No abstract

The modifications of the hemodynamic system and hydromineral metabolism are physiological features characterizing a normal gestation. Thus, the ability to expand plasma volume without increasing the level of blood pressure is necessary for the correct perfusion of the placenta. The kidney is essential in this adaptation by reabsorbing avidly sodium and fluid. In this study, we observed that the H,K-ATPase type 2 (HKA2), an ion pump expressed in kidney and colon and already involved in the control of the K+ balance during gestation, is also required for the correct plasma volume expansion and to maintain normal blood pressure. Indeed, compared with WT pregnant mice that exhibit a 1.6-fold increase of their plasma volume, pregnant HKA2-null mice (HKA2KO) only modestly expand their extracellular volume (×1.2). The renal expression of the epithelial Na channel (ENaC) α- and γ-subunits and that of the pendrin are stimulated in gravid WT mice, whereas the Na/Cl− cotransporter (NCC) expression is downregulated. These modifications are all blunted in HKA2KO mice. This impeded renal adaptation to gestation is accompanied by the development of hypotension in the pregnant HKA2KO mice. Altogether, our results showed that the absence of the HKA2 during gestation leads to an “underfilled” situation and has established this transporter as a key player of the renal control of salt and potassium metabolism during gestation.

In industrialized countries, a large part of the population is daily exposed to low K(+) intake, a situation correlated with the development of salt-sensitive hypertension. Among many processes, adaptation to K(+)-restriction involves the stimulation of H,K-ATPase type 2 (HKA2) in the kidney and colon and, in this study, we have investigated whether HKA2 also contributes to the determination of blood pressure (BP). By using wild-type (WT) and HKA2-null mice (HKA2 KO), we showed that after 4 days of K(+) restriction, WT remain normokalemic and normotensive (112 ± 3 mmHg) whereas HKA2 KO mice exhibit hypokalemia and hypotension (104 ± 2 mmHg). The decrease of BP in HKA2 KO is due to the absence of NaCl-cotransporter (NCC) stimulation, leading to renal loss of salt and decreased extracellular volume (by 20 %). These effects are likely related to the renal resistance to vasopressin observed in HKA2 KO that may be explained, in part by the increased production of prostaglandin E2 (PGE2). In WT, the stimulation of NCC induced by K(+)-restriction is responsible for the elevation in BP when salt intake increases, an effect blunted in HKA2-null mice. The presence of an activated HKA2 is therefore required to limit the decrease in plasma [K(+)] but also contributes to the development of salt-sensitive hypertension.

In contrast to monogastric species, renal excretion of inorganic phosphate (Pi) in ruminants is low and this could be attributed to an almost complete tubular Pi reabsorption. However, the functional and regulatory basis for this phenomenon has not yet been clarified. Therefore, it was the aim of the present study to characterize the kinetic parameters of the tubular Pi reabsorption system as affected by P or Ca depletion using preparations of renal cortex brush border membrane vesicles (BBMV) from goats and sheep and to compare the data with respective parameters of porcine preparations. Na-dependent Pi uptake into renal cortex BBMV as a function of Pi concentration showed typical Michaelis-Menten kinetic and respective Scatchard plot analysis of the specific Pi uptake revealed linearity indicating the predominant presence of a single type of Pi transporters in the preparations. Under control conditions Vmax values of Na-dependent Pi uptake into BBMV were highest in goats and sheep and lowest in pigs (1.98, 1.39 and 0.95 nmol x mg(-1) protein x 10s(-1), respectively). Km values were not different between goats and sheep under all feeding conditions and ranged from between 0.34 mmol x l(-1) and 0.55 mmol x l(-1) which was three- to five-times higher than that found in pigs (0.11 mmol x l(-1)). Oligonucleotides derived from rat kidney cortex type IIa Na/Pi cDNA were used for reverse transcriptase-polymerase chain reaction (RT-PCR) in goat, sheep and pig kidney cortex. The products isolated were 768 bp for sheep and pigs and 765 bp for goats, with the respective amino acids sequences, representing a segment of approximately 40% in length of the entire transporter, exhibiting an at least 92% sequence homology between different species. From the results, involvement of type IIa Na/Pi cotransport in tubular Pi reabsorption in small ruminants can be postulated. However, it should not be considered that a potential role of other Pi transport systems be completely be excluded. Interestingly, neither P nor Ca depletion caused significant effects on Na-dependent Pi transport capacities and affinities in goats and sheep. From this, parathyroid-hormone independent regulatory pathways of tubular Pi reabsorption can be assumed.

