Cell polarity and calcium oxalate crystal adherence to cultured collecting duct cells

1992 ◽  
Vol 262 (2) ◽  
pp. F177-F184 ◽  
Author(s):  
R. J. Riese ◽  
N. S. Mandel ◽  
J. H. Wiessner ◽  
G. S. Mandel ◽  
C. G. Becker ◽  
...  

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.

1985 ◽  
Vol 248 (4) ◽  
pp. F500-F506 ◽  
Author(s):  
B. M. Koeppen

Segments of outer medullary collecting duct were dissected from the inner stripe of the rabbit kidney (OMCDi) and perfused in vitro. The conductive properties of the tubule epithelium and individual cell membranes were determined by means of cable analysis and intracellular voltage-recording microelectrodes. In 35 tubules the transepithelial voltage (VT) and resistance (RT) averaged 17.2 +/- 1.4 mV, lumen positive, and 58.6 +/- 5.3 k omega X cm, respectively. The basolateral membrane voltage, (Vbl) was -29.2 +/- 2.1 mV (n = 23). The apical cell membrane did not contain appreciable ion conductances, as evidenced by the high values of apical cell membrane fractional resistance (fRa = Ra/Ra + Rb), which approached unity (0.99 +/- 0.01; n = 23). Moreover, addition of amiloride or BaCl2 to the tubule lumen was without effect on the electrical characteristics of the cell, as was a twofold reduction in luminal [Cl-]. The conductive properties of the basolateral cell membrane were assessed with bath ion substitutions. A twofold reduction in bath [Cl-] depolarized Vbl by 14.7 +/- 0.4 mV (theoretical, 17 mV), while a 10-fold increase in bath [K+] resulted in only a 0.9 +/- 0.4 mV depolarization (theoretical, 61 mV). Substituting bath Na+ with tetramethylammonium (from 150 to 75 mM) was without effect. Reducing bath [HCO-3] from 25 to 5 mM (constant PCO2) resulted in a steady-state depolarization of Vbl of 8.4 +/- 0.4 mV that could not be attributed to conductive HCO-3 movement. Thus, the basolateral cell membrane is predominantly Cl- selective.(ABSTRACT TRUNCATED AT 250 WORDS)


1982 ◽  
Vol 60 (9) ◽  
pp. 1149-1152 ◽  
Author(s):  
H. Sonnenberg ◽  
W. A. Cupples ◽  
A. J. de Bold ◽  
A. T. Veress

In anesthetized rats micropuncture and microcatheterization were used to collect tubular fluid from end proximal and distal tubules and from the outer medullary collecting duct. Urine was collected at the papilla tip. Samples were taken from the same sites before and after intravenous injection of atrial tissue extract; rats injected with ventricular extract served as controls. Sodium excretion increased 17-fold after atrial extract, a significantly greater rise than the 3-fold increase after ventricular extract. Clearances of inulin and single nephron filtration rates did not change significantly in either group. Tubular fluid collection results showed a similar reduction (16 to 20%) of proximal fluid and sodium reabsorption in both groups. In the experimental group only, NaCl reabsorption failed to rise in response to increased load in the medullary collecting duct. The resulting fall in fractional reabsorption in the medullary collecting duct accounted for 80% of the natriuresis. We conclude that atrial tissue from rat hearts contains a factor which causes increased renal NaCl excretion by inhibiting transport in the medullary collecting duct.


1996 ◽  
Vol 271 (2) ◽  
pp. F322-F329 ◽  
Author(s):  
H. Wald ◽  
O. Goldstein ◽  
C. Asher ◽  
Y. Yagil ◽  
H. Garty

CHIF is a recently cloned, corticosteroid-induced gene which evokes K+ channel activity in oocytes (B. Attali, H. Latter, N. Rachamim, and H. Garty. Proc. Natl. Acad. Sci. USA 92: 6092-6096, 1995). To further characterize the possible role of this gene in epithelial ion transport, we have studied its epithelial distribution and hormonal induction. Northern hybridizations indicate that the zonal distribution of CHIF mRNA in kidney is: papilla >>medulla>> cortex. High levels of CHIF were also detected in a primary culture from inner medullary collecting duct (IMCD). Perfusing rats with < 20 nM aldosterone through osmotic minipumps evoked a 22.4 +/- 1.9-fold increase in colonic CHIF. A significant increase was observed 3 h after administrating the corticosteroid, but maximal response was detected only after a 72-h incubation. This response appears to be mineralocorticoid specific; perfusing or injecting rats with maximal doses of dexamethasone did not evoke a further increase in CHIF mRNA. In contrast, high levels of CHIF are expressed in kidney papilla and IMCD primary culture, irrespective of corticosteroid treatment. Thus, like the apical Na+ channel and the H(+)-K(+)-adenosinetriphosphatase, CHIF is mineralocorticoid induced in the colon but constitutively expressed in kidney.


