bluuebunny/crossref_metadata_2025 huggingface dataset:
PAH-alpha-KG countertransport stimulates PAH uptake and net secretion in isolated snake renal tubules
Authors: Chatsudthipong V. , Dantzler W. H.
Published: December 2017

The possibility that p-aminohippurate (PAH)-alpha-ketoglutarate (alpha-KG) countertransport could stimulate basolateral uptake and net transepithelial secretion of PAH was examined in intact isolated nonperfused and perfused snake (Thamnophis spp.) distal proximal renal tubules. Preloading tubules with alpha-KG (100 microM) for 30 min increased [14C]PAH rate of uptake by nonperfused tubules and rate of net secretion by perfused tubules approximately three- to fivefold without increasing cellular ATP content. During stimulation of net secretion, intracellular [14C]PAH concentration increased to same extent as net secretion. Presence of Li+ (2 mM) or absence of Na+ (inhibitors of Na(+)-dicarboxylate cotransport) in bath during alpha-KG preloading eliminated stimulation of PAH transport. Addition of unlabeled alpha-KG to bathing medium stimulated efflux of [14C]PAH across basolateral membrane of tubules with oil-filled lumina, further supporting the concept of PAH-alpha-KG countertransport. Preloading with succinate (100 microM) also stimulated [14C]PAH uptake by nonperfused tubules and net secretion by perfused tubules, but stimulation was only 1.5- to 2-fold and cellular ATP content increased. Moreover, preloading with methyl succinate, a slowly metabolized derivative of succinate, did not stimulate PAH uptake by nonperfused tubules or secretion by perfused tubules. Thus succinate appears to stimulate PAH transport via metabolism, possibly by conversion to alpha-KG, not by direct countertransport. This study indicates for the first time in intact renal tubules that PAH-alpha-KG countertransport can stimulate net PAH secretion by generating an increased intracellular PAH concentration.
Nitric oxide inhibition in rats improves blood pressure and renal function during hypovolemic shock
Authors: Lieberthal W. , McGarry A. E. , Sheils J. , Valeri C. R.
Published: December 2017

We have examined the systemic and renal hemodynamic effects of nitric oxide (NO) inhibition with NG-monomethyl-L-arginine (L-NMMA) in normotensive rats as well as in rats with hypovolemic shock induced by hemorrhage. L-NMMA increased mean arterial blood pressure (MAP) from 114 +/- 4 to 130 +/- 6 mmHg (P less than 0.05) in the nonhemorrhaged rats and from 61 +/- 3 to 89 +/- 3 mmHg (P less than 0.05) in the hypovolemic animals. The absolute increase in MAP was greater in the hypovolemic (31 +/- 3 mmHg) than in the nonhemorrhaged (15 +/- 2 mmHg) rats (P less than 0.05). An excess of L-arginine reversed the increase in MAP induced by L-NMMA in both groups. In the normotensive rats the increase in blood pressure was associated with an elevation in renal vascular resistance (RVR; from 6.5 +/- 0.7 to 8.2 +/- 0.9 mmHg.ml-1.min-1, P less than 0.05) so that renal plasma flow (RPF) and glomerular filtration rate (GFR) were unchanged. In contrast, in the hypotensive rats, the marked increase in MAP induced by L-NMMA infusion was not associated with a significant increase in RVR. As a result L-NMMA increased both RPF (from 6.0 +/- 0.4 to 7.8 +/- 0.4 ml/min, P less than 0.05) as well as GFR (from 1.7 +/- 0.2 to 2.5 +/- 0.2 ml/min, P less than 0.05). We conclude that NO is produced and modulates peripheral resistance in normotensive rats as well as in rats with hypovolemic shock. In the hypovolemic rats NO inhibition substantially improves RPF and GFR.(ABSTRACT TRUNCATED AT 250 WORDS)
11-Dehydrocorticosterone, a glucocorticoid metabolite, inhibits aldosterone action in toad bladder
Authors: Brem A. S. , Matheson K. L. , Barnes J. L. , Morris D. J.
Published: December 2017

The enzyme 11 beta-hydroxysteroid dehydrogenase (11 beta-OHSD) metabolizes glucocorticoid hormones and diminishes their ability to induce sodium transport. In these studies, we determined the location of this enzyme in toad bladder and assessed the biological role for its 11-dehydro end product. Employing a polyclonal antibody directed toward 11 beta-OHSD and immunofluorescence techniques, we located the enzyme in the epithelial cell layer of the toad bladder. Although corticosterone (10(-7) M) can partially suppress aldosterone (10(-7) M)-stimulated short-circuit current (SCC), a clear excess of corticosterone (10(-6) M) did not inhibit the aldosterone-induced induced (10(-8) M) rise in SCC (n = 6). The 11-dehydro product of corticosterone, 11-dehydrocorticosterone (compound A) added to the serosal bath suppressed aldosterone (10(-8) M) peak SCC (360 min) in a dose-dependent fashion reaching 46 +/- 5% of control values at 10(-5) M (n = 6; P less than 0.001). Compound A (10(-5) M) in the mucosal bath also was capable of partially inhibiting the peak aldosterone rise in SCC to 63 +/- 7% of control values with aldosterone at 10(-8) M (n = 6; P less than 0.01) and to 64 +/- 10% of control values with aldosterone at 10(-7) M (n = 9; P less than 0.01). Compound A alone at 10(-5) M did not have any effect on SCC. Isolated toad bladders were not able to transform compound A (at 10(-8) and 10(-5) M) back to corticosterone. Thus the 11-dehydro end product of 11 beta-OHSD (compound A) may play a biologic role by regulating a component of mineralocorticoid-induced sodium transport.
A closed-loop analysis of the tubuloglomerular feedback mechanism
Authors: Holstein-Rathlou N. H.
Published: December 2017

