which is composed from the assembly of Kir4.1 and Kir5.1. For clarifying the signals for this localization, specific sequence motifs of Kir4.1 were sought. In HEK293T cells, where Kir4.1 showed linear expression on the cell surface, disruption of the carboxyl-terminal PDZ-binding motif induced mostly clustered distribution but did not reduce whole-cell channel activity. Point mutation analysis revealed that serine377 in this motif was responsible for the surface vicinity expression. Disruption of the di-hydrophobic array of valine333/valine334 induced diffuse cytoplasmic distribution and diminished channel activity. Both valine333 and valine334 contributed to this effect. In contrast to the di-hydrophobic motifs of other membrane proteins that facilitate the sorting, valine333/valine334 supported the cell-surface retention. Because both the PDZ-binding and di-hydrophobic motifs participated in the basolateral expression of both Kir4.1 homomer and Kir5.1/Kir4.1 heteromer in MDCK cells, they are thought to be responsible for the localization of basolateral K+ channel in renal distal tubules.