A basolateral CHIP28/MIP26-related protein (BLIP) in kidney principal cells and gastric parietal cells

1994 ◽  
Vol 267 (3) ◽  
pp. C812-C820 ◽  
Author(s):  
G. Valenti ◽  
J. M. Verbavatz ◽  
I. Sabolic ◽  
D. A. Ausiello ◽  
A. S. Verkman ◽  
...  
Keyword(s):  
Collecting Duct ◽  
Basolateral Membrane ◽  
Water Channel ◽  
Protein S ◽  
High Water ◽  
Protein Band ◽  
Parietal Cells ◽  
Toad Urinary Bladder ◽  
Principal Cells ◽  
Gastric Parietal Cells

The water channel CHIP28 accounts for the high water permeability of proximal tubules and thin descending limbs of Henle; a homologous water channel, WCH-CD, in the apical membrane of collecting duct principal cells, may be the vasopressin-sensitive water channel. We show here that one antiserum, raised against CHIP28, immunostains the basolateral membrane of collecting duct principal cells, in addition to staining CHIP28 in other cells. This serum was named anti-basolateral integral protein (anti-BLIP) to distinguish it from other anti-CHIP28 antisera. By Western blotting, BLIP serum recognized both CHIP28 and MIP26, and it stained lens fibers, which contain MIP26 but not CHIP28. BLIP antiserum immunoprecipitated a 28-kDa band, a broad 35- to 50-kDa band, and an approximately 16-kDa band from kidney papilla. It also stained the basolateral membrane of gastric parietal cells, which were not stained with anti-CHIP28 or anti-MIP26 antibodies. BLIP antiserum immunoprecipitated a 28-kDa protein band from stomach; this protein was not precipitated by anti-CHIP28 antibodies. These results suggest that basolateral membranes of principal cells and parietal cells contain a protein(s) that shares common epitopes with CHIP28 and MIP26. Finally, BLIP but not CHIP28 antiserum stained mesothelial (but not epithelial) cells of toad urinary bladder, a further indication that the BLIP antiserum recognizes a protein distinct from CHIP28.

Localization of MIWC and GLIP water channel homologs in neuromuscular, epithelial and glandular tissues

1995 ◽  
Vol 108 (9) ◽  
pp. 2993-3002 ◽  
Author(s):  
A. Frigeri ◽  
M.A. Gropper ◽  
F. Umenishi ◽  
M. Kawashima ◽  
D. Brown ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Urinary Bladder ◽  
Collecting Duct ◽  
Basolateral Membrane ◽  
Water Channel ◽  
Parietal Cells ◽  
Immunogold Electron Microscopy ◽  
Transitional Epithelium ◽  
Principal Cells ◽  
Kidney Collecting Duct

It was shown recently that water channel homologs MIWC (mercurial insensitive water channel) and GLIP (glycerol intrinsic protein) colocalized in basolateral membranes of kidney collecting duct, tracheal and colonic epithelia, and in brain pia mater. We report here an extensive immunolocalization study of MIWC and GLIP in non-epithelial and glandular epithelial tissues in rat. Immunogold electron microscopy confirmed colocalization of MIWC and GLIP in basolateral membrane of principal cells in kidney collecting duct. However, in other epithelia, MIWC but not GLIP was expressed in basolateral membrane of parietal cells in stomach, and in excretory tubules of salivary and lacrimal glands; GLIP but not MIWC was expressed in transitional epithelium of urinary bladder and skin epidermis. In the central nervous system, MIWC was strongly expressed in the ependymal layer lining the aqueductal system, and in astrocytes throughout the spinal cord and in selected regions of brain. MIWC was also expressed in a plasma membrane pattern in skeletal, but not smooth or cardiac muscle. Neither protein was expressed in small intestine, testis, liver, spleen and nerve. The tissue-specific expression of MIWC suggests a role in fluid transport and/or cell volume regulation in stomach and glandular epithelia. The functional role of MIWC expression in the neuromuscular system and of GLIP expression in skin and urinary bladder is uncertain. The specific cellular sites of MIWC expression (astrocytes, trachea, sarcolemma, gastric parietal cells and kidney principal cells) correspond exactly to sites where orthogonal arrays of particles (OAPs) have been visualized by freeze-fracture electron microscopy, suggesting that MIWC may be the OAP protein.