The tubuloglomerular feedback (TGF) mechanism is of importance in the regulation of glomerular filtration rate (GFR). A second mechanism of potential importance is the change in proximal pressure caused by a change, for example, in the rate of proximal fluid reabsorption. The quantitative contributions of these two mechanisms to the regulation of GFR and the late proximal flow rate are not known. To determine the regulatory efficiency of these two mechanisms, the late proximal flow rate was perturbed by microperfusion with artificial tubular fluid in halothane-anesthetized Sprague-Dawley rats. The resulting changes in late proximal flow rate were measured by pulse injection of rhodamine dextran. Fluorescence was excited by means of a He-Ne laser. Bolus velocity was measured by videomicroscopy. Tubular pressure was measured by the servonulling method. The microperfusion rate was varied from -15 to 20 nl/min in steps of 5 nl/min. The open-loop gain (OLG) was 3.1 (range 1.5-9.9, n = 13) at the unperturbed tubular flow rate, and decreased as the tubular flow rate was either increased or decreased. The proximal pressure increased by 0.21 +/- 0.03 mmHg per unit increase in late proximal flow rate (nl/min). By use of a mathematical model of the glomerulus, it is estimated that under the present experimental conditions the pressure increase contributes 8% (range 3-15%) of the OLG. It is concluded that, for small perturbations around the operating point, TGF accounts for most of the regulation of GFR and the late proximal flow rate, with changes in the proximal pressure of lesser importance. Furthermore, under closed-loop conditions the operating point for the TGF mechanism is at or close to the point of maximal sensitivity.
Developmental maturation of Na(+)-H+ exchange in rat renal tubular brush-border membrane
Authors: Zelikovic I. , Stejskal E. , Lohstroh P. , Budreau A. , Chesney R. W.
Published: December 2017

The developmental maturation of the Na(+)-H+ exchanger present in the proximal tubular luminal membrane of the rat was investigated. An overshoot of 1 mM Na+ uptake was evident in brush-border membrane vesicles derived from the renal cortex of 7- and 21-day-old and adult rats in the presence of an outwardly directed H+ concentration ([H+]) gradient [intravesicular pH (pHi) = 5.5; extravesicular pH (pHo) = 7.5]. Na+ uptake was amiloride sensitive at all ages examined. Significantly higher initial rate (3 s) Na+ uptake and peak accumulation (60 s) in the presence of a [H+] gradient were found in vesicles from 7-day-old rats compared with adult animals. Significantly enhanced initial rate Na+ uptake by neonatal vesicles was also evident under pH-equilibrated conditions (pHi = pHo = 7.5). An age-related decrease in amiloride-sensitive Na+ accumulation by vesicles was found. Kinetic analysis of Na(+)-H+ exchange in voltage-clamped vesicles, in the presence of dimethylamiloride (DMA), and calculating 5-s Na+ uptake values showed a maturational decrease in capacity (decreasing Vmax) coupled with a maturational increase in affinity (decreasing Km) of Na(+)-H+ antiport. These data suggest that an enhanced amiloride-inhibitable Na(+)-H+ exchange activity due to increased capacity of exchange exists in the proximal tubular luminal membrane of the neonatal rat. This increased Na(+)-H+ exchange may potentially contribute to positive Na+ balance in the growing organism and may rapidly dissipate the electrochemical Na+ gradient across the luminal membrane necessary for Na(+)-solute contransport, thereby contributing to glycosuria and aminoaciduria of early life.
Multinucleated rat alveolar macrophages express functional receptors for calcitonin
Authors: Vignery A. , Raymond M. J. , Qian H. Y. , Wang F. , Rosenzweig S. A.
Published: December 2017

The fusion of mononuclear phagocytes occurs spontaneously in vivo and leads to the differentiation of either multinucleated giant cells or osteoclasts in chronic inflammatory sites or in bone, respectively. Although osteoclasts are responsible for resorbing bone, the functional role of giant cells in chronic inflammatory reactions and tumors remains poorly understood. We recently reported that the plasma membrane of multinucleated macrophages is, like that of osteoclasts, enriched in Na-K-adenosinetriphosphatases (ATPases). We also observed that the localization of their Na-K-ATPases is restricted to the nonadherent domain of the plasma membrane of cells both in vivo and in vitro, thus imposing a functional polarity on their organization. By following this observation, we wished to investigate whether these cells also expressed, like osteoclasts, functional receptors for calcitonin (CT). To this end, alveolar macrophages were fused in vitro, and both their structural and functional association with CT was analyzed and compared with those of mononucleated peritoneal and alveolar macrophages. Evidence is presented that multinucleated alveolar macrophages express a high copy number of functional receptors for CT. Our results also indicate that alveolar macrophages, much like peritoneal, express functional receptors for calcitonin gene-related peptide. It is suggested that multinucleated rat alveolar macrophages offer a novel model system to study CT receptors and that calcitonin may control local immune reactions where giant cells differentiate.
Effect of reversible ATP depletion on tight-junction integrity in LLC-PK1 cells
Authors: Canfield P. E. , Geerdes A. M. , Molitoris B. A.
Published: December 2017