1990 ◽  
Vol 258 (2) ◽  
pp. F266-F272 ◽  
Author(s):  
S. P. Nadler

To assess the effects of increased tonicity on water reabsorption (Jv) in inner medullary collecting ducts (IMCD), antidiuretic hormone (ADH)-stimulated Jv and water permeability (PF) were determined in microperfused IMCD dissected from the inner medulla of rat kidney. In IMCD exposed to a 150-mosmol/kgH2O gradient in isotonic bath, ADH-stimulated PF averaged 719 +/- 93 microns/s. Symmetric addition of 75 mM NaCl to perfusate and bath resulted in a significant augmentation of ADH-stimulated PF (56%) that was reversible when initial solutions were restored. Despite the increase in PF, JV did not change but would have decreased by 16% (P less than 0.01) had PF not increased, because of the greater absolute axial increase in luminal tonicity that occurs with more hypertonic luminal solutions. When 150 mM mannitol was used to increase tonicity, similar effects were observed. However, 150 mM urea had no effect on ADH-stimulated PF. In IMCD exposed to 8-para-(chlorophenylthio)-adenosine 3',5'-cyclic monophosphate, addition of 75 mM NaCl to both and perfusate also resulted in a 76% increase in PF. These results are the first to demonstrate directly that increased effective tonicity augments ADH-stimulated PF in rat IMCD at a site distal to adenosine 3',5'-cyclic monophosphate generation. This effect may contribute to maintenance of medullary interstitial tonicity during antidiuresis by ensuring that most water reabsorption occurs more proximally within the IMCD.


1990 ◽  
Vol 1 (6) ◽  
pp. 890-901
Author(s):  
D Kikeri ◽  
M L Zeidel

To define proton transport mechanisms involved in the regulation of intracellular pH (pHi) in cells of the inner medullary collecting duct (IMCD), pHi and cell membrane potential were estimated by using the fluorescent dyes 2,7-biscarboxyethyl-5(6)-carboxyfluorescein and 3,3'-dipropylthiadicarbocyanine iodide, respectively, in suspensions of freshly isolated rabbit IMCD cells. The resting pHi of IMCD cells in nonbicarbonate Ringer's solution (pH 7.4) was 7.21 +/- 0.03 (mean +/- SE). When cells were acidified by ammonium withdrawal, the initial pHi recovery rate was 0.33 +/- 0.02 pH unit/min; replacement of extracellular Na+ (130 mM) with N-methyl-D-glucamine+ reduced the pHi recovery rate to 0.08 +/- 0.02 pH unit/min, while addition of 0.1 mM amiloride in the presence of extracellular Na+ reduced the rate of pHi recovery to 0.02 +/- 0.02 pH unit/min. Similar results were obtained in cells acid loaded with HCl. Cells recovering from acidification exhibited 22Na+ uptake rates threefold higher than did nonacidified cells. The rate of Na(+)-dependent pHi recovery was independent of the cell membrane potential. In the absence of extracellular Na+, depolarizing cell membrane potential in a stepwise manner by increasing extracellular K+ concentrations from 1 to 130 mM resulted in graded increments in the rate of pHi recovery. In the presence of 130 mM K+, the pHi recovery rate in acidified cells was dependent on cellular ATP levels, sensitive to 1 mM N-ethylmaleimide, and insensitive to 0.01 mM oligomycin in the presence of glucose (control, 0.24 +/- 0.01; ATP-depleted, 0.13 +/- 0.02; addition of N-ethylmaleimide, 0.16 +/- 0.01; addition of oligomycin, 0.27 +/- 0.02 pH unit/min). ATP depletion markedly inhibited H+ extrusion from IMCD cells measured by using a pH stat. These results provide direct evidence in freshly isolated IMCD cells that both a Na+:H+ antiporter and a rheogenic H(+)-ATPase participate in pHi regulation.


