Gestational length affects a change in the transepithelial voltage and the rNKCC2 expression pattern in the ascending thin limb of Henle's loop.

To examine whether the functional and morphologic conversion of the neonatal ascending thin limb (ATL) of Henle's loop is related to gestational length, we evaluated the transepithelial voltages (Vts) of ATLs in perinatal mouse, hamster, rabbit, and rat kidneys. In isolated microperfused tubule preparations, Vts of neonatal ATLs were 23.8 +/- 1.4 in mouse, 25.7 +/- 2.2 in hamster, and 18.2 +/- 1.6 mV in rabbit. The influence of gestational length on the Vts and rat Na-K-Cl cotransporter (rNKCC2) expression pattern was also examined in perinatal rats subjected to a prolonged gestation due to either a daily s.c. injection of 5 mg progesterone or ligation of the extremities of the uterine horn. Vts of d 3 neonates were 2.9 +/- 1.0 (p < 0.0001 versus d 0); Vts of d 23 fetuses subjected to ligation were 4.9 +/- 0.8 (p < 0.005 versus d 0); and Vts of d 23 fetuses given progesterone were 3.4 +/- 1.7 mV (p < 0.001 versus d 0). rNKCC2 expression tended to disappear in the renal papillae of d 23 fetuses. Our data demonstrate that the perinatal conversion of the ATL is a phenomenon commonly observed among rodents; furthermore, it is dependent on the gestational length, but unrelated to the birth process.

Chloride-dependent intracellular pH regulation via extracellular calcium-sensing receptor in the medullary thick ascending limb of the mouse kidney.

The extracellular calcium-sensing receptor (CaSR) located in either luminal or basolateral cell membranes of various types of renal tubules including proximal tubules, Henle's loop and collecting ducts has been thought to play a fundamental role in electrolyte metabolism. To further identify the physiological roles of the CaSR, we examined the effects of Ca(2+) and calcimimetics neomycin (Neo), gentamicin and gadolinium chloride (Gd(3+)) on the intracellular pH (pHi) of in vitro microperfused mouse medullary thick ascending limb (mTAL) cells of Henle's loop, by loading the cells with fluorescent pH indicator 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein and measuring the ratio of fluorescence emission at 530 nm after exciting the dye at 490 and 440 nm. In a steady-state condition in Hepes-buffered solution, the pHi in the mTALs was 7.29 +/- 0.04 (n = 9). A concentration of 200 micromol/l Neo in the basolateral side decreased the pHi after 1 min by -0.13 +/- 0.02 (n = 34, p < 0.0001). The other calcimimetics showed similar effects on pHi, whereas none of these calcimimetics in the lumen affected pHi. Na(+) removal or the inhibition of Na(+) and proton transport with amiloride, bumetanide, or bafilomycin did not eliminate the effect of Neo on pHi. On the other hand, Cl(-) removal clearly eliminated the Neo-induced pHi decrease (-0.06 +/- 0.01 vs -0.00 +/- 0.05 in Cl(-) removal, n = 4, p < 0.003). Thus, we have demonstrated for the first time that the CaSR is involved in the regulation of the pHi in the mTAL and requires Cl(-) to exert its effect.

Phylogenetic, ontogenetic, and pathological aspects of the urine-concentrating mechanism.

The urine-concentrating mechanism is one of the most fundamental functions of avian and mammalian kidneys. This particular function of the kidneys developed as a system to accumulate NaCl in birds and as a system to accumulate NaCl and urea in mammals. Based on phylogenetic evidence, the mammalian urine-concentrating mechanism may have evolved as a modification of the renal medulla's NaCl accumulating system that is observed in birds. This qualitative conversion of the urine-concentrating mechanism in the mammalian inner medulla of the kidneys may occur during the neonatal period. Human kidneys have several suboptimal features caused by the neonatal conversion of the urine-concentrating mechanism. The urine-concentrating mechanism is composed of various functional molecules, including water channels, solute transporters, and vasopressin receptors. Abnormalities in water channels aquaporin (AQP)1 and AQP2, as well as in the vasopressin receptor V2R, are known to cause nephrogenic diabetes insipidus. An analysis of the pathological mechanism involved in nephrogenic diabetes insipidus suggests that molecular chaperones may improve the intracellular trafficking of AQP2 and V2R, and, in the near future, such chaperones may become a new clinical tool for treating nephrogenic diabetes insipidus.