To further understand and investigate how ischemia affects the tight junction we have developed a 2-h model of rapidly reversible ATP depletion and cellular injury in confluent LLC-PK1 monolayers. ATP depletion was achieved utilizing substrate-free medium containing 0.1 microM antimycin A (AA). Cellular ATP levels dropped rapidly to less than 5% of control values, but recovery of ATP and cell morphology was possible even after 2 h of exposure to AA. Ruthenium red, an electron-dense marker of tight-junction integrity, was excluded from the tight junctions of control monolayers but penetrated cellular tight junctions during ATP depletion in a duration-dependent manner. Electrical resistance across the monolayers remained unchanged in control monolayers but decreased linearly during ATP depletion to 59% of control values. Transmonolayer movement of [3H]mannitol increased from a control level of 7 to 13.5% during ATP depletion. Recovery of tight-junction integrity was demonstrated by a slowing of [3H]mannitol transfer from the basolateral to the apical medium. The transfer rate in control monolayers was 0.0126%/min. During the initial 120 min of cellular recovery from 2 h of ATP depletion, the transfer rate was 0.0789%/min, but this decreased to 0.0045%/min between 2 and 4 h of recovery. In summary, physiology, biochemical, and morphological evidence indicates that reversible ATP depletion results in rapid opening of cellular tight junctions. After ATP-repletion physiological studies indicate a recovery of tight-junction integrity.
Localization of ral, a small Mr GTP-binding protein, to collecting duct cells in bovine and rat kidney
Authors: Gupta A. , Bastani B. , Chardin P. , Hruska K. A.
Published: December 2017

Plasma membranes from bovine kidney cortex were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to nitrocellulose membranes. Blotting with [alpha-32P]GTP and [35S]GTP gamma S demonstrated specific binding to three and six distinct protein bands, respectively, in the 20,000- to 29,000-Mr range. This indicated the presence of small Mr GTP binding proteins (smg) in bovine kidney cortex. Only one smg with 28,000 Mr was labeled with hydrolysis-resistant GTP photoaffinity probe p3-(4-azidoanilido)-p1-5GTP (AAGTP). The major smg in platelet membranes that binds GTP on nitrocellulose blots has been identified as ral-Mr 29,000. With the use of an antiserum against the ral A gene product, one of the smg with Mr of 29,000 present in bovine renal cortical plasma membranes was identified as ral. Ral was absent from glomerular homogenate, suggesting that it is localized to the tubular segments of the nephron. Ral was detected only in the particulate fraction and not the cytosol. Further subcellular localization of ral was investigated by immunohistochemical staining. Anti-ral antibody immunostained the apical and basolateral membranes of cells in the cortical and medullary collecting ducts in a speckled pattern in the bovine kidney. In the rat kidney, however, uniform linear staining of cortical and medullary collecting ducts predominantly localized to the apical membrane was observed. To date, no function has been assigned to ral. Localization of the ral gene product to the collecting duct suggests a specific functional role for this GTP-binding protein.
Expression of cytokine-like genes JE and KC is increased during renal ischemia
Authors: Safirstein R. , Megyesi J. , Saggi S. J. , Price P. M. , Poon M. , Rollins B. J. , Taubman M. B.
Published: December 2017

Both mitogenic and inflammatory phenomena accompany the renal response to ischemic injury. Previous studies have shown that several nuclear-binding members of the immediate early genes are prominently expressed after renal ischemia and may underlie the mitogenic response to such injury. We now report on the expression of JE and KC, other growth-factor-responsive genes that code for small secreted glycoproteins with cytokine-like properties, which may play a role in inflammation. The expression of the immediate early genes JE and KC was determined in rat kidney tissue at varying time points after release of a 50-min period of bilateral renal hilar clamping. Relative levels of mRNA for JE and KC were analyzed by Northern blot analysis of cortical and outer stripe mRNA. KC mRNA rose rapidly to peak values at 1 h and returned toward low baseline levels by 24 h after release of the hilar clamp. By contrast, JE mRNA reached peak levels later and remained elevated for at least 96 h after ischemia. JE antigen was localized immunocytochemically to the apical regions of the cortical and medullary thick ascending limbs as well as in the lumen of the distal nephron in ischemic kidneys. Cells of the glomerulus and proximal tubules were negative for JE antigen. In contrast to the increase in JE and KC mRNA, steady-state levels of uromodulin (Tamm Horsfall) mRNA, a cytokine binding protein also made by the thick ascending limb, declined to virtually undetectable levels by 24 h after ischemia. Thus the increases in JE and KC are not generalized phenomena.(ABSTRACT TRUNCATED AT 250 WORDS)
Effects of atriopeptin III on isolated rat afferent and efferent arterioles
Authors: Lanese D. M. , Yuan B. H. , Falk S. A. , Conger J. D.
Published: December 2017

The effects of atriopeptin III (AP III) on in vitro prepared afferent (AA) and efferent arterioles (EA) from rat kidneys were tested in a system in which lumen diameter could be measured. AP III (10(-13)-10(-7) M) had no effect on lumen diameter of AA that were not preconstricted. When AA were preconstricted with either angiotensin II (ANG II) or norepinephrine (NE), however, AP III increased lumen diameter in a concentration-dependent manner to the preconstriction baseline value. Maximal vasodilation occurred at 10(-10) M AP III. Unlike AA, EA constricted by 50% to 10(-10) M AP III further constricted EA that were pretreated with ANG II or NE. Dilation in ANG II-preconstricted AA to AP III was not inhibited by indomethacin. Constriction of EA to AP III was not altered by [Sar1-Ala8] ANG II, enalapril, OKY 046, or phentolamine. Results indicate that in isolated renal arterioles AP III dilates preconstricted AA but constricts EA that have either not been pretreated or have been preconstricted with other agonists. The effect of AP III on preconstricted AA does not require vasodilator prostaglandin mediation. The constrictor effect of AP III on EA is not dependent on angiotensin, thromboxane, or alpha-adrenergic mediation.
Suppression of growth hormone release restores phosphaturic response to PTH in immature rats
Authors: Mulroney S. E. , Lumpkin M. D. , Haramati A.
Published: December 2017