1998 ◽  
Vol 274 (5) ◽  
pp. F841-F855 ◽  
Author(s):  
Alan M. Weinstein

A mathematical model of the inner medullary collecting duct (IMCD) of the rat has been developed representing Na+, K+, Cl−,[Formula: see text] CO2, H2CO3, phosphate, ammonia, and urea. Novel model features include: finite rates of hydration of CO2, a kinetic representation of the H-K-ATPase within the luminal cell membrane, cellular osmolytes that are regulated in defense of cell volume, and the repeated coalescing of IMCD tubule segments to yield the ducts of Bellini. Model transport is such that when entering Na+ is 4% of filtered Na+, approximately 75% of this load is reabsorbed. This requirement renders the area-specific transport rate for Na+ comparable to that for proximal tubule. With respect to the luminal membrane, there is experimental evidence for both NaCl cotransport and an Na+ channel in parallel. The experimental constraints that transepithelial potential difference is small and that the fractional apical resistance is greater than 85% mandate that more than 75% of luminal Na+ entry be electrically silent. When Na+delivery is limited, an NaCl cotransporter can be effective at reducing luminal Na+ concentration to the observed low urinary values. Given the rate of transcellular Na+ reabsorption, there is necessarily a high rate of peritubular K+recycling; also, given the lower bound on luminal membrane Cl− reabsorption, substantial peritubular Cl− flux must be present. Thus, if realistic limits on cell membrane electrical resistance are observed, then this model predicts a requirement for peritubular electroneutral KCl exit.


1999 ◽  
Vol 277 (1) ◽  
pp. F10-F16 ◽  
Author(s):  
Tianxin Yang ◽  
Daqing Sun ◽  
Yuning G. Huang ◽  
Ann Smart ◽  
Josephine P. Briggs ◽  
...  

Induction of the inducible cyclooxygenase isoform COX-2 is likely to be an important mechanism for increased prostaglandin production in renal inflammation. We examined the effect of lipopolysaccharide (LPS) on regional renal COX-2 expression in the rat. In the inner medulla, LPS injection (4 mg/kg ip) induced a twofold and 2.5-fold increase in the levels of COX-2 mRNA and COX-2 protein, respectively. In contrast, COX-2 expression in the renal cortex was not significantly altered. COX-2 promoter transgenic mice were created using the 2.7-kb flanking region of the rat COX-2 gene. In these animals, LPS injection induced reporter gene expression predominately in the inner medulla. The LPS receptor CD14, usually regarded as a monocyte/macrophage-specific marker, was found to be abundantly expressed in the inner medulla and in dissected inner medullary collecting duct (IMCD) cells, suggesting that it may mediate medullary COX-2 induction. CD14 was present only at low levels in cortex and cortical segments, including glomeruli. In cultured cells, it was abundant in mouse IMCD (mIMCD-K2) cells and renal medullary interstitial cells, but largely undetectable in mesangial cells and M1 cells, a cell line derived from mouse cortical collecting ducts. In the mIMCD-K2 cell line, LPS significantly induced COX-2 mRNA expression, with concomitant induction of CD14. LPS-stimulated COX-2 expression was reduced by the addition of an anti-CD14 monoclonal antibody to the culture medium. These results demonstrate that LPS selectively stimulates COX-2 expression in the renal inner medulla through a CD14-dependent mechanism.


1986 ◽  
Vol 250 (1) ◽  
pp. F70-F76 ◽  
Author(s):  
B. M. Koeppen

Segments of the outer medullary collecting duct were dissected from the outer stripe of the rabbit kidney (OMCDo) and perfused in vitro. The conductive properties of the tubule epithelium and individual cell membranes were determined by means of cable analysis and intracellular voltage-recording microelectrodes. The transepithelial voltage (VT) and resistance (RT) averaged -10.7 +/- 2.5 mV, lumen negative, and 28.5 +/- 2.9 k omega X cm (n = 27), respectively. Two cell types could be defined by their electrophysiological properties. One cell type (n = 7) had a mean basolateral membrane voltage (Vbl) of -30.1 +/- 2.4 mV, a fractional resistance of the apical membrane (fRa = Ra/Ra + Rbl) near unity (0.99 +/- 0.01), and a predominantly Cl(-)-selective basolateral cell membrane. The second cell type (n = 27) had a mean Vbl of -63.7 +/- 2.7 mV, a fRa of 0.81 +/- 0.02, and a predominantly K+-selective basolateral cell membrane. The present study focused on defining the conductive properties of this latter cell type. Amiloride (10(-5) M) and BaCl2 (2 mM) were used as probes of apical cell membrane Na+ and K+ conductive pathways, respectively. Amiloride increased fRa from 0.80 +/- 0.02 to 0.98 +/- 0.01 (n = 12), whereas BaCl2 increased fRa from 0.77 +/- 0.03 to 0.82 +/- 0.03 (n = 9). The conductive properties of the basolateral cell membrane were assessed by ion substitutions of the bath solution. A 10-fold increase in the bath [K+] depolarized Vbl by 34.9 +/- 1.9 mV (n = 16) in less than 1 s, indicating that this membrane was predominantly K+ selective.(ABSTRACT TRUNCATED AT 250 WORDS)


Urology ◽  
2016 ◽  
Vol 94 ◽  
pp. 47-52 ◽  
Author(s):  
Marcelino Rivera ◽  
Patrick A. Cockerill ◽  
Felicity Enders ◽  
Ramila A. Mehta ◽  
Lisa Vaughan ◽  
...  

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