Acidic pH of the lateral intercellular spaces of MDCK cells cultured on permeable supports

1994 ◽  
Vol 140 (2) ◽  
Author(s):  
J.-Y. Chatton ◽  
K.R. Spring
Keyword(s):  
Mdck Cells ◽  
Acidic Ph ◽  
Intercellular Spaces ◽  
Lateral Intercellular Spaces ◽  
Permeable Supports ◽  
Cells Cultured

pH, morphology, and diffusion in lateral intercellular spaces of epithelial cell monolayers

1994 ◽  
Vol 266 (1) ◽  
pp. C73-C80 ◽  
Author(s):  
P. J. Harris ◽  
J. Y. Chatton ◽  
P. H. Tran ◽  
P. M. Bungay ◽  
K. R. Spring
Keyword(s):  
Diffusion Coefficient ◽  
Mdck Cells ◽  
Transepithelial Transport ◽  
Osmotic Gradient ◽  
Intercellular Spaces ◽  
Cell Monolayers ◽  
Lateral Intercellular Spaces ◽  
Epithelial Cell Monolayers ◽  
And Diffusion ◽  
Measuring Ph

The lateral intercellular spaces (LIS) of reabsorptive epithelia are the site of the proposed local osmotic gradient responsible for transepithelial transport. We developed techniques for loading the LIS of living cultured renal cells (MDCK and LLC-PK1) with the fluorescent dye 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF), visualizing LIS geometry, measuring pH, and determining the BCECF diffusion coefficient within the LIS. The LIS pH was remarkably constant and differed substantially from that of the superfusate in both the presence and absence of HCO3 or CO2. The LIS of MDCK cells had a pH of 7.66 +/- 0.04 in bicarbonate-free solutions of pH 7.0, 7.4, or 7.8. In bicarbonate-containing solutions, MDCK LIS pH was acidic to the superfusate by 0.3-0.4 units. In the absence of bicarbonate, the LIS of LLC-PK1 cells was markedly acidic (6.83 +/- 0.05), becoming alkaline by approximately 0.25 units in the presence of bicarbonate. Gradients in pH or dye concentration were not detected within the LIS. The diffusion coefficient of BCECF within the LIS was approximately equal to that seen in free solution.

scholarly journals Pseudo-streaming potentials in Necturus gallbladder epithelium. II. The mechanism is a junctional diffusion potential.

1992 ◽  
Vol 99 (3) ◽  
pp. 317-338 ◽  
Author(s):  
L Reuss ◽  
B Simon ◽  
C U Cotton
Keyword(s):  
Time Course ◽  
Bathing Solution ◽  
Intercellular Spaces ◽  
Similar Time ◽  
Streaming Potentials ◽  
Osmotic Water ◽  
Lateral Intercellular Spaces ◽  
Transepithelial Voltage ◽  
Time Courses ◽  
Good Agreement

The mechanisms of apparent streaming potentials elicited across Necturus gallbladder epithelium by addition or removal of sucrose from the apical bathing solution were studied by assessing the time courses of: (a) the change in transepithelial voltage (Vms). (b) the change in osmolality at the cell surface (estimated with a tetrabutylammonium [TBA+]-selective microelectrode, using TBA+ as a tracer for sucrose), and (c) the change in cell impermeant solute concentration ([TMA+]i, measured with an intracellular double-barrel TMA(+)-selective microelectrode after loading the cells with TMA+ by transient permeabilization with nystatin). For both sucrose addition and removal, the time courses of Vms were the same as the time courses of the voltage signals produced by [TMA+]i, while the time courses of the voltage signals produced by [TBA+]o were much faster. These results suggest that the apparent streaming potentials are caused by changes of [NaCl] in the lateral intercellular spaces, whose time course reflects the changes in cell water volume (and osmolality) elicited by the alterations in apical solution osmolality. Changes in cell osmolality are slow relative to those of the apical solution osmolality, whereas lateral space osmolality follows cell osmolality rapidly, due to the large surface area of lateral membranes and the small volume of the spaces. Analysis of a simple mathematical model of the epithelium yields an apical membrane Lp in good agreement with previous measurements and suggests that elevations of the apical solution osmolality elicit rapid reductions in junctional ionic selectivity, also in good agreement with experimental determinations. Elevations in apical solution [NaCl] cause biphasic transepithelial voltage changes: a rapid negative Vms change of similar time course to that of a Na+/TBA+ bi-ionic potential and a slow positive Vms change of similar time course to that of the sucrose-induced apparent streaming potential. We conclude that the Vms changes elicited by addition of impermeant solute to the apical bathing solution are pseudo-streaming potentials, i.e., junctional diffusion potentials caused by salt concentration changes in the lateral intercellular spaces secondary to osmotic water flow from the cells to the apical bathing solution and from the lateral intercellular spaces to the cells. Our results do not support the notion of junctional solute-solvent coupling during transepithelial osmotic water flow.