Immature rats display a blunted rise in urinary phosphate but not adenosine 3',5'-cyclic monophosphate (cAMP) excretion in response to parathyroid hormone (PTH), perhaps as a consequence of the increased demand for phosphate during growth. Because a major driving force for growth is growth hormone (GH), and in view of the fact that GH has been shown to promote renal phosphate retention in the immature animal, it is possible that GH may attenuate the phosphaturic effect of PTH. The objective of this study was to determine whether suppression of pulsatile GH release, during administration of a synthetic peptide antagonist to GH-releasing factor, i.e., [N-acetyl-Tyr1-D-Arg2]-GRF-(1-29)-NH2 (GRF-AN), alters the renal response to increasing doses of PTH (1.5-15.0 micrograms.100 g-1.h-1) in the acutely thyroparathyroidectomized immature rat. Baseline fractional excretion of phosphate (FEPi), before administration of PTH, was negligible in all groups (less than 0.05%). Infusion of PTH resulted in an attenuated rise in FEPi in immature control rats compared with adult control rats (from 3.8 +/- 1.4% at lowest PTH dose to 16.7 +/- 3.1% at highest dose in immature rats vs. 21.1 +/- 3.5 to 31.9 +/- 4.4% in adult rats, P less than 0.05). In contrast, immature rats treated for 2 days with GRF-AN (100 micrograms/kg, twice daily) displayed an enhanced phosphaturic response (FEPi from 12.0 +/- 4.2 to 42.9 +/- 3.7%, P less than 0.05) compared with immature control rats, which was not different from that observed in control adult rats.(ABSTRACT TRUNCATED AT 250 WORDS)
cAMP stimulates protein kinase C activity in cultured renal LLC-PK1 cells
Authors: Anderson R. J. , Breckon R.
Published: December 2017

Exposure of intact LLC-PK1 cells to the phorbol ester 4-phorbol 12-myristate 13-acetate (PMA) increases basal, arginine vasopressin-stimulated, and forskolin-stimulated adenylate cyclase activity in LLC-PK1 membranes. This observation suggests that protein kinase C can increase adenosine 3',5'-cyclic monophosphate (cAMP) in LLC-PK1 cells. To determine whether cAMP regulates protein kinase C activity in LLC-PK1 cells, intact cells were exposed to either forskolin or to soluble cAMP analogues. Acute (5 and 30 min) exposure to either forskolin or cAMP analogues increases protein kinase C activity as observed by two different methods for measuring protein kinase C. Acute exposure to PMA translocates protein kinase C from a soluble to a particulate cell fraction, whereas acute exposure to cAMP increases both soluble and particulate forms of protein kinase C. Longer exposure (18 h) to PMA results in a loss of protein kinase C activity, whereas 18-h exposure to cAMP results in a further increase in protein kinase C activity. The effect of cAMP but not of PMA to stimulate protein kinase C activity can be attenuated by the pro-R diastereoisomer of adenosine 3',5'-cyclic phosphorothioate, suggesting a protein kinase A-mediated effect. These results suggest the presence of a monodirectional mode of signal transduction system interaction in LLC-PK1 cells in which protein kinase C and protein kinase A can potentiate each other.
Taurine attenuates renal disease in chronic puromycin aminonucleoside nephropathy
Authors: Trachtman H. , Del Pizzo R. , Futterweit S. , Levine D. , Rao P. S. , Valderrama E. , Sturman J. A.
Published: December 2017

Repeated administration of low doses of puromycin aminonucleoside (PAMN) to rats induces a proteinuric renal disease that resembles focal segmental glomerulosclerosis (FSGS). Reactive oxygen molecules may be involved in the progressive course of this nephropathy. Therefore we evaluated whether taurine, an endogenous antioxidant, could limit the extent of renal injury. Sprague-Dawley rats received low-dose injections of PAMN, 2 mg/100 g body wt, over a 12-wk period. Two groups were studied: 1) controls given tap water (n = 23), and 2) an experimental group that drank 1% taurine-supplemented water (n = 22). Taurine-treated nephrotic rats had a reduction in albuminuria, as assessed by the urinary albumin-to-creatinine ratio (26 +/- 4 vs. 44 +/- 4, P less than 0.0001). After 12 wk, creatinine clearance was 0.33 +/- 0.03 (experimental) vs. 0.17 +/- 0.03 ml.min-1.100 g body wt-1 (control) (P less than 0.001), and inulin clearance (n = 6 pairs) was 0.26 +/- 0.04 (experimental) vs. 0.13 +/- 0.02 ml.min-1.100 g body wt-1 (control) (P less than 0.025). Administration of taurine reduced the percentage of segmentally sclerosed glomeruli (9.8 +/- 1.7 vs. 16.2 +/- 1.8%, P less than 0.02) and the tubulointerstitial injury score (1.36 +/- 0.19 vs. 2.61 +/- 0.25, P less than 0.0025) in experimental vs. control rats. Taurine treatment normalized the elevated renal cortical malondialdehyde level in rats with PAMN nephropathy (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)
Suppression of blood flow autoregulation plateau during nitric oxide blockade in canine kidney
Authors: Majid D. S. , Navar L. G.
Published: December 2017

