Constitutive and inducible stress proteins dominate the proteome of the murine inner medullary collecting duct-3 (mIMCD3) cell line

2006 ◽  
Vol 1764 (6) ◽  
pp. 1007-1020 ◽  
Author(s):  
Nelly Valkova ◽  
Dietmar Kültz
Keyword(s):  
Cell Line ◽  
Stress Proteins ◽  
Collecting Duct ◽  
Inner Medullary Collecting Duct ◽  
Medullary Collecting Duct

scholarly journals Characterization of anion exchangers in an inner medullary collecting duct cell line.

1998 ◽  
Vol 9 (5) ◽  
pp. 746-754
Author(s):  
G Obrador ◽  
H Yuan ◽  
T M Shih ◽  
Y H Wang ◽  
M A Shia ◽  
...  
Keyword(s):  
Cell Line ◽  
Anion Exchanger ◽  
Collecting Duct ◽  
Basolateral Membrane ◽  
Disulfonic Acid ◽  
Kidney Cortex ◽  
Intercalated Cells ◽  
Rat Stomach ◽  
Inner Medullary Collecting Duct ◽  
Medullary Collecting Duct

Although the inner medullary collecting duct (IMCD) plays a major role in urinary acidification, the molecular identification of many of the specific components of the transport system in this nephron segment are lacking. A cultured line of rat IMCD cells was used to characterize the mediators of cellular HCO3 exit. This cell line functionally resembles alpha-intercalated cells. Physiologic experiments document that HCO3- transport is a reversible, electroneutral, Cl dependent, Na+-independent process. It can be driven by Cl-gradients and inhibited by stilbenes such as 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid. Immunohistochemical analysis, using a rabbit polyclonal antibody against the carboxy-terminal 12 amino acids of anion exchanger 1 (AE1), revealed a distribution of immunoreactive protein that is consistent with a basolateral localization of AE in cultured cells and in alpha-intercalated cells identified in sections of rat kidney cortex. Immunoblot revealed two immunoreactive bands (approximately 100 and 180 kD in size) in membranes from cultured IMCD cells, rat renal medulla, and freshly isolated IMCD cells. The mobility of the lower molecular weight band was similar to that of AE1 in red blood cell ghosts and kidney homogenate and therefore probably represents AE1. The mobility of the 180-kD band is similar to that for rat stomach and kidney AE2 and therefore probably represents AE2. Selective biotinylation of the apical or basolateral membrane proteins in cultured IMCD cells revealed that both AE1 and AE2 are polarized to the basolateral membrane. Northern blot analysis documented the expression of mRNA for AE1 and AE2 but not AE3. Furthermore, the cDNA sequence of AE1 and AE2 expressed by these cells was found to be virtually identical to that reported for kidney AE1 and rat stomach AE2. It is concluded that this cultured line of rat IMCD cells expresses two members of the anion exchanger gene family, AE1 and AE2, and both of these exchangers probably mediate the electroneutral Cl--dependent HCO3-transport observed in this cell line.

Early transcriptional effects of aldosterone in a mouse inner medullary collecting duct cell line

2003 ◽  
Vol 285 (4) ◽  
pp. F664-F673 ◽  
Author(s):  
Michelle L. Gumz ◽  
Michael P. Popp ◽  
Charles S. Wingo ◽  
Brian D. Cain
Keyword(s):  
Cell Line ◽  
Cellular Response ◽  
Collecting Duct ◽  
Duct Cell ◽  
Immunoblot Analysis ◽  
Tissue Growth ◽  
Long Term Effects ◽  
Acid Base Balance ◽  
Inner Medullary Collecting Duct ◽  
Medullary Collecting Duct

The mineralocorticoid aldosterone is a major regulator of Na+ and acid-base balance and control of blood pressure. Although the long-term effects of aldosterone have been extensively studied, the early aldosterone-responsive genes remain largely unknown. Using DNA array technology, we have characterized changes in gene expression after 1 h of exposure to aldosterone in a mouse inner medullary collecting duct cell line, mIMCD-3. Results from three independent microarray experiments revealed that the expression of many transcripts was affected by aldosterone treatment. Northern blot analysis confirmed the upregulation of four distinct transcripts identified by the microarray analysis, namely, the serum and glucose-regulated kinase sgk, connective tissue growth factor, period homolog, and preproendothelin. Immunoblot analysis for preproendothelin demonstrated increased protein expression. Following the levels of the four transcripts over time showed that each had a unique pattern of expression, suggesting that the cellular response to aldosterone is complex. The results presented here represent a novel list of early aldosterone-responsive transcripts and provide new avenues for elucidating the mechanism of acute aldosterone action in the kidney.

