Role of isoprenoids in cytoskeleton integrity and albumin endocytosis by opossum kidney cells.

BACKGROUND: The actin cytoskeleton has been increasingly implicated in endocytic events that are involved in the reabsorption of filtered protein by the proximal tubule. Isoprenylated small G proteins have emerged as key regulators of the actin cytoskeleton. This study examines the role of isoprenoid intermediates in organization of the cytoskeleton, and the effect of modification of the cytoskeleton on albumin endocytosis. METHODS: The effect of lovastatin on cytoskeleton morphology of opossum kidney cells (OK cells) was determined by staining with fluorescent isothiocyanate (FITC)-phalloidin followed by examination using confocal microscopy. Quantitative effects on albumin binding and uptake were determined using a well-established method. RESULTS: Inhibition of isoprenoid synthesis by lovastatin led to morphological disruption of the cytoskeleton with a concentration-dependent decrease in albumin uptake into OK cells. Addition of mevalonate, but not cholesterol, ameliorated the effects of lovastatin on the cytoskeleton and albumin uptake. Selective inhibition of isoprenoid intermediates with a farnesyltransferase inhibitor suggests that geranylgeranylated proteins mediate cytoskeleton organization. CONCLUSION: These results confirm the importance of cytoskeleton integrity in albumin endocytosis in renal proximal tubules. While the present data suggest that synthesis of isoprenoids via the mevalonate pathway is a critical step in maintenance of the cytoskeleton, the role of individual small G proteins in control of the cytoskeleton and other endocytic events is yet to be defined.

Effects of rilmenidine on proximal tubular fluid absorption in rats.

The antihypertensive agent rilmenidine has threefold higher affinity for I(1) imidazoline receptors compared with alpha(2)-adrenoceptors and acts on the central nervous system by reducing sympathetic activity and in the kidney by inhibiting Na(+)/H(+) exchange activity. In the present study, we examined: (i) the effects of luminal and peritubular administration of rilmenidine on fluid absorption in superficial proximal tubules; and (ii) the nature of the receptors involved in mediating the action of this drug in the presence of specific antagonists (efaroxan, idazoxan and 2-methoxy-idazoxan). Studies were performed in anaesthetized Sprague-Dawley rats using shrinking split-drop micropuncture. Luminal administration of rilmenidine (10(-5) and 10(-13) mol/L) inhibited proximal tubular fluid absorption. Peritubular rilmenidine at 10(-12) and 10(-13) mol/L also inhibited fluid uptake, whereas rilmenidine at 10(-11) mol/L had a significant stimulatory action. In the presence of the I(2) > I(1)/alpha(2)-adrenoceptor antagonist idazoxan (10(-5) mol/L), luminal rilmenidine (10(-5) mol/L) stimulated fluid absorption. Stimulation of fluid uptake was also observed when rilmenidine (10(-5) mol/L) and the I(1) imidazoline receptor antagonist efaroxan (10(-5) mol/L) were added together in the luminal fluid. Luminal administration of the selective alpha(2)-adrenoceptor antagonist 2-methoxy-idazoxan (10(-5) mol/L) resulted in significant attenuation of the inhibitory action of luminal rilmenidine (10(-5) mol/L). This indicates that both I(1) imidazoline receptors and alpha(2)-adrenoceptors are involved in the luminal actions of rilmenidine. The effects of luminal and peritubular administration of alpha-methylnoradrenaline (an alpha(2)-adrenoceptor agonist) were compared with those of rilmenidine. Luminal alpha-methylnoradrenaline, at higher concentrations (10(-7) and 10(-5) mol/L), inhibited fluid absorption, as was seen with peritubular rilmenidine, but, in contrast with rilmenidine, no stimulatory action was observed. Peritubular alpha-methylnoradrenaline inhibited fluid uptake at higher concentrations (10(-5) and 10(-7) mol/L), whereas rilmenidine at these concentrations had no effect. The differences in the concentration-dependent responses for rilmenidine and alpha-methylnoradrenaline indicate that both imidazoline receptors and alpha(2)-adrenoceptors are involved in the actions of these compounds on proximal fluid uptake.