We examined the autoregulation of renal blood flow (RBF) and renal function in anesthetized dogs during nitro-L-arginine (NLA)-induced blockade of endothelium-derived nitric oxide (EDNO). Intrarenal infusion of NLA (50 micrograms.kg-1.min-1) increased systemic arterial pressure (AP) and renal vascular resistance (RVR). RBF decreased by 27 +/- 3%, but glomerular filtration rate remained unchanged. There were reductions in urine flow (24 +/- 5%), urinary sodium excretion (42 +/- 10%), and fractional excretion of sodium (40 +/- 11%). The vasodilatory responses to intrarenal injections of ATP (1, 5, 10 microM) were reversed, whereas such responses to doses (10, 50, 100 ng) of acetylcholine (ACh) were attenuated during NLA infusion. Indomethacin (5 mg/kg iv) treatment further reduced but did not completely abolish ACh-induced vasodilation, suggesting that factor(s) other than EDNO and prostaglandins may also mediate ACh-induced vasodilation in the kidney. Although there was a suppression of the plateau of the AP-RBF relationship with a rightward shift in the slope of the linear portion of the curve during EDNO blockade, the normal autoregulatory pattern remained intact. Similar responses were seen in dogs treated with the angiotensin-converting enzyme inhibitor, MK-422. These data indicate that EDNO contributes to the normally low renal vascular tone by influencing an autoregulation-independent component of RVR. However, the basic capability to adjust RVR (autoregulation-responsive component) in response to changes in AP is essentially autonomous from EDNO activity.
Cell polarity and calcium oxalate crystal adherence to cultured collecting duct cells
Authors: Riese R. J. , Mandel N. S. , Wiessner J. H. , Mandel G. S. , Becker C. G. , Kleinman J. G.
Published: December 2017

The relationship between cell membrane polarity and calcium oxalate (CaOx) crystal binding was studied in rat renal inner medullary collecting duct (IMCD) cells in primary culture. Cultures grew as simple monolayers (M) with interspersed cellular aggregates (A), and CaOx bound preferentially to A. An antibody that recognizes an exclusively basolateral epitope in intact IMCD binds to some of the cells in A but not to cells in M. Lysing of intercellular junctions with 3 mM EGTA (monitored by transepithelial resistance, R) resulted in basolateral antibody binding to the previously negative cells in M and a 21-fold increase in CaOx adherence to M over control (P less than 0.01). Enhanced CaOx attachment appeared to lag behind the fall in R by 5–10 min. Crystal attachment returned to control between 30 and 120 min after removal of EGTA and readdition of Ca. These data suggest that loss of epithelial membrane polarity may result in enhanced capacity to bind CaOx. Such loss of cell membrane polarity may occur in IMCD with some forms of epithelial injury and repair and may provide a site of crystal fixation to initiate nephrolithiasis.
Distribution of angiotensin II receptor subtypes in rat and human kidney
Authors: Sechi L. A. , Grady E. F. , Griffin C. A. , Kalinyak J. E. , Schambelan M.
Published: December 2017

Angiotensin II initiates a variety of physiological effects in the kidney by binding to high-affinity receptors on plasma membranes. Recently, two subtypes of angiotensin II receptors have been distinguished on the basis of differences in signal transduction mechanisms, binding affinity to agonists and antagonists, and inhibition of binding by dithiothreitol. To evaluate the density and distribution of these receptor subtypes in the kidney, we performed an in situ autoradiographic study on frozen tissue sections obtained from rat and human kidneys. Sections were incubated with 125I-[Sar1,Ile8]angiotensin II and binding specificity was verified by competition with unlabeled [Sar1]angiotensin II. Angiotensin II receptor subtypes were characterized by competition with the nonpeptide receptor antagonists, DuP 753 (type 1) and PD123177 (type 2). Both rat and human kidney exhibited a high concentration of angiotensin II receptors in glomeruli and in the longitudinal bands traversing the outer portion of the medulla, corresponding to the medullary vascular bundles. Binding affinity (Kd = 0.6 +/- 0.4 nM), determined in rat kidney, was similar to that reported previously in isolated glomeruli and membrane vesicles prepared from renal tubules. Angiotensin II binding was almost completely inhibited by DuP 753, whereas PD123177 had little effect. Thus the predominant angiotensin II receptor subtype in both rat and human kidney is type 1. The distribution of angiotensin II receptors correlates well with the intrarenal sites at which the peptide has its major physiological effects.
Anoxia induces phospholipase A2 activation in rabbit renal proximal tubules
Authors: Portilla D. , Mandel L. J. , Bar-Sagi D. , Millington D. S.
Published: December 2017

Phospholipase A2 (PLA2) activation during anoxic cell injury was determined by use of a variety of approaches in rabbit proximal renal tubules. Arachidonic acid (AA) mass release increased from 4 +/- 1 (normoxia control) to 40 +/- 6 ng/mg protein after 20 min and 106 +/- 16 ng/mg protein after 40 min of anoxia. PLA2 activity was measured by estimating the amount of sn-2 fatty acid released from either 14C-labeled Escherichia coli membranes or [14C]phosphatidylethanolamine (PE) micelles incubated with membrane and cytosolic fractions obtained from normoxic or anoxic tubules. At pH 7.4 and 1 mM Ca, PLA2 activity increased in the 20-min anoxic membrane fractions from 8.1 +/- 2.3 (normoxic) to 15.2 +/- 2.1 pmol.min-1.mg protein-1 (anoxic). When the proximal tubules were homogenized in the absence of Ca, the anoxia-induced PLA2 activity was found to be soluble. Preincubation with pancreatic PLA2 antibody inhibited 50% of both basal and anoxia-stimulated PLA2 activity. Two protein bands (40- and 21-kDa species) immunoreactive to PLA2 antibody were detected in the membrane fraction. A sixfold increase in the immunoreactivity of the 40-kDa band was detected after 40 min of anoxia of proximal tubules. These results suggest that anoxia induces an intracellular PLA2 activity in kidney cells that could be immunologically related to pancreatic PLA2.
Effect of mesangiolysis on autoregulation of renal blood flow and glomerular filtration rate in rats
Authors: Iversen B. M. , Kvam F. I. , Matre K. , Morkrid L. , Horvei G. , Bagchus W. , Grond J. , Ofstad J.
Published: December 2017