Electrogenic sodium absorption and chloride secretion by an inner medullary collecting duct cell line (mIMCD-K2)

1995 ◽  
Vol 268 (2) ◽  
pp. F347-F355 ◽  
Author(s):  
N. L. Kizer ◽  
B. Lewis ◽  
B. A. Stanton
Keyword(s):  
Cell Line ◽  
Collecting Duct ◽  
T Antigen ◽  
Sodium Absorption ◽  
Sv40 Large T Antigen ◽  
Permanent Cell Line ◽  
Inner Medullary Collecting Duct ◽  
Permanent Cell ◽  
Medullary Collecting Duct

The initial segment of the inner medullary collecting duct (IMCDi) absorbs Na+ by an electrogenic mechanism and plays an important role in regulating the composition and volume of the urine. The purpose of the present study was to establish a permanent cell line derived from the IMCDi, which has the ion transport properties of the IMCDi in vivo. To this end, we isolated IMCD cells from the IMCDi of a mouse, Tg(SV40E) Bri 7, transgenic for the early region of SV40 (large T antigen) and established a permanent cell line, mIMCD-K2, by clonal dilution. mIMCD-K2 cells retain many differentiated characteristics of the IMCDi, including amiloride-sensitive electrogenic Na+ absorption stimulated by nanomolar concentrations of aldosterone. Aldosterone (1.5 x 10(-6) M) increased Na+ absorption from 0.2 +/- 0.1 to 4.6 +/- 1.7 microA/cm2. In addition, the cells secrete Cl- by an electrogenic mechanism at a rate of 0.5 +/- 0.1 microA/cm2. We propose that IMCDi cells either absorb or secrete NaCl depending on NaCl homeostasis. The mIMCD-K2 cell line should be useful for studying the cellular mechanisms responsible for electrogenic Na+ and Cl- transport in the IMCDi.

H+ secretion is inhibited by clostridial toxins in an inner medullary collecting duct cell line

1997 ◽  
Vol 273 (6) ◽  
pp. F1054-F1057 ◽  
Author(s):  
Edward A. Alexander ◽  
Theodora Shih ◽  
John H. Schwartz
Keyword(s):  
Epithelial Cell ◽  
Cell Line ◽  
Collecting Duct ◽  
Duct Cell ◽  
Specific Protein ◽  
Proton Pumps ◽  
Renal Epithelial Cell ◽  
Inner Medullary Collecting Duct ◽  
Medullary Collecting Duct ◽  
Cell Acid

Renal epithelial cell H+secretion is an exocytic-endocytic phenomenon. In the inner medullary collecting duct (IMCD) cell line, which we have utilized as a model of renal epithelial cell acid secretion, we found previously that acidification increased exocytosis and alkalinization increased endocytosis. It is likely, therefore, that the rate of proton secretion is regulated by the membrane insertion and retrieval of proton pumps. There is abundant evidence from studies in the nerve terminal and the chromaffin cell that vesicle docking, membrane fusion, and discharge of vesicular contents (exocytosis) involve a series of interactions among so-called trafficking proteins. The clostridial toxins, botulinum and tetanus, are proteases that specifically inactivate some of these proteins. In these experiments we demonstrated, by immunoblot and immunoprecipitation, the presence in this IMCD cell line of the specific protein targets of these toxins, synaptobrevin/vesicle-associated membrane proteins (VAMP), syntaxin, and synaptosomal-associated protein-25 (SNAP-25). Furthermore, we showed that these toxins markedly inhibit the capacity of these cells to realkalinize after an acid load. Thus these data provide new insight into the mechanism for H+ secretion in the IMCD.