Immunolocalization and effect of dehydration on AQP3, a basolateral water channel of kidney collecting ducts

1997 ◽  
Vol 272 (2) ◽  
pp. F235-F241 ◽  
Author(s):  
K. Ishibashi ◽  
S. Sasaki ◽  
K. Fushimi ◽  
T. Yamamoto ◽  
M. Kuwahara ◽  
...  
Keyword(s):  
Xenopus Oocytes ◽  
Collecting Duct ◽  
Basolateral Membrane ◽  
Broad Band ◽  
Water Channel ◽  
In Vitro Translation ◽  
Mercury Chloride ◽  
Channel Activity ◽  
Kidney Medulla ◽  
Principal Cells

Aquaporin-3 (AQP3) is unique in its structure (lowest homology with other aquaporins) and in its function (significantly conductive to both small nonelectrolytes and water). However, there is a controversy among researchers on its water transport and induction by dehydration. We examined its localization and the effect of dehydration on its expression in the kidney, as well as its water channel activity when expressed in Xenopus oocytes. In vitro translation using reticulocyte lysate revealed that the size of rat AQP3 was 26 kDa, and the band shifted to around 31 kDa with microsomal fraction, which was sensitive to the digestion with N-glycosidase F. In Western blot analysis of rat kidney medulla, AQP3 appeared as a sharp band at 27 kDa and a broad band at 34-40 kDa. In immunohistochemistry, AQP3 was localized to principal cells and absent in intercalated cells in outer medulla. In inner medulla, AQP3 was restricted to inner medullary collecting duct (IMCD) cells. AQP3 was confined to the basolateral membrane of these cells. Although dehydration of rats for 2 days did not change the distribution pattern of AQP3 in IMCD cells, the dehydration increased AQP3 mRNA by twofold with slight increase of its protein level in kidney medulla. Finally, we confirmed its water channel activity when expressed in Xenopus oocytes. The human AQP3 stimulated osmotic water permeability by eightfold, which was inhibited by 0.3 mM mercury chloride by 34% and reversed by beta-mercaptoethanol. Our results indicate that AQP3 is a glycosylated protein and a mercury-sensitive water channel localized at the basolateral membrane of principal cells and IMCD cells, and its expression is induced by dehydration at both protein and mRNA level.

Vasopressin increases AQP-CD water channel in apical membrane of collecting duct cells in Brattleboro rats

1995 ◽  
Vol 268 (6) ◽  
pp. C1546-C1551 ◽  
Author(s):  
T. Yamamoto ◽  
S. Sasaki ◽  
K. Fushimi ◽  
K. Ishibashi ◽  
E. Yaoita ◽  
...  
Keyword(s):  
Water Permeability ◽  
Apical Membrane ◽  
Collecting Duct ◽  
Basolateral Membrane ◽  
Water Channel ◽  
High Water ◽  
Brattleboro Rats ◽  
Duct Cells ◽  
Cytoplasmic Vesicles ◽  
Collecting Duct Cells

The effect of vasopressin on subcellular localization of AQP-CD and AQP3 water channels was examined in thirsted Brattleboro rats by immunohistochemistry and immunoelectron microscopy. AQP-CD was mainly present in the cytoplasm of the collecting duct cells in association with cytoplasmic vesicles but was sparse in the apical membrane in control vehicle-injected rats. In rats given vasopressin 15 min before death, the number of immunogold particles for AQP-CD in the apical membrane increased significantly (P < 0.002) from 1.8 +/- 0.2 to 10.0 +/- 0.4/microns with a significant decrease (P < 0.05) of cytoplasmic labeling from 32.6 +/- 6.4 to 24.6 +/- 5.6/microns 2, indicating that AQP-CD is the vasopressin-regulated water channel predicted by the “shuttle” hypothesis. In contrast, AQP3 was restricted to the basolateral membrane of the collecting duct cells, and the labeling density of AQP3 was unchanged by vasopressin treatment, indicating that AQP3 is constitutively expressed and may maintain high water permeability of the basolateral membrane.

scholarly journals Basolateral targeting and microtubule-dependent transcytosis of the aquaporin-2 water channel