Interlobular arteries and afferent arterioles are involved in autoregulation of renal blood flow (RBF) and glomerular filtration rate (GFR). The question of whether the contractile mesangial cells are also involved in autoregulation was investigated in Wistar rats. Autoregulation of RBF was examined before and 1 h after infusion of antithymocyte (anti-Thy 1-1) antibodies, and both RBF and GFR autoregulation were examined 30 h after the infusion of antibodies. Mesangial cell destruction was present 30 h after the infusion of antibodies. The angiotensin II-induced contraction of isolated glomeruli (70% of control volume, P less than 0.001) was abolished after the glomeruli had been exposed to anti-Thy 1-1 in vitro. RBF, as well as the lower limit of RBF autoregulation, were not different from control 30 h after the infusion (82 +/- 5 vs. 79 +/- 4 mmHg, P greater than 0.10). Autoregulation of GFR was maintained in the control group but was restricted in the experimental group (autoregulatory index: 0.71 +/- 0.42 for left kidney, 0.02 +/- 0.35 for control; P less than 0.05). The afferent arteriolar diameter was unchanged 30 h after the infusion of antibodies (17.8 +/- 0.8 vs. 17.6 +/- 0.4 microns, P greater than 0.10). One hour after infusion of the antibodies, RBF autoregulation was normal. It is concluded that mesangial cells do not seem to be involved in RBF autoregulation, but may in part influence autoregulation of GFR during pressure reduction.
Ion transport by primary cultures of Mongolian gerbil middle ear epithelium
Authors: Herman P. , Friedlander G. , Huy P. T. , Amiel C.
Published: December 2017

The transport properties of Mongolian gerbil middle ear epithelial cells grown in primary culture were studied. These cells formed polarized monolayers that exhibited domes on nonporous supports. On porous supports, monolayers developed an apical-negative transepithelial electric potential difference (VT = -37.2 +/- 2.7 mV) and a transepithelial resistance (RT = 519 +/- 56 omega.cm2). The short-circuit current equivalent (Ieq) was 62.4 +/- 6.2 microA/cm2 (mean +/- SE, n = 15). Na+ and Cl- accumulated in the basal bath and generated a basolateral hyperosmolarity that drove a net water flow. Amiloride (10 microM), when added to the apical but not to the basal bath, induced a 23.4 +/- 1.5 mV and 44.1 +/- 1.3 microA/cm2 decrease of VT and Ieq, respectively, while RT increased by 403 +/- 69 omega.cm2 (P less than 0.001, n = 15). Exposure of the monolayers to a low-Cl- solution (30 mM) enhanced the transepithelial potential, possibly by means of a Cl- secretion through apical Cl- channels. Isoproterenol (10(-4) M basolateral) increased intracellular adenosine 3',5'-cyclic monophosphate (cAMP) content (concentration of half-maximal response = 2.5 x 10(-7) M) and decreased VT, RT, and Ieq. The isoproterenol-induced fall of VT occurred even in the presence of low-Cl-solutions. This suggested an increase of the paracellular pathway conductance, although there was no significant modification of the mannitol permeability.(ABSTRACT TRUNCATED AT 250 WORDS)
Gene expression of growth-related proteins and ECM constituents in response to unilateral nephrectomy
Authors: Nakamura T. , Ebihara I. , Tomino Y. , Koide H. , Kikuchi K. , Koiso K.
Published: December 2017

To identify the specific regulatory mechanism associated with the events following unilateral nephrectomy, we measured the levels of mRNA encoding for extracellular matrix (ECM) constituents, for protooncogenes, and for proliferating cell nuclear antigen (PCNA) in renal cortex and glomeruli. One hour after left nephrectomy, c-jun and c-fos mRNA levels in renal cortex increased rapidly and then decreased rapidly to the control level, whereas c-myc and PCNA mRNA levels showed a slower and more sustained increase, with a peak at 6 h after nephrectomy, and then decreased to the control level after 7 days. mRNA levels for basement membrane components including alpha 1-chain of type IV collagen, laminin B1 and B2 chains, and heparan sulfate proteoglycan core protein were significantly increased in renal cortex at 12 h after nephrectomy, whereas those for interstitial collagens including alpha 1-chains of type I and type III collagen were unchanged following nephrectomy. On the other hand, the glomerular expression of all genes examined in this study showed little change during the experimental period. These results suggest that the time course of mRNA expression of ECM constituents is different from that of growth-related proteins in renal cortex and that glomerular mRNA levels for these components may not be associated with renal hypertrophy in the early stages following unilateral nephrectomy.
Transport properties of an in situ jejunal reservoir in dogs
Authors: Kwon E. D. , Schedl H. P. , Donovan J. F. , Carson R. A. , Williams R. D.
Published: December 2017

To evaluate partial or total replacement of renal function using gut, we measured in vivo transport of nitrogen metabolites, electrolytes, and water into a jejunal segment configured as a continent reservoir in the dog. Reservoir contents were sampled and analyzed at serial time intervals during a 3-h period after instillation of solution containing (in mM) 40 NaCl, 10 NaHCO3, 220 mannitol, pH 8.5, without or with added urease. At 10 min postinstillation, the amount of urea in the solution without added urease was 3-5 times greater than in the presence of added urease, but accumulation of NH4+ was 14-21 times greater in the solution containing added urease, giving a luminal NH4+ concentration up to 10,000 times that of plasma. In the absence of urease, HCO3- concentration fell to 0, and pH declined to 6 at 3 h; in the presence of urease, HCO3- concentration was 4.5 mM, and pH was 7.8 at 3 h. We conclude 1) urea is secreted by the reservoir; 2) H+ is secreted and/or formed in the reservoir; 3) in the presence of urease, urea hydrolyzed to NH3 is converted to NH4+ by H+ and trapped in the lumen; and 4) in the urease solution, H+ binding by NH3 preserves luminal HCO3-, maintaining the initial pH. Thus the continent jejunal reservoir may supplement or replace impaired renal function.
Effect of metabolic acidosis and alkalosis on NEM-sensitive ATPase in rat nephron segments
Authors: Khadouri C. , Marsy S. , Barlet-Bas C. , Cheval L. , Doucet A.
Published: December 2017

