Genetic ablation of Slc4a10 alters the expression pattern of transporters involved in solute movement in the mouse choroid plexus

Mutational changes of one transporter can have deleterious effects on epithelial function leaving the cells with the options of either compensating for the loss of function or dedifferentiating. Previous studies have shown that the choroid plexus epithelium (CPE) from mice lacking the Na+-dependent Cl−/HCO3− exchanger (NCBE) encoded by Slc4a10 leads to retargeting of the Na+/H+ exchanger 1 (NHE1) from the luminal to the basolateral plasma membrane. We hypothesized that disruption of NCBE, the main basolateral Na+ importer in the CPE, would lead to a compensatory increase in the abundance of other important transport proteins in this tissue. Aquaporin-1 (AQP1) abundance was 42.7% lower and Na,K-ATPase 36.4% lower in the CPE of Slc4a10 knockout mice, respectively. The NHE1 binding ezrin cytoskeleton appeared disrupted in Slc4a10 knockout mice, whereas no changes were observed in cellular polarization with respect to claudin-2 and appearance of luminal surface microvilli. The renal proximal tubule constitutes a leaky epithelium with high transport rate similar to CPE. Here, Slc4a10 knockout did not affect Na,K-ATPase or AQP1 expression. CPE from AQP1 knockout mice has a secretory defect similar to Slc4a10 mice. However, neither NCBE nor Na,K-ATPase expression was affected in CPE from AQP1 knockout mice. By contrast, the abundance of Na,K-ATPase and NBCe1 was decreased by 23 and 31.7%, respectively, in AQP1 knockout proximal tubules, while the NHE3 abundance was unchanged. In conclusion, CPE lacking NCBE seems to spare the molecular machinery involved in CSF secretion rather than compensate for the loss of the Na+ loader. Slc4a10 knockout seems to be more deleterious to CPE than AQP1 knockout.

Claudin-2 is selectively expressed in proximal nephron in mouse kidney

First published August 15, 2001; 10.1152/ajprenal.00021.2001.—The proximal nephron possesses a leaky epithelium with unique paracellular permeability properties that underlie its high rate of passive NaCl and water reabsorption, but the molecular basis is unknown. The claudins are a large family of transmembrane proteins that are part of the tight junction complex and likely form structural components of a paracellular pore. To localize claudin-2 in the mouse kidney, we performed in situ hybridization using an isoform-specific riboprobe and immunohistochemistry using a polyclonal antibody directed against a COOH-terminal peptide. Claudin-2 mRNA and protein were found throughout the proximal tubule and in the contiguous early segment of the thin descending limb of long-looped nephrons. The level of expression demonstrated an axial increase from proximal to distal segments. In confocal images, the subcellular localization of claudin-2 protein coincided with that of the tight junction protein ZO-1. Our findings suggest that claudin-2 is a component of the paracellular pathway of the most proximal segments of the nephron and that it may be responsible for their uniquely leaky permeability properties.

Na+-K+-ATPase and Na+/Ca2+exchange activities in gills of hyperregulatingCarcinus maenas

Na+-K+-ATPase and Na+/Ca2+exchange activities were studied in gills of Carcinus maenas in seawater (SW) and after transfer to dilute seawater (DSW). Carcinushyperregulates its hemolymph osmolarity through active uptake of Na+, Cl−, and Ca2+. In DSW total Na+-K+-ATPase activity in posterior gills quadrupled; Na+/Ca2+exchange specific activity was unaffected, and total activity increased 1.67-fold. Short-circuit current ( Isc) in voltage-clamped posterior gill hemilamellae was −181 μA/cm2in SW and −290 μA/cm2in DSW and up to 90% ouabain sensitive; conductivity was similar in SW or DSW (42 and 46 mS/cm2, respectively) and representative of a leaky epithelium. The new steady state of hemolymph osmolarity 24 h after DSW transfer was preceded, already 3 h after transfer, by increased Na+-K+-ATPase but not Na+/Ca2+exchange activity. Western blot analysis indicated that the amount of Na+-K+-ATPase protein had increased 2.1-fold in crabs acclimated 3 wk to DSW; however, 4 h after DSW transfer no difference in the amount of Na+-K+-ATPase protein was observed. After DSW transfer branchial cAMP content decreased. A negative correlation between branchial Na+-K+-ATPase activity and cAMP content points to rapid regulation of Na+-K+-ATPase through cAMP-dependent protein kinase A activity. Ca2+transport may depend on the high-capacity Na+/Ca2+exchanger coupled to the versatile sodium pump.

Endocytosis of Polypeptides in Rabbit Nasal Respiratory Mucosa

The nasal respiratory mucosa of the rabbit has a leaky epithelium actively transporting polypeptides by a specific transcytosis probably involved in sampling antigens. The transfer displays saturation kinetics and is abolished by metabolic inhibitors, actin filamet and microtubule disassemblers, inhibitors of vesicle fusion, and receptor recycling;it accepts polypeptide-covered but not uncovered nanoparticles.

Fluid Absorption in the Kidney Proximal Tubule

Fluid absorption in the proximal tubule can be driven by a small osmotic difference between the luminal and interstitial fluids because this leaky epithelium has a high water permeability. The osmotic difference is produced by solute absorption, which tends to dilute the luminal fluid and concentrate the interstitial fluid. However, important questions remain unanswered regarding the pathway for water flow and the role of hemodynamic and humoral factors.

Renal proximal tubule of flounder. I. Physiological properties

The proximal segment of the winter flounder, Pseudopleuronectes americanus, was investigated. Isolated tubules net secrete fluid, although at low rates, 37 pl X min-1 X mm-1. The dominant ions in secreted fluid are Na and Cl, with [Cl] significantly higher than in the bath. Mg and SO4 concentrations in secreted fluid are more than 10-fold greater than in the bath. The transepithelial voltage (-1.9 mV) and resistance (26 omega X cm2) indicate an electrically leaky epithelium, and transepithelial diffusion potentials suggest the Na selectivity of the paracellular pathway. Transepithelial electrochemical potentials point to active transport of Mg, SO4, and probably also Cl and to transepithelial equilibrium of Na. Failure to observe any secretory transport in tubules perfused in vitro at the commonly used perfusion rates illustrates the necessity of low, preferably subnanoliter, perfusion rates in detecting and studying low-capacity epithelial transport systems by the method of Burg.