The polarized expression of Na+,K+-ATPase in epithelia depends on the association between beta-subunits located in neighboring cells.
The polarized distribution of Na+,K+-ATPase plays a paramount physiological role, because either directly or through coupling with co- and countertransporters, it is responsible for the net movement of, for example, glucose, amino acids, Ca2+, K+, Cl-, and CO3H- across the whole epithelium. We report here that the beta-subunit is a key factor in the polarized distribution of this enzyme. 1) Madin-Darby canine kidney (MDCK) cells (epithelial from dog kidney) express the Na+,K+-ATPase over the lateral side, but not on the basal and apical domains, as if the contact with a neighboring cell were crucial for the specific membrane location of this enzyme. 2) MDCK cells cocultured with other epithelial types (derived from human, cat, dog, pig, monkey, rabbit, mouse, hamster, and rat) express the enzyme in all (100%) homotypic MDCK/MDCK borders but rarely in heterotypic ones. 3) Although MDCK cells never express Na+,K+-ATPase at contacts with Chinese hamster ovary (CHO) cells, they do when CHO cells are transfected with beta1-subunit from the dog kidney (CHO-beta). 4) This may be attributed to the adhesive property of the beta1-subunit, because an aggregation assay using CHO (mock-transfected) and CHO-beta cells shows that the expression of dog beta1-subunit in the plasma membrane does increase adhesiveness. 5) This adhesiveness does not involve adherens or tight junctions. 6) Transfection of beta1-subunit forces CHO-beta cells to coexpress endogenous alpha-subunit. Together, our results indicate that MDCK cells express Na+,K+-ATPase at a given border provided the contacting cell expresses the dog beta1-subunit. The cell-cell interaction thus established would suffice to account for the polarized expression and positioning of Na+,K+-ATPase in epithelial cells.
Shoshani
Liora
L
Center for Research and Advanced Studies, Department of Physiology, Biophysics, and Neurosciences, México City 07300, México. shoshani@fisio.cinvestav.mx
Contreras
Rubén G
RG
Roldán
María L
ML
Moreno
Jacqueline
J
Lázaro
Amparo
A
Balda
María S
MS
Matter
Karl
K
Cereijido
Marcelino
M
D022005:Adherens Junctions; D000818:Animals; D015153:Blotting, Western; D016466:CHO Cells; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D018920:Coculture Techniques; D006224:Cricetinae; D004285:Dogs; D017097:Electric Impedance; D004848:Epithelium / Q000201:enzymology*; D005347:Fibroblasts / Q000378:metabolism; D015151:Immunoblotting; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012964:Sodium / Q000737:chemistry; D000254:Sodium-Potassium-Exchanging ATPase / Q000096:biosynthesis*; D019108:Tight Junctions; D014162:Transfection
D012964:Sodium; D000254:Sodium-Potassium-Exchanging ATPase