Postnatal maturation of rabbit renal collecting duct: intercalated cell function

1987 ◽  
Vol 253 (4) ◽  
pp. F622-F635 ◽  
Author(s):  
L. M. Satlin ◽  
G. J. Schwartz
Keyword(s):  
Fluorescent Dyes ◽  
Cell Function ◽  
Collecting Duct ◽  
Intercalated Cells ◽  
Mitochondrial Potential ◽  
Postnatal Maturation ◽  
Intercalated Cell ◽  
Cell Ph ◽  
Cytoplasmic Vesicles ◽  
Collecting Ducts

Intercalated cells play a major role in renal regulation of acid-base balance. We used fluorescent dyes to characterize postnatal maturation of intercalated cells. We stained rabbit collecting ducts with the pH-sensitive dye 6-carboxyfluorescein diacetate and identified individual intercalated cells by their bright green fluorescence. Number of fluorescent cells per millimeter tubule doubled during maturation in midcortex (68 +/- 7 to 121 +/- 9; P less than 0.01) but did not change in outer stripe of outer medulla. Excitation-ratio (490/450 nm) fluorometry of individual cells in nonperfused tubules revealed an increase in pH of cortical intercalated cell from 7.28 +/- 0.03 in newborn to 7.43 +/- 0.03 in adult (P less than 0.005); principal cell pH did not change with age, averaging 7.10 +/- 0.05. The smaller difference in pH between intercalated and principal cells in neonates suggested a paucity of H+ pumps in immature intercalated cells. Indeed, few cortical intercalated cells trapped the weak base acridine orange in cytoplasmic vesicles that contained H+ pumps or demonstrated selective uptake of 3,3'+-dipentyloxacarbocyanine, a fluorescent cation that probes for mitochondrial potential. Intercalated cells in neonatal medullary collecting ducts had a cell pH similar to that measured in the adult, as well as numerous acidic cytoplasmic vesicles and significant mitochondrial potentials. In conclusion, intercalated cells in cortical collecting duct underwent postnatal proliferation and maturation, whereas those cells present in outer medullary collecting duct, where proliferation was virtually complete by 1 wk of age, were nearly differentiated. Signals directing this centrifugal pattern of postnatal renal maturation are presently unknown.

Postnatal maturation of rabbit renal collecting duct. III. Peanut lectin-binding intercalated cells

1992 ◽  
Vol 262 (2) ◽  
pp. F199-F208 ◽  
Author(s):  
L. M. Satlin ◽  
T. Matsumoto ◽  
G. J. Schwartz
Keyword(s):  
Collecting Duct ◽  
Lectin Binding ◽  
Rabbit Kidney ◽  
Intercalated Cells ◽  
Outer Cortex ◽  
Cell Surface Antigens ◽  
Postnatal Maturation ◽  
Intercalated Cell ◽  
Cell Ph ◽  
Alkaline Cell

Measurements of transepithelial HCO3 transport in the rabbit cortical collecting duct (CCD) indicate that net HCO3 secretion becomes apparent only after the first month of life [F. M. Mehrgut, L. M. Satlin, and G. J. Schwartz, Am. J. Physiol. 259 (Renal Fluid Electrolyte Physiol. 28): F801-F808, 1990]. We used fluorescent probes and immunocytochemistry to trace the postnatal functional development of the beta-intercalated cell, the HCO3-secreting cell of the fully differentiated CCD. Throughout maturation, the beta-intercalated cell was empirically identified by its selective uptake of the pH-sensitive dye 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein, an alkaline cell pH, apical binding to peanut agglutinin (PNA) and monoclonal antibody B63, and by its functional capacity for apical Cl-HCO3 exchange as manifested by Cl-dependent extrusion of an intracellular alkali load. Compared with the mature segment, the neonatal mid-CCD exhibited fewer intercalated cells, which were characterized by a less alkaline cell pH, reduced apical Cl-HCO3 exchange activity, and shorter apical binding profiles for PNA. There was evidence for basolateral Cl conductance and similar buffering capacity at all ages. In the neonatal outer cortex there was little or no binding to PNA or to B63. As soon as cell surface antigens characteristic of the fully differentiated beta-cell were detected, functional studies indicated the presence, albeit reduced, of apical Cl-HCO3 exchange. Thus there is postnatal proliferation and maturation of HCO3-secreting intercalated cells in the rabbit kidney; the origin of these cells remains to be determined.