An N-ethylmaleimide (NEM)-sensitive adenosinetriphosphatase (ATPase) displaying the kinetic and pharmacological properties of an electrogenic proton pump has been described in the different segments of rat nephron, where it mediates part of the active tubular proton secretion. This study was therefore designed to evaluate whether changes in urinary acidification observed during metabolic acidosis or alkalosis were associated with alterations of the activity of tubular NEM-sensitive ATPase, and if so, to localize the nephron segments responsible for these changes. Within 1 wk after the onset of ammonium chloride treatment, rats developed a metabolic acidosis, and NEM-sensitive ATPase activity was markedly increased in the medullary thick ascending limb of Henle's loop and outer medullary collecting tubule, and slightly increased in the cortical collecting tubule. Conversely, treatment with sodium bicarbonate induced a metabolic alkalosis that was accompanied by decreased NEM-sensitive ATPase activity in medullary thick ascending limb and outer medullary collecting tubule. NEM-sensitive ATPase activity was not altered in any other nephron segment tested in alkalotic and acidotic rats, i.e., the proximal tubule and the cortical thick ascending limb of Henle's loop. Changes qualitatively similar were observed as soon as 3 h after the onset of NaHCO3 or NH4Cl-loading. In the medullary collecting tubule, alterations of NEM-sensitive ATPase activity are in part due to hyperaldosteronism observed in both acidotic and alkalotic rats.(ABSTRACT TRUNCATED AT 250 WORDS)
Renal cytochrome P-450-arachidonic acid metabolism: localization and hormonal regulation in SHR
Authors: Omata K. , Abraham N. G. , Schwartzman M. L.
Published: December 2017

Epoxygenase and omega- and omega-1-hydroxylases are the major cytochrome P-450-arachidonate (P-450-AA) metabolizing enzymes in renal tissues. We measured P-450-AA metabolism in single nephron segments and determined the tubular localization of this activity in spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY). Formation of 20-hydroxyeicosatetraenoic acid (20-HETE), the product of AA omega-hydroxylase was specifically localized in the entire proximal tubules (S1, S2, and S3 segments), whereas formation of 19-HETE, the product of omega-1-hydroxylase and epoxyeicosatrienoic acids (EETs), products of AA epoxygenase, was demonstrable throughout the tubule. Although distribution patterns were similar in SHR and WKY, formation of 19- and 20-HETE in the proximal tubules was higher in SHR, whereas the formation of EETs was not different between the two strains. In the proximal tubules, angiotensin II (ANG II) significantly stimulated epoxygenase activity (EETs formation), whereas parathyroid hormone (PTH) and epidermal growth factor (EGF) had no effect on epoxygenase but significantly stimulated omega-hydroxylase activity (20-HETE formation). Because P-450-AA metabolites have a wide and contrasting spectrum of biological and renal effects, from vasodilation to vasoconstriction and from inhibition to stimulation of Na(+)-K(+)-adenosinetriphosphatase, their localization to the specific nephron segments and differential stimulation of their formation by ANG II, PTH, and EGF may contribute not only to renal hemodynamics and blood pressure regulation but also to the regulation of renal sodium and water balance.
Apical membrane potassium channels in frog diluting segment: stimulation by furosemide
Authors: Hurst A. M. , Hunter M.
Published: December 2017

The patch-clamp technique was used to study the activity of apical membrane potassium channels in frog isolated everted diluting segments, and the effect of transport inhibitors on channel activity was assessed. In cell-attached patches with a high-potassium pipette solution and Ringer in the bath the channels show inward rectification (inward conductance, 25.1 pS; outward conductance, 10.5 pS). The channel is selective for potassium over sodium and is voltage dependent with depolarization increasing channel open probability (Po). Furosemide increased channel activity, which resulted exclusively from a significant increase in the number (N) of channels in the patch (control, 2.3 +/- 0.3, n = 8; furosemide, 4.0 +/- 0.4, n = 14) without any significant change in Po. Amiloride blocked the stimulatory effect of furosemide by reducing N to 1.4 +/- 0.6 (n = 6), and amiloride alone also reduced N with no significant change in Po. This suggests that the increase in N in response to furosemide may be secondary to a rise in intracellular pH mediated by activation of the apical Na-H exchanger.
Renal and systemic effects of urodilatin in rats with high-output heart failure
Authors: Abassi Z. A. , Powell J. R. , Golomb E. , Keiser H. R.
Published: December 2017

Urodilatin is a recently discovered natriuretic peptide [ANP-(95-126)] of renal origin, with a primary structure similar to ANP-(99-126). However, urodilatin is not biologically inactivated by renal endopeptidase, and it is a more potent natriuretic agent than ANP-(99-126). The present study was carried out to investigate the renal and systemic effects of urodilatin in rats before and after the induction of congestive heart failure (CHF) by creation of an aortocaval fistula (ACF). Administration of urodilatin in incremental doses (0.75-12 micrograms.kg-1.h-1) to Inactin-anesthetized sham-operated control rats resulted in dose-dependent increases in urine flow, glomerular filtration rate (GFR), excretion of guanosine 3',5'-cyclic monophosphate (cGMP), sodium, and potassium, and a significant decrease in mean arterial blood pressure. In rats with ACF the baseline values for GFR and sodium excretion were significantly lower than in control rats. Urodilatin infusion in rats with ACF led to significant increases in urine flow and sodium excretion, but the absolute levels of diuresis and natriuresis were significantly lower in rats with CHF than in normal rats. When urodilatin was infused into rats with ACF pretreated with neutral endopeptidase inhibitor (NEP-I; SQ-28,063 at a dose of 40 mg/kg iv), the absolute urine flow and sodium excretion were not different from that obtained in control rats. Thus the attenuated natriuretic and diuretic response to ANP-(99-126) in heart failure was not observed with urodilatin.(ABSTRACT TRUNCATED AT 250 WORDS)
Effect of dietary protein on renin and angiotensinogen gene expression after renal ablation
Authors: Correa-Rotter R. , Hostetter T. H. , Rosenberg M. E.
Published: December 2017