2013 ◽  
Vol 304 (1) ◽  
pp. C38-C48 ◽  
Author(s):  
Naofumi Yui ◽  
Hua A. J. Lu ◽  
Ying Chen ◽  
Naohiro Nomura ◽  
Richard Bouley ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Cold Shock ◽  
Collecting Duct ◽  
Basolateral Membrane ◽  
Water Channel ◽  
Apical Plasma Membrane ◽  
Indirect Pathway ◽  
Basolateral Membranes ◽  
Principal Cells ◽  
Aquaporin 2

The aquaporin-2 (AQP2) water channel relocates mainly to the apical plasma membrane of collecting duct principal cells after vasopressin (VP) stimulation. AQP2 transport to this membrane domain is assumed to be a direct route involving recycling of intracellular vesicles. However, basolateral plasma membrane expression of AQP2 is observed in vivo in principal cells. Here, we asked whether there is a transcytotic pathway of AQP2 trafficking between apical and basolateral membranes. We used MDCK cells in which AQP2 normally accumulates apically after VP exposure. In contrast, both site-specific biotinylation and immunofluorescence showed that AQP2 is strongly accumulated in the basolateral membrane, along with the endocytic protein clathrin, after a brief cold shock (4°C). This suggests that AQP2 may be constitutively targeted to basolateral membranes and then retrieved by clathrin-mediated endocytosis at physiological temperatures. Rab11 does not accumulate in basolateral membranes after cold shock, suggesting that the AQP2 in this location is not associated with Rab11-positive vesicles. After rewarming (37°C), basolateral AQP2 staining is diminished and it subsequently accumulates at the apical membrane in the presence of VP/forskolin, suggesting that transcytosis can be followed by apical insertion of AQP2. This process is inhibited by treatment with colchicine. Our data suggest that the cold shock procedure reveals the presence of microtubule-dependent AQP2 transcytosis, which represents an indirect pathway of apical AQP2 delivery in these cells. Furthermore, our data indicate that protein polarity data obtained from biotinylation assays, which require cells to be cooled to 4°C during the labeling procedure, should be interpreted with caution.

scholarly journals Polarized Distribution of IQGAP Proteins in Gastric Parietal Cells and Their Roles in Regulated Epithelial Cell Secretion

2003 ◽  
Vol 14 (3) ◽  
pp. 1097-1108 ◽  
Author(s):  
Rihong Zhou ◽  
Zhen Guo ◽  
Charles Watson ◽  
Emily Chen ◽  
Rong Kong ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Actin Cytoskeleton ◽  
Acid Secretion ◽  
Rho Gtpase ◽  
Apical Membrane ◽  
Basolateral Membrane ◽  
Parietal Cells ◽  
Actin Dynamics ◽  
Cell Secretion ◽  
Gastric Parietal Cells

Actin cytoskeleton plays an important role in the establishment of epithelial cell polarity. Cdc42, a member of Rho GTPase family, modulates actin dynamics via its regulators, such as IQGAP proteins. Gastric parietal cells are polarized epithelial cells in which regulated acid secretion occurs in the apical membrane upon stimulation. We have previously shown that actin isoforms are polarized to different membrane domains and that the integrity of the actin cytoskeleton is essential for acid secretion. Herein, we show that Cdc42 is preferentially distributed to the apical membrane of gastric parietal cells. In addition, we revealed that two Cdc42 regulators, IQGAP1 and IQGAP2, are present in gastric parietal cells. Interestingly, IQGAP2 is polarized to the apical membrane of the parietal cells, whereas IQGAP1 is mainly distributed to the basolateral membrane. An IQGAP peptide that competes with full-length IQGAP proteins for Cdc42-binding in vitro also inhibits acid secretion in streptolysin-O-permeabilized gastric glands. Furthermore, this peptide disrupts the association of IQGAP and Cdc42 with the apical actin cytoskeleton and prevents the apical membrane remodeling upon stimulation. We propose that IQGAP2 forms a link that associates Cdc42 with the apical cytoskeleton and thus allows for activation of polarized secretion in gastric parietal cells.