An osmotically tolerant inner medullary collecting duct cell line from an SV40 transgenic mouse

1993 ◽  
Vol 265 (3) ◽  
pp. F416-F424 ◽  
Author(s):  
M. I. Rauchman ◽  
S. K. Nigam ◽  
E. Delpire ◽  
S. R. Gullans
Keyword(s):  
Cell Line ◽  
Cell Biology ◽  
Collecting Duct ◽  
T Antigen ◽  
Organic Osmolytes ◽  
Sv40 Large T Antigen ◽  
Hypertonic Medium ◽  
Inner Medullary Collecting Duct ◽  
Nephron Segment ◽  
Medullary Collecting Duct

The terminal inner medullary collecting duct (IMCD) plays an important role in determining the final urinary composition. Currently, there is no continuous cell line derived from this nephron segment. We have developed a cell line derived from the terminal IMCD of mice transgenic for the early region of simian virus SV40 (large T antigen). This cell line, mIMCD-3, retains many differentiated characteristics of this nephron segment including high transepithelial resistance (1,368 +/- 172 omega.cm2), inhibition of apical-to-basal sodium flux by amiloride (41 +/- 7%) and by atrial natriuretic peptide (ANP) (40 +/- 9%), the presence of the amiloride-sensitive sodium channel as determined by Western blot analysis, and accumulation of the major organic osmolytes in response to hypertonic stress. Significantly, mIMCD-3 cells adapted readily and were able to grow in hypertonic medium supplemented with NaCl and urea up to 910 mosmol/kgH2O. These extreme osmotic conditions exist in the renal medulla in vivo but are known to be lethal to most other cells. This cell line should be highly useful for the study of the cellular adaptation to osmotic stress and the cell biology and transport physiology of this nephron segment.

scholarly journals microRNA Regulation of endothelin-1 in an inner medullary collecting duct cell line

Life Sciences ◽  
2013 ◽  
Vol 93 (25-26) ◽  
pp. e37
Author(s):  
Mollie E./INS; Jacobs ◽  
Lauren A./INS; Jeffers ◽  
Amanda K./INS; Welch ◽  
Charles S./INS; Wingo ◽  
Brian D./INS; Cain
Keyword(s):  
Cell Line ◽  
Collecting Duct ◽  
Duct Cell ◽  
Microrna Regulation ◽  
Endothelin 1 ◽  
Inner Medullary Collecting Duct ◽  
Medullary Collecting Duct

Effect of acidification on the location of H+-ATPase in cultured inner medullary collecting duct cells

1999 ◽  
Vol 276 (3) ◽  
pp. C758-C763 ◽  
Author(s):  
Edward A. Alexander ◽  
Dennis Brown ◽  
Theodora Shih ◽  
Mary McKee ◽  
John H. Schwartz
Keyword(s):  
Cell Line ◽  
Apical Membrane ◽  
Collecting Duct ◽  
Renal Epithelial Cell ◽  
Inner Medullary Collecting Duct ◽  
Medullary Collecting Duct ◽  
Cell Acid ◽  
Immunohistochemical Techniques ◽  
Cellular Acidification

In previous studies, our laboratory has utilized a cell line derived from the rat inner medullary collecting duct (IMCD) as a model system for mammalian renal epithelial cell acid secretion. We have provided evidence, from a physiological perspective, that acute cellular acidification stimulates apical exocytosis and elicits a rapid increase in proton secretion that is mediated by an H+-ATPase. The purpose of these experiments was to examine the effect of acute cellular acidification on the distribution of the vacuolar H+-ATPase in IMCD cells in vitro. We utilized the 31-kDa subunit of the H+-ATPase as a marker of the complete enzyme. The distribution of this subunit of the H+-ATPase was evaluated by immunohistochemical techniques (confocal and electron microscopy), and we found that there is a redistribution of these pumps from vesicles to the apical membrane. Immunoblot evaluation of isolated apical membrane revealed a 237 ± 34% ( P < 0.05, n = 9) increase in the 31-kDa subunit present in the membrane fraction 20 min after the induction of cellular acidification. Thus our results demonstrate the presence of this pump subunit in the IMCD cell line in vitro and that cell acidification regulates the shuttling of cytosolic vesicles containing the 31-kDa subunit into the apical membrane.

Sign in / Sign up

Export Citation Format

Share Document