Determinants of biliary secretion in isolated perfused skate liver

1982 ◽  
Vol 242 (4) ◽  
pp. G319-G325 ◽  
Author(s):  
J. S. Reed ◽  
N. D. Smith ◽  
J. L. Boyer
Keyword(s):  
Tight Junctions ◽  
Bile Flow ◽  
Perfusion Pressure ◽  
Freeze Fracture ◽  
Biliary Tree ◽  
Solute Diffusion ◽  
Electron Microscopic ◽  
Bile Formation ◽  
Ionic Lanthanum ◽  
Microscopic Studies

In the isolated perfused liver of the little skate, Raja erinacea, bile flow averaged 5.07 +/- 0.58 (mean +/- SE) microliters.h-1.g liver-1 in 21 experiments at a perfusion pressure of 5.0 cm Ringer compared to 3.79 +/- 0.32 in 38 experiments at 2.5 cm (P less than 0.05). [14C]inulin readily entered skate bile. Bile-to-plasma [14C]inulin ratios corrected for delay in transit time, averaged 0.46 +/- 0.07 at 1 h and rose to 0.74 +/- 0.06 by 4 h, although bile flow remained constant. In experiments in which [14C]inulin reached equilibrium between bile and plasma, the bile-to-plasma ratio conformed to the theoretical relationship between bile flow, solvent drag, and inert solute diffusion predicted at extremely low bile flows, but demonstrated that the skate biliary tree is more permeable to inulin than that of the rat. Electron microscopic studies demonstrated that ionic lanthanum could traverse the tight junctions. However, freeze-fracture studies of junction structure did not differ qualitatively from similar studies in the rat. Partial dependence of bile flow on perfusion pressure, high bile-to-plasma inulin ratios, and permeability of the canalicular tight junctions to ionic lanthanum all suggest that the paracellular pathway may be an important component of bile formation in the skate.

scholarly journals Structural and Functional Regulation of Tight Junctions by RhoA and Rac1 Small GTPases

1998 ◽  
Vol 142 (1) ◽  
pp. 101-115 ◽  
Author(s):  
Tzuu-Shuh Jou ◽  
Eveline E. Schneeberger ◽  
W. James Nelson
Keyword(s):  
Tight Junctions ◽  
Spatial Organization ◽  
Mdck Cells ◽  
Protein Localization ◽  
Freeze Fracture ◽  
Tracer Diffusion ◽  
Small Gtpases ◽  
Dominant Negative ◽  
Solute Diffusion ◽  
Fence Function

Tight junctions (TJ) govern ion and solute diffusion through the paracellular space (gate function), and restrict mixing of membrane proteins and lipids between membrane domains (fence function) of polarized epithelial cells. We examined roles of the RhoA and Rac1 GTPases in regulating TJ structure and function in MDCK cells using the tetracycline repressible transactivator to regulate RhoAV14, RhoAN19, Rac1V12, and Rac1N17 expression. Both constitutively active and dominant negative RhoA or Rac1 perturbed TJ gate function (transepithelial electrical resistance, tracer diffusion) in a dose-dependent and reversible manner. Freeze-fracture EM and immunofluoresence microscopy revealed abnormal TJ strand morphology and protein (occludin, ZO-1) localization in RhoAV14 and Rac1V12 cells. However, TJ strand morphology and protein localization appeared normal in RhoAN19 and Rac1N17 cells. All mutant GTPases disrupted the fence function of the TJ (interdomain diffusion of a fluorescent lipid), but targeting and organization of a membrane protein in the apical membrane were unaffected. Expression levels and protein complexes of occludin and ZO-1 appeared normal in all mutant cells, although ZO-1 was more readily solubilized from RhoAV14-expressing cells with Triton X-100. These results show that RhoA and Rac1 regulate gate and fence functions of the TJ, and play a role in the spatial organization of TJ proteins at the apex of the lateral membrane.

Light and Electron Microscopic Studies Regarding Cell Contractility and Cell Coupling in Light Sensitive Smooth Muscle Cells from the Isolated Frog Iris Sphincter

1987 ◽  
Vol 42 (7-8) ◽  
pp. 977-985 ◽  
Author(s):  
Klaus V. Wolf
Keyword(s):  
Gap Junctions ◽  
Freeze Fracture ◽  
Muscle Cells ◽  
Sphincter Muscle ◽  
Cell Coupling ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Cell Contractility ◽  
Microscopic Studies ◽  
Iris Sphincter

(1) In light microscopical studies of living isolated frog irises, it was found that the maximal areas of experimentally light induced contractions in the m. sphincter pupillae were located beneath small illuminated regions. There were no visible contractions of muscle cells outside the illuminated areas. It was shown that exposure to light could directly cause contractions of isolated single sphincter muscle cells. (2) Junctional structures of the iris sphincter cells were studied by means of thin sections and freeze fracture electron microscopy. Intermediate junctions, a few focal tight junctions and occasional small gap junctions were identified. Pit containing intramembranous particles which resemble gap junction connexons were found in large numbers, dispersed over the plasmalemmas of sphincter muscle cells. From these physiological and morphological observations, it is concluded that sphincter muscle cells of the frog iris may be coupled via gap junctions, but that the cell coupling is not sufficiently extensive to form the basis for a functional syncytium.