Lectin-gold cytochemistry reveals intercalated cell heterogeneity along rat kidney collecting ducts

1985 ◽  
Vol 248 (3) ◽  
pp. C348-C356 ◽  
Author(s):  
D. Brown ◽  
J. Roth ◽  
L. Orci
Keyword(s):  
Plasma Membranes ◽  
Collecting Duct ◽  
Rat Kidney ◽  
Lectin Binding ◽  
Helix Pomatia ◽  
Intercalated Cells ◽  
Double Labeling ◽  
Thin Sections ◽  
Intercalated Cell ◽  
Collecting Ducts

The lectin-gold technique was used to detect Helix pomatia and Dolichos biflorus lectin binding sites directly on semithin and thin sections of rat kidney collecting ducts. Intercalated cell apical plasma membranes and the membranes of apical cytoplasmic vesicles were heavily labeled in the cortex and outer stripe of the outer medulla but were negative or very weakly labeled in the inner stripe and inner medulla. In contrast, clear cell apical membranes were labeled along the entire length of the collecting duct. Double labeling of semithin cryostat sections with a specific antibody and lectin-gold complexes was used to demonstrate that the intercalated cells in all regions studied contained carbonic anhydrase, even though the lectin binding differed. These results indicate that, in terms of their glycocalyx composition, intercalated cells represent a heterogeneous population in different regions of the collecting duct.

Mechanisms of HCO 3 − secretion in the rabbit connecting segment

1999 ◽  
Vol 277 (4) ◽  
pp. F567-F574 ◽  
Author(s):  
Shuichi Tsuruoka ◽  
George J. Schwartz
Keyword(s):  
Cell Function ◽  
Collecting Duct ◽  
Intercalated Cells ◽  
Cortical Collecting Duct ◽  
Acid Base ◽  
Intercalated Cell ◽  
Atpase Inhibitors ◽  
Transepithelial Voltage ◽  
Β Adrenergic Receptors

The connecting tubule (CNT) contains α-(H+-secreting) and β-([Formula: see text]-secreting) intercalated cells and is therefore likely to contribute to acid-base homeostasis. To characterize the mechanisms of [Formula: see text]transport in the rabbit CNT, in which there is little definitive data presently available, we microdissected the segments from the superficial cortical labyrinth, perfused them in vitro, measured net[Formula: see text] transport ( J [Formula: see text] ) by microcalorimetry, and examined the effects of several experimental maneuvers. Mean ± SE basal J [Formula: see text] was −3.4 ± 0.1 pmol ⋅ min−1 ⋅ mm−1(net [Formula: see text] secretion), and transepithelial voltage was −13 ± 1 mV ( n = 47). Net[Formula: see text] secretion was markedly inhibited by removal of luminal Cl− or application of basolateral H+-ATPase inhibitors (bafilomycin or concanamycin), maneuvers that inhibit β-intercalated cell function. Net [Formula: see text] secretion was not affected by inhibitors of α-intercalated cell function (basolateral Cl− removal, basolateral DIDS, or luminal H+-ATPase inhibitors). Net [Formula: see text] secretion was stimulated by isoproterenol and inhibited by acetazolamide. These data indicate that 1) CNTs secrete[Formula: see text] via an apical DIDS-insensitive Cl−/[Formula: see text]exchanger, mediated by a basolateral bafilomycin- and concanamycin-sensitive H+-ATPase; 2) inhibition of cytosolic carbonic anhydrase decreases [Formula: see text] secretion; and 3) stimulation of β-adrenergic receptors increases [Formula: see text] secretion. The failure to influence net [Formula: see text]transport by inhibiting α-intercalated cell apical H+-ATPases or basolateral Cl−/[Formula: see text]exchange suggests that the CNT has fewer functioning α-intercalated cells than the cortical collecting duct. These are the first studies to examine the rate and mechanisms of[Formula: see text] secretion by the rabbit CNT; this is clearly an important segment in mediating acid-base homeostasis.

scholarly journals Potassium depletion increases proton pump (H+-ATPase) activity in intercalated cells of cortical collecting duct

2000 ◽  
Vol 279 (1) ◽  
pp. F195-F202 ◽  
Author(s):  
Randi B. Silver ◽  
Sylvie Breton ◽  
Dennis Brown
Keyword(s):  
Plasma Membrane ◽  
Atpase Activity ◽  
Collecting Duct ◽  
Proton Pumps ◽  
Intercalated Cells ◽  
Cortical Collecting Duct ◽  
Collecting Ducts ◽  
Acid Load ◽  
Collecting Tubules ◽  
Atpase Staining

