Effects of ouabain on the function and structure of a cell line (MDCK) derived from canine kidney

Abaza, Nabil A.; Leighton, Joseph; Schultz, Stanley G.

doi:10.1007/bf02615230

Cited by 75 publications

(29 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Yet, data in Table 3 show that the distribution of pumps is asymmetric enough to account for the net inward Na flux. This result is in keeping with the following facts: (i) MDCK monolayers cultured on nonpermeable supports produce blisters attributed to the accumulation of water (Leighton et al, 1969;Rabito etal., 1978); (ii) transport of water between isotonic media is not a primary phenomenon, but arises from coupling to solute transport; and (iii) blister formation in MDCK monolayers is inhibited by ouabain (Abaza, Leighton & Schultz, 1974). Table 5 compares the values obtained in MDCK monolayers with those of natural epithelia.…”

Section: Discussionmentioning

confidence: 93%

Structural and functional membrane polarity in cultured monolayers of MDCK cells

et al. 1980

View full text Add to dashboard Cite

MDCK cells form monolayers which have many of the properties usually found in transporting epithelia. The present article is devoted to the study of the structural and functional polarization of MDCK cells, which is one of the central features of transporting epithelia. The results show: (i) that MDCK monolayers transport 2.6 mumol hr-1 cm-2 of sodium in the apical to basolateral direction; (ii) the passive flux of this ion is relatively large (20.3 mole hr-1 cm-2), which is a characteristic of leaky epithelia; (iii) a large fraction of the penetration of sodium into the cells proceeds through an amiloride-sensitive channel, and the exit is operated mainly by a ouabain-sensitive pump; (iv) the net transport of sodium from the apical to the basolateral side agrees with the asymmetric labeling of the pumps with 3H-ouabain; (v) this asymmetric labeling agrees, in turn, with a higher concentration of intramembrane particles (IMPs) in freeze-fracture replicas of the basolateral side of the plasma membrane; (vi) the structural polarization of confluent MDCK cells is also revealed by the location of microvilli, occluding junctions, and pinocytotic vesicles; and (vii) the presence of a continuous ring formed by actin microfilaments visualized by immunofluorescence under the lateral aspect of the plasma membrane that may be related to the distribution of the occluding junctions, which act as barriers separating apical from basolateral membrane components.

show abstract

Section: Discussionmentioning

confidence: 93%

Structural and functional membrane polarity in cultured monolayers of MDCK cells

et al. 1980

View full text Add to dashboard Cite

show abstract

“…Morphological polarity, the presence of occluding junctions [14], and the reversible ouabain inhibition of hemicyst formation [1] constitute acceptable evidence that the hemicyst is an epithelium engaged in active transport. In addition, the small spontaneous A O and the practical absence of solute gradients across the hemicyst wall (see Table 2) means that it is an epithelium with a leaky junction.…”

Section: Discussionmentioning

confidence: 99%

“…They form and expand continuously, apparently under pressure of accumulating fluid. Finally, the hemicyst bursts and promptly collapses to reform again [1]. A small lateral diffusion is difficult to discard, but at least it is so small that it allows the hemicysts to form up to the point at which they burst.…”

Section: Discussionmentioning

confidence: 99%

“…Mg ++ and Mn +-had the same effect, while dbc-AMP, papaverine, and Ca ++ enhanced hemicyst formation [24]. Ouabainsensitive ATPase activity has been demonstrated by biochemical measurement [1]. All of these findings suggest that hemicyst formation in MDCK…”

mentioning

confidence: 88%

“…Since then hemicysts have been reported in other normal and neoplastic cell cultures [8,13,17,[20][21][22][23]. In each instance, the cells cultured were from transporting epithelia.Time lapse cinematographic study has demonstrated that the hemicysts in MDCK monolayers were dynamic structures [11], and that ouabain, an inhibitor of active solute transport, abolished hemicyst formation [1]. Mg ++ and Mn +-had the same effect, while dbc-AMP, papaverine, and Ca ++ enhanced hemicyst formation [24].…”

mentioning

confidence: 99%

See 2 more Smart Citations

Distribution and characteristics of the occluding junctions in a monolayer of a cell line (MDCK) derived from canine kidney

Rabito¹,

Tchao²,

Valentich³

et al. 1978

J. Membrain Biol.

Self Cite

View full text Add to dashboard Cite

On solid substrates MDCK, a cell line derived from normal dog kidney, forms a confluent monolayer that is studded with "blisters". Previous studies with this cell line suggest that these hemicysts develop as a result of active fluid accumulation between cell sheet and substratum. One factor that may determine when and how hemicysts appear only in localized sites is the interruption of occluding junctions in nonhemicyst areas. To study this possibility, we compared the permeability characteristics of the occluding junctions in hemicysts and in an uninterrupted monolayer of MDCK grown on a permeable support of collagen-coated nucleopore filter. The spontaneous electrical potential differences were small, without statistical differences between them. Relative ionic permeability coefficients were evaluated from the voltage deflections to imposed salt gradients or to a single ion substitution across both structures. The results showed that the relative permeability ratios for Na+, K+, choline+, and Cl- were the same in hemicysts and the uninterrupted monolayer. These and other results indicate that the junctional complex encircling the apical surface of a sheet of MDCK cells can provide an effective permeability barrier constituting a true occluding junction with the same properties in hemicyst and nonhemicyst areas.

show abstract

Characterization

Freshney¹

2005

Culture of Animal Cells

View full text Add to dashboard Cite

Effects of ouabain on the function and structure of a cell line (MDCK) derived from canine kidney

Cited by 75 publications

References 13 publications

Structural and functional membrane polarity in cultured monolayers of MDCK cells

Structural and functional membrane polarity in cultured monolayers of MDCK cells

Distribution and characteristics of the occluding junctions in a monolayer of a cell line (MDCK) derived from canine kidney

Characterization

Contact Info

Product

Resources

About