Bile secretory function of intrahepatic biliary epithelium in the rat

1989 ◽  
Vol 257 (1) ◽  
pp. G124-G133 ◽  
Author(s):  
G. Alpini ◽  
R. Lenzi ◽  
W. R. Zhai ◽  
P. A. Slott ◽  
M. H. Liu ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Biliary Obstruction ◽  
Bile Flow ◽  
Secretory Activity ◽  
Biliary Tree ◽  
Cell Hyperplasia ◽  
Bile Formation ◽  
Biliary Epithelium ◽  
Biliary Epithelial Cells ◽  
Ductular Cell

To shed light on ductular fluid secretion, hepatic histology and ultrastructure, cell proliferation and phenotypes, and several aspects of biliary physiology were studied in rats with ductular cell hyperplasia induced by either biliary obstruction (0-14 days) or 1-naphthylisothiocyanate (ANIT) feeding (0-28 days). In both groups of experimental animals, bile duct hyperplasia and spontaneous bile flow and secretin-induced choleresis increased with time of treatment in a linear fashion. Measurements of [14C]mannitol biliary entry and of biliary tree volume showed that the increase in both spontaneous and secretin-stimulated bile flow originated at the proliferated biliary structures. Ultrastructural examination, [3H]thymidine incorporation, and histochemical and immunohistochemical staining for various markers demonstrated that in both hyperplastic reactions the proliferated cells were the progeny of preexisting biliary epithelial cells and retained their characteristics. These results indicate that the increased bile secretory activity associated with either biliary obstruction or ANIT intoxication reflects a quantitative change due to the proliferation of biliary epithelial cells. Thus both models of bile ductular cell hyperplasia lend themselves to assessment of the transport function of intrahepatic biliary epithelium and its contribution to normal bile formation. In the present studies, we have estimated that net ductular secretion in the normal rat accounts for 10-13% of spontaneously secreted hepatic bile.

Bile secretion of sulfated glycolithocholic acid is required for its cholestatic action in rats

1992 ◽  
Vol 262 (2) ◽  
pp. G267-G273
Author(s):  
F. Kuipers ◽  
M. J. Hardonk ◽  
R. J. Vonk ◽  
R. van der Meer
Keyword(s):  
Intravenous Injection ◽  
Biliary Drainage ◽  
Bile Flow ◽  
Wistar Rats ◽  
Parent Compound ◽  
Biliary Tree ◽  
Bile Secretion ◽  
Biliary Secretion ◽  
Contrast Administration ◽  
Bile Formation

To test our hypothesis that the cholestatic action of sulfated glycolithocholic acid (SGLC) in the rat is related to its interaction with calcium in the biliary tree [R. van der Meer, R. J. Vonk, and F. Kuipers. Am. J. Physiol. 254 (Gastrointest. Liver Physiol. 17): G644-G649, 1988], we have now compared its effects on bile formation in control Wistar rats and mutant Groningen Yellow (GY) Wistar rats. Intravenous injection of 0.6 mumol/100 g body wt of [14C]SGLC in unanesthetized rats with permanent biliary drainage did not induce cholestasis in either of the strains; however, its biliary secretion was strongly impaired in GY rats (12% dose at 1 h after injection vs. 95% dose in controls). Injection of 6.0 and 12.0 mumol/100 g body wt of [14C]SGLC caused an almost complete cessation of bile flow in control rats within 3 and 1 h, respectively. In contrast, administration of the same doses did not cause cholestasis in GY rats. Cholestasis in control rats was preceded by coprecipitation of [14C]SGLC and calcium in bile and incomplete biliary recovery of radioactivity. The hepatic content 15 min after injection of [14C]SGLC (6.0 mumol/100 g body wt) was similar in control and GY rats, 51 and 49% of the dose, respectively. Administration of glycolithocholic acid, the unsulfated parent compound of SGLC (6.0 mumol/100 g body wt), induced a rapid but reversible cessation of bile flow in both controls and GY rats; in this case no precipitation was observed in bile. This study shows that rapid bile secretion of SGLC is required for the induction of cholestasis.(ABSTRACT TRUNCATED AT 250 WORDS)

Experimental Disruption of the Blood-Brain Barrier System in an Insect (Periplaneta Americana)

1973 ◽  
Vol 59 (3) ◽  
pp. 711-724
Author(s):  
J. E. TREHERNE
Keyword(s):  
Blood Brain Barrier ◽  
Periplaneta Americana ◽  
Brain Barrier ◽  
Ultrastructural Changes ◽  
Hypertonic Solution ◽  
Electron Microscopic ◽  
Blood Brain ◽  
Ionic Lanthanum ◽  
Electrophysiological Studies ◽  
Microscopic Studies

1. The effects of hypertonic urea and hypertonic glucose solutions upon the ‘blood-brain barrier’ in the isolated abdominal nerve cord of the cockroach have been studied. 2. Electrophysiological studies showed that a hypertonic solution of urea, but not of glucose, was effective in reducing the barrier to the entry of potassium and the loss of sodium. 3. Electron-microscopic studies revealed no significant ultrastructural changes, and no disruption of the perineurial barrier to the entry of ionic lanthanum, following comparable exposure to the hypertonic solutions. 4. It is suggested that this alteration of the blood-brain barrier may result from a selective change in permeability of the perineurial membranes or tight junctions.

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