Hypertonicity stimulates transcription of gene for Na(+)-myo-inositol cotransporter in MDCK cells

1993 ◽  
Vol 264 (1) ◽  
pp. F20-F23 ◽  
Author(s):  
A. Yamauchi ◽  
S. Uchida ◽  
A. S. Preston ◽  
H. M. Kwon ◽  
J. S. Handler
Keyword(s):  
Mdck Cells ◽  
Renal Medulla ◽  
Mrna Abundance ◽  
Transport Rate ◽  
Madin Darby Canine Kidney ◽  
Hypertonic Medium ◽  
Compatible Osmolyte ◽  
Darby Canine Kidney ◽  
Cells Cultured ◽  
Canine Kidney

Myo-inositol is a major compatible osmolyte accumulated in the hypertonic renal medulla and in Madin-Darby canine kidney (MDCK) cells cultured in hypertonic media. Myo-inositol is taken up by MDCK cells on a Na(+)-coupled transporter whose activity increases sixfold 24 h after cells are switched to hypertonic medium. To investigate the mechanism of regulation of the cotransporter by hypertonicity, we used the cDNA encoding the canine Na(+)-myo-inositol cotransporter that we recently cloned to measure the abundance of the mRNA for the cotransporter and its rate of transcription after changes in osmolality. When MDCK cells were switched from isotonic to hypertonic medium, cotransporter mRNA abundance rose 10-fold in 16 h. Transcription of the cotransporter gene also rose and 16 h after the switch reached a peak approximately 15-fold that in isotonic cells. When cells were switched back to isotonic medium, mRNA abundance and transcription of the gene returned to isotonic levels in 8 h and transport rate reached isotonic levels in 48 h. Thus transcription appears to be the primary step in regulation of myo-inositol transport by hypertonicity.

scholarly journals Studies on the Electrical Potential Profile across Rabbit Ileum

1971 ◽  
Vol 57 (6) ◽  
pp. 639-663 ◽  
Author(s):  
Richard C. Rose ◽  
Stanley G. Schultz
Keyword(s):  
Amino Acids ◽  
Brush Border ◽  
Electrical Potential ◽  
Metabolic Inhibitors ◽  
Electrical Potential Difference ◽  
Intercellular Spaces ◽  
Cell Interior ◽  
Rabbit Ileum ◽  
Lateral Intercellular Spaces ◽  
Rapid Replacement

When isolated strips of mucosal rabbit ileum are bathed by physiological electrolyte solution the electrical potential difference (PD) across the brush border (ψmc) averages 36 mv, cell interior negative. Rapid replacement of Na in the mucosal solution with less permeant cations, Tris or choline, results in an immediate hyperpolarization of ψmc. Conversely, replacement of choline in the mucosal solution with Na results in an abrupt depolarization of ψmc. These findings indicate that Na contributes to the conductance across the brush border. The presence of actively transported sugars or amino acids in the mucosal solution brings about a marked depolarization of ψmc and a smaller increase in the transmural PD (Δψms). It appears that the Na influx that is coupled to the influxes of amino acids and sugars is electrogenic and responsible for the depolarization of ψmc. Under control conditions Δψms can be attributed to the depolarization of ψmc together with the presence of a low resistance transepithelial shunt, possibly the lateral intercellular spaces. However, quantitatively similar effects of amino acids on ψmc are also seen in tissues poisoned with metabolic inhibitors or ouabain. Under these conditions Δψmc is much smaller than under control conditions. Thus, the depolarization of ψmc might not account for the entire Δψms, observed in nonpoisoned tissue. An additional electromotive force which is directly coupled to metabolic processes might contribute to the normal Δψms.

Size Variations in the Lateral Intercellular Spaces of the Endolymphatic Sac Induced by Dietary Factors

1990 ◽  
Vol 100 (3) ◽  
pp. 217???222 ◽  
Author(s):  
Phillip A. Wackym ◽  
Rinaldo F. Canalis ◽  
Ulla Friberg ◽  
Helge Rask-Andersen ◽  
Fred H. Linthicum
Keyword(s):  
Dietary Factors ◽  
Endolymphatic Sac ◽  
Intercellular Spaces ◽  
Lateral Intercellular Spaces
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