Secretion of endogenous and exogenous proteins from polarized MDCK cell monolayers.
Confluent monolayers of MDCK (Madin-Darby canine kidney) cells provide a widely used system to study the biogenesis of epithelial cell polarity. We now report that these cells are also capable of the vectorial constitutive secretion of a major endogenous product, a glycoprotein of 81 kDa, which is released into the medium from the apical surface within 30 min of its synthesis. This release represents a bona fide exocytotic secretory process and is not the result of proteolytic cleavage of a plasma membrane-associated precursor since, in cells treated with chloroquine, a protein indistinguishable from the mature secretory product accumulated intracellularly. In contrast to the vectorial secretion of the endogenous product, a variety of exogenous exocrine and endocrine proteins synthesized in MDCK cells transfected with the corresponding genes were secreted from both the apical and basolateral surfaces. These included proteins such as rat growth hormone, chicken oviduct lysozyme, bovine gastric prochymosin, and rat salivary gland alpha 2u-globulin, which in their cells of origin are secreted via a regulated pathway, as well as the liver form of the alpha 2u-globulin and the immunoglobulin kappa chain, which are normally released constitutively. These results demonstrate the existence of secretory pathways that lead to both surfaces of MDCK cells and are accessible to the foreign secretory products. They are consistent with the operation of a sorting mechanism in which the polarized secretion of the endogenous product is effected through the recognition of signals that prevent its random distribution within the fluid phase in the cellular endomembrane system.
Gottlieb
T A
TA
Beaudry
G
G
Rizzolo
L
L
Colman
A
A
Rindler
M
M
Adesnik
M
M
Sabatini
D D
DD
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002087:Butyrates / Q000494:pharmacology; D002451:Cell Compartmentation / Q000187:drug effects; D002460:Cell Line; D002738:Chloroquine / Q000494:pharmacology; D003470:Culture Media; D004285:Dogs; D004848:Epithelium / Q000378:metabolism* / Q000648:ultrastructure; D006023:Glycoproteins / Q000378:metabolism*; D013006:Growth Hormone / Q000378:metabolism; D007145:Immunoglobulin kappa-Chains / Q000378:metabolism; D007668:Kidney; D007700:Kinetics; D008970:Molecular Weight; D009113:Muramidase / Q000378:metabolism; D012636:Secretory Rate / Q000187:drug effects
D002087:Butyrates; D003470:Culture Media; D006023:Glycoproteins; D007145:Immunoglobulin kappa-Chains; D002738:Chloroquine; D013006:Growth Hormone; D009113:Muramidase