Idiopathic nephrotic syndrome (INS) is characterized by proteinuria and renal Na retention leading to oedema. This Na retention is usually attributed to epithelial sodium channel (ENaC) activation following plasma aldosterone increase. However, most nephrotic patients show normal aldosterone levels. Using a corticosteroid-clamped rat model of INS (CC-PAN), we showed that the observed electrogenic and amiloride-sensitive Na retention could not be attributed to ENaC. We, then, identified a truncated variant of acid sensing ion channel 2b (ASIC2b) that induced sustained acid-stimulated sodium currents when co-expressed with ASIC2a. Interestingly, CC-PAN nephrotic ASIC2b-null rats did not develop sodium retention.We finally showed that expression of the truncated ASIC2b in kidney was dependent on the presence of albumin in the tubule lumen and activation of ERK in renal cells. Finally, the presence of ASIC2 mRNA was also detected in kidney biopsies from patients with INS but not in any of the patients with other renal diseases. We have, therefore, identified a novel variant of ASIC2b responsible for the renal Na retention in the pathological context of INS. Results Sodium retention originates from collecting ducts of CC-PAN rats but is independent ofthe epithelial sodium channel ENaC. Measurement of the net transepithelial flux of sodium (JNa+) by in vitro microperfusion of isolated cortical collecting ducts (CCD) showed that, in contrast to CCDs from control rats in which no JNa+ is measurable (9), CCDs from both PAN and CC-PAN rats displayed significant and similar JNa+ (figure 1A). CCD from PAN and CC-PAN rats also displayed a lumen negative transepithelial voltage (in mV ± SE; PAN: -14.1 ± 2.2, n=8; CC-PAN: -12.9 ± 1.9, n=5; NS), indicating that sodium reabsorption is an electrogenic process. As previously reported in PAN rats (4), luminal addition of amiloride abolished JNa+ in CCD from CC-PAN rats (figure 1B) as well as the lumen negative transepithelial voltage (in mV ± SE; Control: -9.9 ± 4.7; Amiloride: 5.9 ± 0.8, n=3; p<0.03). All these properties are consistent with an amiloride-sensitive process mediating sodium reabsorption in CCD from CC-PAN rats. However previous studies concluded to the absence of functional ENaC under these conditions (6, 8). Using sodium-depleted (LNa) rats as a model of over expression of ENaC in the CCD (10), we searched for properties differentiating ENaC-mediated sodium transport from that in CC-PAN rats. JNa+ in CCD from LNa rats was markedly reduced following luminal addition of 300µM ZnCl2 whereas it was slightly increased in CC-PAN rats (figure 1C).Luminal acidification (pH ≈ 6.0) abolished JNa+ in LNa rats but had no significant effect in CC-PAN rats (figure 1D). Along with pieces of evidence previously reported (6, 8), these findings demonstrate that sodium retention in CC-PAN rats originates from the ASDN and stems from the activation of an electrogenic, amiloride-sensitive, Zn-and pH-insensitive transport pathway independent of ENaC. ASIC2 is responsible for JNa+ and ...

The screening assay was studied in terms of specificity and practicality and was found to be suitable for use in routine testing of blood donations. However, throughput must be enhanced by automation of the assay, and traceability would be improved if automated systems were used to distribute and identify samples.

A low potassium (K+) intake is a common situation in the population of the Westernized countries where processed food is prevalent in the diet. Here, we show that expression of GDF15, a TGFbeta-related growth factor, is increased in renal tubular segments and gut parts of mice in response to low-K+ diet leading to a systemic elevation of its plasma and urine concentration. In human, under mild dietary K+ restriction, we observed that urine GDF15 excretion is correlated with plasma K+ level. Conversely to WT mice, adaptation to K+ restriction of GDF15-KO mice is not optimal, they do not increase their number of type A intercalated cell, responsible for K+ retention, and have a delayed renal K+ retention, leading to early development of hypokalemia. This renal effect of GDF15 depends on ErBb2 receptor, whose expression is increased in the kidney collecting ducts. We also observe that, in the absence of GDF15, the release of K+ by the muscles is blunted which is compensated by a loss of muscle mass. Thus, in this study, we showed that GDF15 plays a central role in the response to K+ restriction by orchestrating the modification of the cell composition of the collecting duct.

We have recently reported that type A intercalated cells of the collecting duct secrete Na+ by a mechanism coupling the basolateral type 1 Na+-K+-2Cl− cotransporter with apical type 2 H+-K+-ATPase (HKA2) functioning under its Na+/K+ exchange mode. The first aim of the present study was to evaluate whether this secretory pathway is a target of atrial natriuretic peptide (ANP). Despite hyperaldosteronemia, metabolic acidosis is not associated with Na+ retention. The second aim of the present study was to evaluate whether ANP-induced stimulation of Na+ secretion by type A intercalated cells might account for mineralocorticoid escape during metabolic acidosis. In Xenopus oocytes expressing HKA2, cGMP, the second messenger of ANP, increased the membrane expression, activity, and Na+-transporting rate of HKA2. Feeding mice with a NH4Cl-enriched diet increased urinary excretion of aldosterone and induced a transient Na+ retention that reversed within 3 days. At that time, expression of ANP mRNA in the collecting duct and urinary excretion of cGMP were increased. Reversion of Na+ retention was prevented by treatment with an inhibitor of ANP receptors and was absent in HKA2-null mice. In conclusion, paracrine stimulation of HKA2 by ANP is responsible for the escape of the Na+-retaining effect of aldosterone during metabolic acidosis.