The vasopressin-regulated urea transporter in renal inner medullary collecting duct

1990 ◽  
Vol 259 (3) ◽  
pp. F393-F401 ◽  
Author(s):  
M. A. Knepper ◽  
R. A. Star
Keyword(s):  
Apical Membrane ◽  
Collecting Duct ◽  
Water Channel ◽  
Toad Urinary Bladder ◽  
Urea Transport ◽  
Urea Transporter ◽  
Transport Pathway ◽  
Inner Medullary Collecting Duct ◽  
Urea Permeability ◽  
Medullary Collecting Duct

The terminal part of the inner medullary collecting duct (terminal IMCD) is unique among collecting duct segments in part because its permeability to urea is regulated by vasopressin. The urea permeability can rise to extremely high levels (greater than 100 x 10(-5) cm/s) in response to vasopressin. Recent studies in isolated perfused IMCD segments have established that the rapid movement of urea across the tubule epithelium occurs via a specialized urea transporter, presumably an intrinsic membrane protein, present in both the apical and basolateral membranes. This urea transporter has properties similar to those of the urea transporters in mammalian erythrocytes and in toad urinary bladder, namely, inhibition by phloretin, inhibition by urea analogues, saturation kinetics in equilibrium-exchange experiments, and regulation by vasopressin. The urea transport pathway is distinct from and independent of the vasopressin-regulated water channel. The increase in transepithelial urea transport in response to vasopressin is mediated by adenosine 3',5'-cyclic monophosphate and is associated with an increase in the urea permeability of the apical membrane. However, little is known about the physical events associated with the activation or insertion of urea transporters in the apical membrane. Because of the importance of this transporter to the urinary concentrating mechanism, efforts toward understanding its molecular structure and the molecular basis of its regulation appear to be justified.

Depletion of intercalated cells from collecting ducts of carbonic anhydrase II-deficient (CAR2 null) mice

1995 ◽  
Vol 269 (6) ◽  
pp. F761-F774 ◽  
Author(s):  
S. Breton ◽  
S. L. Alper ◽  
S. L. Gluck ◽  
W. S. Sly ◽  
J. E. Barker ◽  
...  
Keyword(s):  
Carbonic Anhydrase ◽  
Collecting Duct ◽  
Water Channel ◽  
Immunofluorescent Staining ◽  
Specific Marker ◽  
Intercalated Cells ◽  
Cortical Collecting Duct ◽  
Principal Cells ◽  
Collecting Tubules ◽  
Deficient Mice

The kidneys of mice (CAR2-null mice) that are genetically devoid of carbonic anhydrase type II (CAII) were screened by immunocytochemistry with antibodies that distinguish intercalated and principal cells. Immunofluorescent localization of the anion exchanger AE1 and of the 56-kDa subunit of the vacuolar H(+)-adenosinetriphosphatase (H(+)-ATPase) was used to identify intercalated cells, while the AQP2 water channel was used as a specific marker for principal cells of the collecting duct. The CAII deficiency of the CAR2-null mice was first confirmed by the absence of immunofluorescent staining of kidney sections exposed to an anti-CAII antibody. Cells positive for AE1 and H(+)-ATPase were common in all collecting duct regions in normal mice but were virtually absent from the inner stripe of the outer medulla and the inner medulla of CAR2-null mice. The number of positive cells was also reduced threefold in the cortical collecting duct of CAR2-null animals compared with normal mice. In parallel, the percentage of AQP2-positive cells was correspondingly increased in the collecting tubules of CAII-deficient mice, whereas the total number of cells per tubule remained unchanged. These results suggest that intercalated cells are severely depleted and are replaced by principal cells in CAII-deficient mice. Quantitative analysis and double staining showed that, in the cortex, both type A and type B intercalated cells are equally affected. Elucidation of the mechanism(s) responsible for this phenotype will be of importance in understanding the origin and development of intercalated cells in the kidney.

Structural-functional relationships along the distal nephron

1986 ◽  
Vol 250 (1) ◽  
pp. F1-F15 ◽  
Author(s):  
K. M. Madsen ◽  
C. C. Tisher
Keyword(s):  
Atpase Activity ◽  
Collecting Duct ◽  
Basolateral Membrane ◽  
Distal Tubule ◽  
Structural Heterogeneity ◽  
Apical Plasma Membrane ◽  
Thick Ascending Limb ◽  
Intercalated Cells ◽  
Principal Cells ◽  
Functional Relationships

