1. Alterations in vesicle transport and cell polarity in rat hepatocytes subjected to mechanical or chemical cholestasis 
Natalie J. Török, Elizabeth M. Larusso, Mark A. McNiven 
Gastroenterology 
Volume 121, Issue 5, Pages (November 2001)
DOI: /gast
Copyright © 2001 American Gastroenterological Association Terms and Conditions

2. Fig. 1
The number of vesicle movements is markedly reduced in hepatocytes isolated from cholestatic rats. Video computer microscopic analysis of vesicle movements in cultured hepatocytes from control, BDL-treated, and EE-injected rats. The bar graph summarizes the results of the quantitation of vesicular motility. The number of vesicle movements within a defined cytoplasmic region was observed for 5 minutes, recorded, and counted by video microscopy. Vesicles were counted in different regions of 20 different cells per experiment. The experiments were performed 6 times on cells derived from each BDL-treated rat and 3 times on cells from EE-treated rats. Animals that underwent sham operations (BDL) or received injections of only propylene glycol (EE) were used as controls. Control and cholestatic cells were divided into 2 groups: those with few moving vesicles (n = 0-5) and those with many moving vesicles (n = 6-25). In control cells an average of 6-15 vesicles moved in each cytoplasmic region; few or no vesicle movements were observed in BDL- and EE-treated rats.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

3. Fig. 2
Large cytoplasmic lumina in hepatocytes isolated from cholestatic rat livers. Phase video micrographs of cultured hepatocytes isolated from (A, D) control, (B, C) BDL-treated, and (E, F) and EE-treated rats. Although control cells show no abnormal structures, numerous clear, cytoplasmic lumina (arrows) of 1-5 μm diameter are seen throughout the cytoplasm of hepatocytes from cholestatic livers. N, nuclei; bar = 10 μm.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

4. Fig. 3
Quantitation of PCs in cholestatic liver tissues and hepatocytes isolated from cholestatic livers. Quantitation of PCs was performed on EM specimens in control and cholestatic (BDL- and EE-treated) tissues and cells; 200 cells were observed in each case. Comparison of the data shows that only 1.2% and 1.1% of cells in control livers and cells isolated from control livers, respectively, have PCs. Cholestasis induced formation of these structures in 24% of BDL-treated tissues and 32.5% of cells isolated from BDL-treated livers. EE induced formation of PCs in 19% of cells in EE-exposed livers and 10.4% of cells isolated from EE-exposed livers.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

5. Fig. 4
EM of hepatocytes from cholestatic livers shows numerous microvilli within the cytoplasmic lumina. (A, C) Low- and (B, D) higher-magnification EMs of cholestatic hepatocytes. Large cytoplasmic lumina (arrows), 1-5 μm in diameter, are easily distinguished from small empty lipid droplets by the presence of numerous microvilli that project from the limiting luminal membrane. Multiple lipid lamellar bodies (*), normally observed in bile-filled canaliculi, are also observed within the lumina.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

6. Fig. 5
Filamentous actin is arranged around canaliculi and intracellular lumina or PCs in hepatocytes isolated from cholestatic rat livers. (A, C, E) Phase and (B, D, F) corresponding fluorescence rhodamine phalloidin staining of hepatocytes isolated from rats treated with (A-D) EE and (E, F) BDL. Cells are viewed on the day of plating and still have intracellular canaliculi (couplets or triplets). (B, D) Actin staining is restricted about the canaliculi and to large intracellular lumina that appear as PCs (arrows) in the same cells. No connections are seen between the bile canaliculi and the lumina when different planes are viewed. N, nuclei; C, canaliculi; bar = 10 μm.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

7. Fig. 6
Cytoplasmic vesicles containing pIgAR are transported to, and positioned around, PCs. Cultured hepatocytes isolated from BDL rats were stained with a polyclonal antibody to the cytoplasmic tail of pIgAR. In each cell, several PCs are observed (*), surrounded by numerous distinct vesicles containing pIgAR (arrows). In normal hepatocytes cultured from control livers, these vesicles are known to be transported to the pericanalicular region (data not shown). N, nuclei; bar = 10 μm.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

8. Fig. 7
Canaliculi and PCs actively sequester sodium fluorescein. (A, C, E) Phase and (B, D, F) corresponding fluorescence micrographs of a polarized hepatocyte couplet isolated from (A, B) a control rat and (C, E) rats subjected to BDL. (A, B) In control cells, sodium fluorescein was actively sequestered into the canalicular lumen, as has been shown previously by others. In hepatocytes isolated from cholestatic rats, sodium fluorescein was sequestered into the PCs (arrows). N, nuclei; bc, bile canaliculus; bar = 10 μm.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions

9. Fig. 8
EM of cholestatic (BDL- and EE-treated) livers shows cytoplasmic lumina. (A, B) Low- and (C, D) higher-magnification EMs of cholestatic liver tissues. Livers were perfused with paraformaldehyde then processed for EM, as described in Materials and Methods. Large cytoplasmic lumina (arrows) are easily observed in both experimental conditions. The PCs are filled with circular lipid-containing structures consistent with bile accumulations found in conventional canaliculi. (A) By serial sectioning, no connection was found between the bile canaliculi and the lumina.
Gastroenterology  , DOI: ( /gast )
Copyright © 2001 American Gastroenterological Association Terms and Conditions