scholarly journals Regulation of vesicular pH in liver macrophages and parenchymal cells by ammonia and anisotonicity as assessed by fluorescein isothiocyanate-dextran fluorescence

1996 ◽  
Vol 315 (2) ◽  
pp. 385-392 ◽  
Author(s):  
Rainer SCHREIBER ◽  
Fan ZHANG ◽  
Dieter HÄUSSINGER
Keyword(s):  
Kupffer Cells ◽  
Ph Value ◽  
Fluorescein Isothiocyanate ◽  
Rat Hepatocytes ◽  
Cell Swelling ◽  
Membrane Flow ◽  
Liver Parenchymal ◽  
Concanamycin A ◽  
Acidic Compartment ◽  
Parenchymal Cells

Short-term-cultivated rat hepatocytes and Kupffer cells were allowed to endocytose fluorescein isothiocyanate (FITC)-coupled dextran, in order to study the effects of aniso-osmotic exposure and NH4Cl on apparent vesicular pH (pHves) by single-cell fluorescence. Following a 2 h loading period with FITC–dextran in normo-osmotic (305 mosmol/l) medium, the apparent pHves was 6.01±0.05 (n = 39) in parenchymal cells and 4.94±0.04 (n = 76) in Kupffer cells. Under these conditions pHves in parenchymal cells, but not in Kupffer cells, was sensitive to changes in ambient osmolarity. Inhibition of vacuolar H+-ATPase by concanamycin A did not affect the osmosensitivity of pHves in parenchymal cells. However, the effects of anisotonicity on pHves were largely abolished in the presence of 4,4´-di-isothiocyanato-stilbene-2,2´-disulphonic acid (DIDS) or when extracellular chloride was substituted for gluconate. In neither Kupffer cells, nor liver parenchymal cells did hypo-osmotic cell swelling cause an increase in intracellular Ca2+. With regard to vesicular acidification, the following differences were noted between parenchymal and Kupffer cells. (1) In Kupffer cells endocytosed FITC–dextran reached a strongly acidic compartment with a pH value of approx. 5 within 5 min, whereas it took 4–5 h in parenchymal cells. Modification of pHves by hypo-osmolarity in Kupffer cells was only observed in a short-lived ‘early’ compartment with a pH value of approx. 6. (2) In contrast to pHves in parenchymal cells, pHves in Kupffer cells was very sensitive towards alkalinization by NH4Cl: addition of NH4Cl at 1 or 10 mM increased apparent pHves by 0.80 or 1.46 in Kupffer cells, but only by 0.18 or 0.56 in parenchymal cells. The low ammonia sensitivity of pHves in parenchymal cells was observed not only in the less acidic (pH approx. 6) endocytotic compartment which is reached by FITC–dextran within 2 h, but also in the stronger acidic compartment (pH approx. 5) which is reached after 4–5 h. (3) NH4Cl had no effect on the osmosensitivity of pHves in parenchymal cells, whereas in Kupffer cells pHves became sensitive to anisotonicity when NH4Cl was present. Osmosensitivity of pHves in Kupffer cells under these conditions, however, was not affected by genistein, DIDS or colchicine, whereas these compounds abolished the osmosensitivity of pHves in parenchymal cells. It is suggested that regulation of pHves by cell volume in liver parenchymal cells involves changes of vesicular chloride conductance. In addition, there are marked differences between Kupffer and parenchymal cells with respect to vesicular ammonia permeability and the kinetics of endocytotic membrane flow and acidification.

scholarly journals Cytochemical localization of beta-NADPase in rat hepatocytes and Kupffer cells. Comparison with thiamine pyrophosphatase (TPPase).

1984 ◽  
Vol 32 (5) ◽  
pp. 541-546 ◽  
Author(s):  
S Angermüller ◽  
H D Fahimi
Keyword(s):  
Golgi Apparatus ◽  
Kupffer Cells ◽  
Intracellular Localization ◽  
Rat Hepatocytes ◽  
The Other ◽  
Cytochemical Localization ◽  
Liver Parenchymal ◽  
Thiamine Pyrophosphatase ◽  
The Difference ◽  
Parenchymal Cells

The intracellular localization of beta-NADPase in rat hepatocytes and Kupffer cells has been studied and compared with the pattern of TPPase in these cells. The reaction product for beta-NADPase is present in some but not all hepatocytes in two cisternae on the trans aspect of the Golgi apparatus. It is absent from the trans-most lamella and the GERL of hepatocytes. TPPase, on the other hand, is limited to the first Golgi cisterna on the trans aspect with sprinkles of reaction product in the second lamella. Considering that TPPase is a marker of the trans Golgi lamella and hepatocyte Golgi stacks contain usually 2-4 lamellae, our observations suggest that beta-NADPase is localized in the trans as well as in the intermediate Golgi lamellae of liver parenchymal cells. In Kupffer cells, the reaction product for both beta-NADPase and TPPase was found in some but not in all cells. The enzyme beta-NADPase was localized in the rigid lamella and the tubulovacuolar system of GERL. This pattern differed significantly from that for TPPase, which was found in 2-3 cisternae at the trans aspect of the Golgi complex in Kupffer cells. These observations demonstrate the difference in the localization of beta-NADPase in hepatocytes and Kupffer cells. Such differences should be taken into consideration in studies of Golgi fractions, when phosphatase reactions are used as specific markers of Golgi components.