Intercalated cells (ICs) from kidney collecting ducts contain proton-transporting ATPases (H+-ATPases) whose plasma membrane expression is regulated under a variety of conditions. It has been shown that net proton secretion occurs in the distal nephron from chronically K+-depleted rats and that upregulation of tubular H+- ATPase is involved in this process. However, regulation of this protein at the level of individual cells has not so far been examined. In the present study, H+-ATPase activity was determined in individually identified ICs from control and chronically K+-depleted rats (9–14 days on a low-K+ diet) by monitoring K+- and Na+-independent H+ extrusion rates after an acute acid load. Split-open rat cortical collecting tubules were loaded with the intracellular pH (pHi) indicator 2′,7′-bis(2-carboxyethyl)-5(6)-carboxyfluorescein, and pHiwas determined by using ratiometric fluorescence imaging. The rate of pHi recovery in ICs in response to an acute acid load, a measure of plasma membrane H+-ATPase activity, was increased after K+ depletion to almost three times that of controls. Furthermore, the lag time before the start of pHirecovery after the cells were maximally acidified fell from 93.5 ± 13.7 s in controls to 24.5 ± 2.1 s in K+-depleted rats. In all ICs tested, Na+- and K+-independent pHi recovery was abolished in the presence of bafilomycin (100 nM), an inhibitor of the H+-ATPase. Analysis of the cell-to-cell variability in the rate of pHi recovery reveals a change in the distribution of membrane-bound proton pumps in the IC population of cortical collecting duct from K+-depleted rats. Immunocytochemical analysis of collecting ducts from control and K+-depleted rats showed that K+-depletion increased the number of ICs with tight apical H+ATPase staining and decreased the number of cells with diffuse or basolateral H+-ATPase staining. Taken together, these data indicate that chronic K+ depletion induces a marked increase in plasma membrane H+ATPase activity in individual ICs.

scholarly journals Intercalated cell-specific Rh B glycoprotein deletion diminishes renal ammonia excretion response to hypokalemia

2013 ◽  
Vol 304 (4) ◽  
pp. F422-F431 ◽  
Author(s):  
Jesse M. Bishop ◽  
Hyun-Wook Lee ◽  
Mary E. Handlogten ◽  
Ki-Hwan Han ◽  
Jill W. Verlander ◽  
...  
Keyword(s):  
Metabolic Acidosis ◽  
Collecting Duct ◽  
Ammonia Excretion ◽  
Ammonia Concentration ◽  
Intercalated Cells ◽  
Cortical Collecting Duct ◽  
Intercalated Cell ◽  
Transporter Family ◽  
Total Ammonia ◽  
Normal Increase

The ammonia transporter family member, Rh B Glycoprotein (Rhbg), is an ammonia-specific transporter heavily expressed in the kidney and is necessary for the normal increase in ammonia excretion in response to metabolic acidosis. Hypokalemia is a common clinical condition in which there is increased renal ammonia excretion despite the absence of metabolic acidosis. The purpose of this study was to examine Rhbg's role in this response through the use of mice with intercalated cell-specific Rhbg deletion (IC-Rhbg-KO). Hypokalemia induced by feeding a K+-free diet increased urinary ammonia excretion significantly. In mice with intact Rhbg expression, hypokalemia increased Rhbg protein expression in intercalated cells in the cortical collecting duct (CCD) and in the outer medullary collecting duct (OMCD). Deletion of Rhbg from intercalated cells inhibited hypokalemia-induced changes in urinary total ammonia excretion significantly and completely prevented hypokalemia-induced increases in urinary ammonia concentration, but did not alter urinary pH. We conclude that hypokalemia increases Rhbg expression in intercalated cells in the cortex and outer medulla and that intercalated cell Rhbg expression is necessary for the normal increase in renal ammonia excretion in response to hypokalemia.

