Caco-2 cells were cultured on permeable supports. At confluence the minute-by-minute net water movement (Jw) was automatically recorded. Simultaneously, unidirectional [14C]mannitol, 22Na+, and/or 36Cl- fluxes and transepithelial resistances were measured. The water and mannitol permeabilities went progressively down between 9 and 16 days after seeding and then stabilized. In this last condition the hydrostatic permeability coefficient (Phydr) was 2.67 +/- 0.31 cm s-1 while the osmotic permeability coefficient (Posm) was 0.0017 +/- 0.0004 cm s-1. Phydr but not Posm was dependent on the temperature and on the presence of Na+ in the medium. A net secretory Jw was observed 16 days after seeding, in the absence of any osmotic, hydrostatic or chemical gradient. This secretory Jw was associated with net Cl- (1.43 +/- 0.43 muequiv h-1 cm-2) and Na+ (1.05 +/- 0.35 muequiv h-1 cm-2) secretions. Amiloride reduced, in open-circuit conditions, both Na+ and Cl- apical to basal fluxes, thus enhancing the net Na+ and Cl- exit. Acidification of the medium (pH 6.2) reversibly increased water and mannitol permeabilities in 10-day-old cultures. In 16-day-old cultures the same shift in medium pH did not change mannitol permeability, while stimulating water secretion. These results, obtained in the absence of supracellular structures (villae, crypts) and subepithelial components (muscular, vascular and conjunctive tissues) indicate that paracellular and transport-associated water pathways are sensitive to changes in the pH of the medium in Caco-2 cell layers.
The minute-by-minute net water movement (Jw) in the rat cecum was correlated with the transepithelial potential difference (PD), short-circuit current (Isc), and the unidirectional Na+, Cl-, and Rb+ fluxes, with the following results. 1) Jw was a linear function of the applied hydrostatic or osmotic transepithelial gradients (hydrostatic permeability coefficiency = 0.164 +/- 0.018 cm/s, n = 13; osmotic permeability coefficient = 0.0014 +/- 0.0002 cm/s, n = 6). 2) A fraction of this absorptive Jw (0.17 +/- 0.03 microliter.min-1.cm-2, n = 13) was independent of the presence of any osmotic, hydrostatic, or chemical gradient. 3) This fraction was Na+ dependent, associated with an amiloride-insensitive PD and net Na+ (2.37 +/- 0.68 mu eq.h-1.cm-2, n = 6) and Cl- influxes (3.45 +/- 1.46 mu eq.h-1.cm-2, n = 6), measured under short-circuit conditions. No net Rb+ movement was detected. 4) The absorptive Jw increased when HCO3- was replaced by tris(hydroxymethyl)aminomethane (Tris+) buffer or Cl- by SO4(2-). A good agreement between the observed and the expected Jw (assuming isosmotic reabsorption) was observed in the absence of HCO3-. 5) The presence of an osmotic but not a hydrostatic transepithelial gradient generated a transepithelial PD. These results show that water movement across the rat cecum in vitro is the result of a combination of hydrostatic-, osmotic-, and transport-associated transfers. Concerning this last driving force, the observed results indicate that the transport-related Jw results from the addition of an absorptive Jw, coupled to a nonelectrogenic NaCl entry, plus a secretory Jw probably coupled to HCO3- secretion.
The water permeability properties of a series of epithelial barriers (the toad urinary bladder [TUB], the rat caecum [RC], the distal human colon [DHC], and the human amnion [HA] were studied in different experimental conditions. Three parameters were simultaneously determined: the water permeability coefficient in the presence of a transepithelial hydrostatic gradient (Phydr); the water permeability coefficient in the presence of an osmotic gradient (Posm); and the transepithelial potential difference (dV). All experiments were performed with the same experimental device, allowing comparison of the permeability properties of the barriers tested. The results obtained were: (1) TUB (N = 8): Phydr = 0.079 +/- 0.008 cm/s; Posm = 0.0004 +/- 0.0002 cm/s; dV = 31 +/- 5 mV; (2) TUB after ADH (N = 8): Phydr = 0.093 +/- 0.012 cm/s; Posm = 0.0065 +/- 0.0011 cm/s; dV = 52 +/- 8; (3) RC (N = 10): Phydr = 0.18 +/- 0.02 cm/s; Posm = 0.0019 +/- 0.0004 cm/s; dV = 3.9 +/- 0.1 mV; (4) RC adapted to a high K diet (N = 10): Phydr = 0.21 +/- 0.02 cm/s; Posm = 0.0018 +/- 0.0006 cm/s; dV = 4.5 +/- 0.5 mV; (5) DHC (N = 6): Phydr = 0.22 +/- 0.03 cm/s; Posm = 0.002 +/- 0.05 cm/s; dV = 15 +/- 3 mV; (6) HA (N = 10): Phydr = 0.32 +/- 0.05 cm/s; Posm = 0.0154 +/- 0.0015; dV = 0. The results show a good correlation between Phydr and dV, but not between dV and Posm or between Posm and Phydr.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.