scholarly journals Low-density-lipoprotein receptors in different rabbit liver cells

1989 ◽  
Vol 261 (2) ◽  
pp. 587-593 ◽  
Author(s):  
M S Nenseter ◽  
O Myklebost ◽  
R Blomhoff ◽  
C A Drevon ◽  
A Nilsson ◽  
...  
Keyword(s):  
Kupffer Cells ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Binding Activity ◽  
Rabbit Liver ◽  
Low Density ◽  
Apo B ◽  
Liver Parenchymal ◽  
Ldl Binding ◽  
Parenchymal Cells

Receptor-dependent uptake mechanisms for low-density lipoprotein (LDL) were studied in rabbit liver parenchymal and non-parenchymal cells. Hybridization studies with a cDNA probe revealed that mRNA for the apo (apolipoprotein) B,E receptor was present in endothelial and Kupffer cells as well as in parenchymal cells. By ligand-blotting experiments we showed that apo B,E-receptor protein was present in both parenchymal and non-parenchymal cells. Studies of binding of homologous LDL in cultured rabbit parenchymal cells suggested that about 63% of the specific LDL binding was mediated via the apo B,E receptor. Approx. 47% of the specific LDL binding was dependent on Ca2+, suggesting that specific Ca2+-dependent as well as Ca2+-independent LDL-binding sites exist in liver parenchymal cells. Methylated LDL bound to the parenchymal cells in a saturable manner. Taken together, our results showed that apo B,E receptors are present in rabbit liver endothelial and Kupffer cells as well as in the parenchymal cells, and that an additional saturable binding activity for LDL may exist on rabbit liver parenchymal cells. This binding activity was not inhibited by EGTA or reductive methylation of lysine residues in apo B. LDL degradation in parenchymal cells was mainly mediated via the apo B,E receptor.

scholarly journals Effects of aniso-osmolarity and hydroperoxides on intracellular pH in isolated rat hepatocytes as assessed by (2′,7′)-bis(carboxyethyl)-5(6)-carboxyfluorescein and fluorescein isothiocyanate-dextran fluorescence

1994 ◽  
Vol 303 (1) ◽  
pp. 113-120 ◽  
Author(s):  
R Schreiber ◽  
B Stoll ◽  
F Lang ◽  
D Häussinger
Keyword(s):  
Fluorescein Isothiocyanate ◽  
Rat Hepatocytes ◽  
Cell Swelling ◽  
Cell Shrinkage ◽  
Isolated Rat Hepatocytes ◽  
Proton Gradients ◽  
Fluorescein Isothiocyanate Dextran ◽  
And Oxidative Stress ◽  
Vesicular Compartment ◽  
Isolated Rat

Freshly isolated rat hepatocytes were plated for 4-6 h and either loaded with (2′,7)-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF) or allowed to endocytose fluorescein isothiocyanate (FITC)-coupled dextran in order to study the effects of aniso-osmotic exposure and oxidative stress on cytosolic (pHcyt) and apparent vesicular pH (pHves) by single-cell fluorescence recordings. In the presence of normo-osmotic (305 mosmol/l) medium pHcyt was 7.23 +/- 0.03 (n = 108), whereas an apparent pH of 6.07 +/- 0.02 (n = 156) was found in the vesicular compartment accessible to endocytosed FITC-dextran. Substitution of 60 mM NaCl against 120 mM raffinose had no effect on pHcyt or apparent pHves, whereas addition of NH4Cl increased both pHcyt and apparent pHves. Hypo-osmotic cell swelling lowered pHcyt, whereas simultaneously apparent pHves increased. These effects were rapidly reversible upon re-institution of normo-osmotic media. Similarly, an increase of apparent pHves was observed when cell swelling was induced by Ba2+, glutamine or histidine. Conversely, hyperosmotic cell shrinkage due to addition of NaCl or raffinose led to a cytosolic alkalinization and a vesicular acidification. Both, H2O2 (0.2 mmol/l) and t-butyl-hydroperoxide (0.2 mmol/l) were without effect on pHcyt, but lowered apparent pHves by about 0.2 pH units. Ba2+ (1 mmol/l) diminished the acidifying effect of the hydroperoxides by about 50%. Pretreatment of the cells with colchicine, but not with lumicolchicine, largely abolished the effects of aniso-osmolarity and hydroperoxides on pHves. The data suggest that hepatocellular hydration affects the proton gradients built up across the membranes of endocytotic FITC-dextran-accessible compartments in a microtubule-dependent way. They further suggest that hydroperoxides induce vesicular acidification in a colchicine- and Ba(2+)-sensitive way. Because hydroperoxides induce Ba(2+)-sensitive cell shrinkage [Hallbrucker, Ritter, Lang, Gerok and Häussinger (1992) Eur. J. Biochem. 211, 449-458], the results are compatible with the view that hydroperoxide-induced cell shrinkage mediates vesicular acidification. It is concluded that modulation of vesicular pH by the hepatocellular hydration state may play a role in triggering some metabolic changes in response to cell swelling/shrinkage.