Three distinct cell populations in rat kidney collecting duct

1987 ◽  
Vol 253 (2) ◽  
pp. C323-C328 ◽  
Author(s):  
H. Holthofer ◽  
B. A. Schulte ◽  
G. Pasternack ◽  
G. J. Siegel ◽  
S. S. Spicer
Keyword(s):  
Collecting Duct ◽  
Rat Kidney ◽  
Morphological Characteristics ◽  
Anion Channel ◽  
Band 3 ◽  
Cell Populations ◽  
Intercalated Cells ◽  
Distinct Cell ◽  
Collecting Ducts ◽  
Principal And Intercalated Cells

The morphologically heterogeneous cell populations in the collecting ducts of the rat kidney were studied using immunocytochemical detection of Na+-K+-ATPase and the anion channel (band 3) glycoprotein. Both enzymes were localized to the basal aspect of separate and morphologically distinct subpopulations of cells in various segments of the collecting duct. Na+-K+-ATPase appeared to be present exclusively in principal cells as identified by their morphology, whereas band 3 antibodies reacted only with intercalated cells. However, 5-20% of cells with the morphological characteristics of intercalated cells failed to react with either antisera in various segments of collecting ducts. As band 3 glycoprotein serves in exchanging intracellular bicarbonate for chloride, it is highly likely that the cells positive for this antigen secrete protons. The method introduced here appears thus useful for distinguishing between principal and intercalated cells by differences in their enzyme content and further for revealing two subpopulations of intercalated cells. This method promises to provide a useful approach for studying the principal and intercalated cell populations in various metabolic states.

Axial heterogeneity of rabbit cortical collecting duct

1991 ◽  
Vol 260 (4) ◽  
pp. F498-F505
Author(s):  
C. L. Emmons ◽  
K. Matsuzaki ◽  
J. B. Stokes ◽  
V. L. Schuster
Keyword(s):  
Cross Sections ◽  
Cell Function ◽  
Rate Coefficient ◽  
Collecting Duct ◽  
Lectin Binding ◽  
Cell Types ◽  
Intercalated Cells ◽  
Cortical Collecting Duct ◽  
Principal Cells ◽  
Inner Cortex

The rabbit cortical collecting duct (CCD) consists of three major cell types: principal cells transport K+, beta-intercalated cells absorb Cl-, and alpha-intercalated cells secrete H+. We used functional and histological methods to assess axial distribution of these cell types along rabbit CCD. In perfused CCDs, lumen-to-bath Rb+ rate coefficient (an index of principal cell K+ transport) was not different in tubules from outer cortex (1 mm from renal surface) compared with those from inner cortex (2 mm from renal surface), suggesting that principal cell function is homogeneous along the CCD. In contrast, Cl- rate coefficient (a measure of beta-intercalated cell function) was twice as high in CCDs from outer compared with inner cortex, suggesting heterogeneity of beta-intercalated cells along the CCD. To further investigate these regional differences, we fixed and embedded kidneys and identified three cell types in CCD cross sections using carbonic anhydrase staining and peanut lectin binding. Comparing tubule cross sections from outer with those from inner cortex, we found no axial difference in the fraction of cells that were either principal cells (64%) or total (lectin binding and nonlectin binding) intercalated cells (36%). However, the lectin-binding intercalated cell subset was significantly increased in outer compared with inner cortex. We conclude that there is not heterogeneity of principal cells along the rabbit CCD; however, beta-cell number and function are increased in outer CCD. Collecting duct heterogeneity begins within the cortical segment.

Activation of acid-secreting intercalated cells in rabbit collecting duct with ammonium chloride loading

1994 ◽  
Vol 266 (4) ◽  
pp. F633-F645 ◽  
Author(s):  
J. W. Verlander ◽  
K. M. Madsen ◽  
J. K. Cannon ◽  
C. C. Tisher
Keyword(s):  
Plasma Membrane ◽  
Morphometric Analysis ◽  
Collecting Duct ◽  
Band 3 ◽  
Apical Plasma Membrane ◽  
Multivesicular Bodies ◽  
Intercalated Cells ◽  
Cytoplasmic Vesicles ◽  
Basolateral Plasma Membrane ◽  
Acid Loading

