Protonic conductance across phospholipid bilayer membranes induced by uncoupling agents for oxidative phosphorylation.
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Biomedical subjects
Publications and source records attributed to U Hopfer.
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OBJECTIVE: We sought to determine what type of regulation of transepithelial transport in leaky epithelium can occur at the level of paracellular permeability. METHODS: The epithelial permeability to the polar acid pyranine (Ppyr) and the transepithelial electrical resistance (R) were determined in cultured human cervical epithelium. RESULTS: Extracellular adenosine triphosphate (ATP) acutely and reversibly decreased the paracellular permeability, as evidenced by an increase of R from 10 +/- 3 to 16 +/- 2 omega/cm2 and a decrease in Ppyr from 13 +/- 3 to 9 +/- 3 x 10(-6) cm/sec and (P < .01 for both). The ATP effect was dose related (average median effective concentration 2 +/- 1 microM), saturable at 50 microM, and desensitized with repeated administration; it was mimicked by uridine triphosphate and ATP-gamma-S but not by adenosine monophosphate, adenine, adenosine, or adenosine diphosphate. The ATP effect on resistance remained intact even when the intercellular resistance was decreased with a basal-to-apical pressure gradient, but was abolished by lowering extracellular calcium. CONCLUSIONS: These results indicate the following: 1) Paracellular permeability in the cervical epithelial cell line is regulated by a nucleotide receptor; and 2) the tight junctions are effectors of nucleotide-receptor stimulation. We suggest that extracellular ATP may regulate mucus production in the cervix in vivo by acting on a surface receptor and by increasing the resistance of the tight junctions.
Angiotensin II (AngII) is a potent regulator of electrolyte transport with biphasic effects on salt and HCO3-resorption in proximal tubule epithelia (PCT). In cultured PCT cells, pM to nM AngII activates a GTP-binding protein to inhibit cAMP formation and thus releases inhibition of apical Na/H exchange. Phospholipase A2 is activated by nM to microM AngII releasing arachidonate which is metabolized by a novel P450 epoxygenase to form 5,6-epoxy-eicosatrienoic acid (5,6-EET). 5,6-EET and nM apical AngII cause dihydropyridine-sensitive Ca2+ influx from the extracellular space, inhibition of apical-to-basolateral Na flux, and decrease in epithelial monolayer short circuit current. 5,6-EET also inhibits Na/K-ATPase by 50%. This P450 epoxygenase is physiologically important in the AngII-signaling system because the P450 inhibitor ketoconazole blocks AngII effects while potentiating exogenous 5,6-EET effects. Finally, these AngII-mediated signaling systems are polarized in the PCT with pM basolateral AngII inhibiting adenylate cyclase and nM apical AngII activating PLA2 and subsequent generation of 5,6-EET.
OBJECTIVE: The purpose of the present study was to compare the permeability and regulation of paracellular transport in human cervical cells with those in epithelial cells of other organs. METHODS: Cervical cells (ECE16-1, Caski, and HT3) were grown on filters, and transepithelial electrical conductance (GT) and the permeability to pyranine (PPyr) were determined. RESULTS: Cervical cultures were characterized by high GT (83-125 mS.cm-2) and high PPyr (6.2-18 x 10(-6).sec-1). The GT was not significantly affected by cell density but was increased by 20% by lowering extracellular calcium to 0.45 mmol/L or less. The high values of GT and PPyr and the regulation by extracellular calcium indicate that all three cervical cell lines have "leaky" tight junctional complexes. Addition of extracellular adenosine triphosphate (ATP) at 50 mumol/L to the cervical cultures evoked a biphasic change in GT that was unique to the cervical cells: an initial increase, followed by a sustained decrease by 30% from baseline GT. The decrease of GT was associated with a decrease in PPyr by 17%, indicating that ATP had an effect on the tight junctional/paracellular permeability. The ATP effect was reversible either by washing or by chemical hydrolysis with ATPase. The non-cervical cell lines all responded to extracellular ATP with a transient increase in GT, but not with the pronounced decrease. CONCLUSION: The permeability of the paracellular pathway can be regulated in cervical epithelia by mechanisms that may be different from those in epithelial cells from other organs.