Nuclear receptor atlas of female mouse liver parenchymal, endothelial, and Kupffer cells

2013 ◽  
Vol 45 (7) ◽  
pp. 268-275 ◽  
Author(s):  
Zhaosha Li ◽  
J. Kar Kruijt ◽  
Ronald J. van der Sluis ◽  
Theo J. C. Van Berkel ◽  
Menno Hoekstra
Keyword(s):  
Expression Pattern ◽  
Nuclear Receptors ◽  
Nuclear Receptor ◽  
Kupffer Cells ◽  
Virgin Female ◽  
Orphan Receptor ◽  
Orphan Nuclear Receptors ◽  
Liver Parenchymal ◽  
Hepatic Expression ◽  
Parenchymal Cells

The liver consists of different cell types that together synchronize crucial roles in liver homeostasis. Since nuclear receptors constitute an important class of drug targets that are involved in a wide variety of physiological processes, we have composed the hepatic cell type-specific expression profile of nuclear receptors to uncover the pharmacological potential of liver-enriched nuclear receptors. Parenchymal liver cells (hepatocytes) and liver endothelial and Kupffer cells were isolated from virgin female C57BL/6 wild-type mice using collagenase perfusion and counterflow centrifugal elutriation. The hepatic expression pattern of 49 nuclear receptors was generated by real-time quantitative PCR using the NUclear Receptor Signaling Atlas (NURSA) program resources. Thirty-six nuclear receptors were expressed in total liver. FXR-α, EAR2, LXR-α, HNF4-α, and CAR were the most abundantly expressed nuclear receptors in liver parenchymal cells. In contrast, NUR77, COUP-TFII, LXR-α/β, FXR-α, and EAR2 were the most highly expressed nuclear receptors in endothelial and Kupffer cells. Interestingly, members of orphan receptor COUP-TF family showed a distinct expression pattern. EAR2 was highly and exclusively expressed in parenchymal cells, while COUP-TFII was moderately and exclusively expressed in endothelial and Kupffer cells. Of interest, the orphan receptor TR4 showed a similar expression pattern as the established lipid sensor PPAR-γ. In conclusion, our study provides the most complete quantitative assessment of the nuclear receptor distribution in liver reported to date. Our gene expression catalog suggests that orphan nuclear receptors such as COUP-TFII, EAR2, and TR4 may be of significant importance as novel targets for pharmaceutical interventions in liver.

scholarly journals Human recombinant apolipoprotein E redirects lipopolysaccharide from Kupffer cells to liver parenchymal cells in rats In vivo.

1997 ◽  
Vol 99 (10) ◽  
pp. 2438-2445 ◽  
Author(s):  
P C Rensen ◽  
M Oosten ◽  
E Bilt ◽  
M Eck ◽  
J Kuiper ◽  
...  
Keyword(s):  
Apolipoprotein E ◽  
Kupffer Cells ◽  
Liver Parenchymal ◽  
Parenchymal Cells

Transport of tyramine in short-term cultured rat liver parenchymal cells, endothelial cells and Kupffer cells

1995 ◽  
Vol 3 (2) ◽  
pp. 105-111
Author(s):  
Z ZHONG ◽  
M PIROTTEWARNIER ◽  
A VUYLSTEKE ◽  
S CONINCK ◽  
R WATTIAUX
Keyword(s):  
Endothelial Cells ◽  
Rat Liver ◽  
Kupffer Cells ◽  
Short Term ◽  
Liver Parenchymal ◽  
Parenchymal Cells

Cholesteryl esters from oxidized low-density lipoproteins arein vivo rapidly hydrolyzed in rat kupffer cells and transported to liver parenchymal cells and bile

Hepatology ◽  
1994 ◽  
Vol 19 (6) ◽  
pp. 1459-1467
Author(s):  
Moniek N. Pieters ◽  
Sebastiaan Esbach ◽  
Donald Schouten ◽  
Adriaan Brouwer ◽  
Dick L. Knook ◽  
...  
Keyword(s):  
Kupffer Cells ◽  
Low Density Lipoproteins ◽  
Low Density ◽  
Cholesteryl Esters ◽  
Liver Parenchymal ◽  
Oxidized Low Density Lipoproteins ◽  
Parenchymal Cells

Direct evidence of production of nitric oxide in the lipopolysaccharide-sensitized rat hepatocytes and kupffer cells

2001 ◽  
Vol 120 (5) ◽  
pp. A356-A356
Author(s):  
T KONO ◽  
J IWAMOTO ◽  
K ISHIKAWA ◽  
Y EBISAWA ◽  
T AOKI ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Kupffer Cells ◽  
Direct Evidence ◽  
Rat Hepatocytes
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