In normal rabbit, immunolabeling of intercalated cells in the outer medullary collecting duct (OMCD) demonstrates band 3-like protein in the basolateral plasma membrane (15) and H(+)-adenosinetriphosphatase (H(+)-ATPase) in the apical plasma membrane and cytoplasmic vesicles (30). However, in type A intercalated cells in the cortical collecting duct (CCD), band 3-like protein is located primarily in multivesicular bodies and cytoplasmic vesicles (15), whereas H(+)-ATPase is present in cytoplasmic vesicles only in most intercalated cells (30). In this study, we observed the effect of chronic acid loading on immunolocalization of these transporters in the collecting duct. Adult New Zealand White rabbits received either normal tap water (controls) or 75 mM NH4Cl for 12 days plus eight daily gavages of 2-6 meq NH4Cl/kg body wt. At time of death, mean urine pH of acid-loaded animals was 5.96 (SD = 0.69), vs. 8.47 (SD = 0.07) in controls. Kidneys were fixed by in vivo perfusion and processed for light and electron microscopic immunoperoxidase localization of band 3-like protein and immunogold localization of H(+)-ATPase. In controls, band 3-like protein was largely confined to multivesicular bodies in the majority of positive-staining intercalated cells in the CCD and to the basolateral plasma membrane of intercalated cells in the OMCD. In acid-loaded rabbits, band 3 protein-positive intercalated cells in the inner CCD and the in the outer stripe of the OMCD (OMCDo) were strikingly stellate in form. Basolateral plasma membrane label was intensified, while the number of labeled multivesicular bodies was diminished. Morphometric analysis demonstrated an increase in the amount of basolateral plasma membrane in these intercalated cells. In control rabbits, H(+)-ATPase immunoreactivity in intercalated cells in the CCD was located predominantly over cytoplasmic vesicles. A minority of intercalated cells exhibited basolateral plasma membrane label, and only an occasional cell displayed apical plasma membrane label. In acid-loaded rabbits, H(+)-ATPase immunoreactivity was enhanced along the apical plasma membrane of intercalated cells in the inner CCD, and morphometric analysis demonstrated increased apical plasma membrane in band 3-positive intercalated cells in this segment. These results suggest that rabbits respond to acid loading via enhancement of both electrogenic proton secretion and Cl-/HCO3- exchange in intercalated cells in the inner CCD and the OMCDo.

The "coat" of kidney intercalated cell tubulovesicles does not contain clathrin

1986 ◽  
Vol 250 (4) ◽  
pp. C605-C608 ◽  
Author(s):  
D. Brown ◽  
L. Orci
Keyword(s):  
Coat Protein ◽  
Rat Kidney ◽  
Cell Types ◽  
Intercalated Cells ◽  
Coated Pits ◽  
Thin Sections ◽  
Intercalated Cell ◽  
Apical Cytoplasm ◽  
Collecting Ducts ◽  
Lowicryl K4m

Intercalated cells of kidney collecting ducts contain a population of tubulovesicles in their apical cytoplasm, whose limiting membranes are decorated by arrays of dense, club-shaped projections oriented toward the cytoplasm. These tubulovesicles have been implicated in endo-exocytotic events in these cells. To determine a possible relationship between this “coating” material and clathrin, the coat protein associated with endocytotic coated pits and coated vesicles in other cell types, we applied a monospecific, affinity-purified anti-clathrin antibody to thin sections of rat kidney embedded at low temperature in Lowicryl K4M. We found that no specific labeling was associated with the studlike material of intercalated cell tubulovesicles.

Membrane recycling and epithelial cell function

1989 ◽  
Vol 256 (1) ◽  
pp. F1-F12 ◽  
Author(s):  
D. Brown
Keyword(s):  
Plasma Membrane ◽  
Cell Function ◽  
Collecting Duct ◽  
Water Channel ◽  
Cell Types ◽  
Apical Plasma Membrane ◽  
Proton Pumps ◽  
Intercalated Cells ◽  
Membrane Recycling ◽  
Membrane Composition

The plasma membrane composition of virtually all eucaryotic cells is established, maintained, and modified by the process of membrane recycling. Specific plasma membrane components are inserted by exocytosis of transport vesicles, and are removed by endocytosis of segments of the membrane in which particular proteins are concentrated. In the kidney collecting duct, vasopressin induces the cycling of vesicles that are thought to carry water channels to and from the apical plasma membrane of principal cells, thus modulating the water permeability of this membrane. In the intercalated cells of the collecting duct, hydrogen ion secretion is controlled by the recycling of vesicles carrying proton pumps to and from the plasma membrane. In both cell types, "coated" carrier vesicles are involved, but whereas clathrin-coated vesicles participate in water channel recycling, the vesicles in intercalated cells are coated with the cytoplasmic domains of proton pumps. Following a brief outline of membrane recycling in general, this review summarizes previous data on membrane recycling in the collecting duct and related transporting epithelia and discusses some selected points relating to the role of membrane recycling and cell-specific function in the collecting duct.

Sign in / Sign up

Export Citation Format

Share Document