High dietary protein increases renin mRNA and likely contributes to the increased plasma renin activity (PRA) in the normal rat. In experimental renal ablation, dietary protein influences renal growth and injury. This study was undertaken to examine the effect of dietary protein on the renin-angiotensin system after reduction of renal mass. Male Sprague-Dawley rats were subjected to 1 1/3 nephrectomy (1 1/3 NX) and maintained for 7 days on a low-protein (6%) diet, at which time they were stratified according to serum creatinine to either continue on the low-protein diet or were switched to a high-protein (40%) diet. Two weeks after stratification renal renin mRNA was higher on the 40% compared with the 6% protein diet, angiotensinogen mRNA was lower in the high-protein group, and no change occurred in renal actin mRNA. The magnitude of this dietary protein-induced increase in renin mRNA with high-protein diet was greater in the 1 1/3 NX group compared with that observed in sham-operated rats (320 vs. 60%). PRA and tissue renin activity were also elevated on the high-protein diet after 1 1/3 NX. When three levels of dietary protein intake were examined after 1 1/3 NX, the high-protein (40%) diet was associated with increased renal renin mRNA compared with both the intermediate-protein (20%) and low-protein (6%) diets. In conclusion, a high-protein diet induced activation of the renin-angiotensin system. This activation may explain some of the detrimental effects of dietary protein on the course of chronic renal failure.
Multiple signaling pathways for Cl(-)-dependent depolarization of mesangial cells: role of Ca2+, PKC, and G proteins
Authors: Kremer S. G. , Zeng W. , Sridhara S. , Skorecki K. L.
Published: December 2017

We have previously reported that vasopressin activates chloride channels, leading to depolarization of glomerular mesangial cells via both calcium-dependent and calcium-independent pathways (S.G. Kremer, W.V. Breuer, and K. L. Skorecki, J. Cell. Physiol. 138: 97-105, 1989). However, the calcium-independent pathways were not defined. Using fluorescent probes, we now demonstrate that activation of protein kinase C (PKC) results in cellular depolarization in the absence of a calcium signal. This depolarization is also mediated by an enhanced conductance to chloride. Downregulation of PKC partially attenuated but did not abolish the depolarization response to vasopressin. Depolarization persisted when, in addition, calcium responses were also abolished and prostaglandin production was eliminated, suggesting an additional pathway for depolarization. G protein activation by aluminum fluoride also resulted in cellular depolarization mediated by an enhanced conductance to chloride, which persisted when calcium and PKC-signaling pathways were eliminated. This suggests the presence of a calcium- and PKC-independent pathway for G protein-mediated chloride-dependent depolarization. These findings point to the presence of at least three separate signaling pathways available for the activation of mesangial cell chloride channels, i.e., calcium, PKC, and a G protein.
Integrin receptors in the glomerulus: potential role in glomerular injury
Authors: Adler S.
Published: December 2017

The extracellular matrix (ECM) exerts important effects on cell function that play a vital role in such diverse processes as embryogenesis, wound healing, cancer metastasis, and inflammation. Interactions between cells and matrix are mediated by a variety of cell surface receptors for different components of the ECM. The integrin family of matrix receptors, the best-studied group thus far, has already been shown to play an important role in cell-matrix and cell-cell interactions in other systems. More recently, the types of integrin receptors present in the glomerulus and on cultured glomerular cells have been defined, and their potential involvement in glomerular injury has been postulated. This review will summarize current knowledge of integrin receptors, their distribution in the kidney, cultured glomerular cell-matrix interactions that might play a role in modulating the glomerular cell response to injury, and the potential role of integrins and other glomerular cell matrix receptors as targets of injury in proteinuric lesions.
Blockade of pressure natriuresis induced by inhibition of renal synthesis of nitric oxide in dogs
Authors: Salom M. G. , Lahera V. , Miranda-Guardiola F. , Romero J. C.
Published: December 2017

To evaluate the participation of nitric oxide (NO) on pressure-induced natriuresis in pentobarbital-anesthetized dogs, renal perfusion pressure (RPP) was increased twice from 100 to 150 mmHg before and during the intrarenal administration of an NO-synthesis inhibitor, NG-nitro-L-arginine methyl ester (L-NAME), while determining changes in glomerular filtration rate (GFR), renal blood flow (RBF), and urine sodium and water excretion. Before the inhibition of NO, the increase in RPP induced diuresis (5-fold) and natriuresis (4.2-fold) with no change in RBF or GFR. However, the intrarenal infusion of L-NAME (1 microgram.kg-1.min-1) blunted the diuretic and natriuretic responses without altering RBF or GFR. The infusion of the NO synthesis precursor L-arginine prevented the inhibitory effect that L-NAME exerted on the diuretic and natriuretic responses to the increase in RPP. These results indicate that the increase in RPP stimulates NO synthesis and suggest that NO might play an important role in the control of sodium and water excretion during acute changes in RPP.