The distal tubule, which includes the thick ascending limb (TAL), the macula densa, and the distal convoluted tubule (DCT), and the collecting duct are structurally heterogeneous, thus reflecting the functional heterogeneity that is also present. As the TAL ascends from medulla to cortex, the surface area of the apical plasma membrane increases while that of the basolateral membrane decreases. The structure of the DCT resembles that of the medullary TAL. An excellent correlation exists between structure, Na-K-ATPase activity, and NaCl reabsorptive capacity in the distal tubule. The collecting duct is subdivided into the initial collecting tubule (ICT), and cortical (CCD), outer medullary (OMCD), and inner medullary (IMCD) collecting ducts. Between the distal tubule and the collecting duct is a transition region termed the connecting segment or connecting tubule (CNT). Considerable structural heterogeneity exists along the collecting duct within the two major cell populations, the intercalated cells and the principal cells. In the CNT, the ICT, and the CCD, potassium loading and mineralocorticoids stimulate Na-K-ATPase activity and cause proliferation of the basolateral membrane of CNT cells and principal cells, thus identifying the cells responsible for mineralocorticoid-stimulated potassium secretion in these regions. Finally, at least two morphologically distinct populations of intercalated cells exist, types A and B. In the rat, type A predominates in the CNT and the OMCD and is believed to be responsible for H+ secretion, at least in the OMCD. Type B predominates in the CCD, where it may be involved in bicarbonate secretion.

scholarly journals Insulin and IGF-1 activate Kir4.1/5.1 channels in cortical collecting duct principal cells to control basolateral membrane voltage

2016 ◽  
Vol 310 (4) ◽  
pp. F311-F321 ◽  
Author(s):  
Oleg Zaika ◽  
Oleg Palygin ◽  
Viktor Tomilin ◽  
Mykola Mamenko ◽  
Alexander Staruschenko ◽  
...  
Keyword(s):  
Single Channel ◽  
Collecting Duct ◽  
Basolateral Membrane ◽  
Sodium Reabsorption ◽  
Cortical Collecting Duct ◽  
Open Probability ◽  
Whole Cell ◽  
Principal Cells ◽  
Basolateral Side ◽  
Kinase Cascade

Potassium Kir4.1/5.1 channels are abundantly expressed at the basolateral membrane of principal cells in the cortical collecting duct (CCD), where they are thought to modulate transport rates by controlling transepithelial voltage. Insulin and insulin-like growth factor-1 (IGF-1) stimulate apically localized epithelial sodium channels (ENaC) to augment sodium reabsorption in the CCD. However, little is known about their actions on potassium channels localized at the basolateral membrane. In this study, we implemented patch-clamp analysis in freshly isolated murine CCD to assess the effect of these hormones on Kir4.1/5.1 at both single channel and cellular levels. We demonstrated that K+-selective conductance via Kir4.1/5.1 is the major contributor to the macroscopic current recorded from the basolateral side in principal cells. Acute treatment with 10 μM amiloride (ENaC blocker), 100 nM tertiapin-Q (TPNQ; ROMK inhibitor), and 100 μM ouabain (Na+-K+-ATPase blocker) failed to produce a measurable effect on the macroscopic current. In contrast, Kir4.1 inhibitor nortriptyline (100 μM), but not fluoxetine (100 μM), virtually abolished whole cell K+-selective conductance. Insulin (100 nM) markedly increased the open probability of Kir4.1/5.1 and nortriptyline-sensitive whole cell current, leading to significant hyperpolarization of the basolateral membrane. Inhibition of the phosphatidylinositol 3-kinase cascade with LY294002 (20 μM) abolished action of insulin on Kir4.1/5.1. IGF-1 had similar stimulatory actions on Kir4.1/5.1-mediated conductance only when applied at a higher (500 nM) concentration and was ineffective at 100 nM. We concluded that both insulin and, to a lesser extent, IGF-1 activate Kir4.1/5.1 channel activity and open probability to hyperpolarize the basolateral membrane, thereby facilitating Na+ reabsorption in the CCD.

scholarly journals Prostaglandin E2-Activated Housekeeping Cl- Channels in the Basolateral Membrane of Rat Gastric Parietal Cells.

1999 ◽  
Vol 49 (4) ◽  
pp. 365-372 ◽  
Author(s):  
Akira IKARI ◽  
Hideki SAKAI ◽  
Akiko TANAKA ◽  
Atsushi IKEDA ◽  
Kanako INOUE ◽  
...  
Keyword(s):  
Prostaglandin E2 ◽  
Basolateral Membrane ◽  
Parietal Cells ◽  
Cl Channels ◽  
Gastric Parietal Cells
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