Demonstration of protein asymmetries in the plasma membrane of cultured renal (MDCK) epithelial cells by lactoperoxidase‐mediated iodination

Richardson, J.C.; Simmons, Nicholas L.

doi:10.1016/0014-5793(79)80611-0

Cited by 74 publications

(42 citation statements)

References 15 publications

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“…Madin-Darby canine kidney (MDCK) cells are polarized epithelial cells (Cereijido et al, 1978). Intercellular tight junctions separate the plasma membrane into functionally distinct apical and basolateral domains, which differentially express a variety of proteins (Richardson & Simmons, 1979). When grown in permeable chambers, the plasma membrane is differentiated into an apical surface with microvilli facing the growth medium and a basolateral surface facing the neighbouring cells and the substratum.…”

Section: Introductionmentioning

confidence: 99%

Infection of a polarized epithelial cell line with wild-type reovirus leads to virus persistence and altered cellular function

Montgomery

Yang

Verdin

et al. 1991

Journal of General Virology

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Section: Introductionmentioning

confidence: 99%

Infection of a polarized epithelial cell line with wild-type reovirus leads to virus persistence and altered cellular function

Montgomery

Yang

Verdin

et al. 1991

Journal of General Virology

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“…These cells display both structural and functional polarity when grown in culture (24,32). The microvillar THE JOURNAL OF CELL BIOLOGY .…”

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Transepithelial transport of a viral membrane glycoprotein implanted into the apical plasma membrane of Madin-Darby canine kidney cells. I. Morphological evidence.

Matlin¹,

Bainton²,

Pesonen³

et al. 1983

The Journal of Cell Biology

104

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The G protein of vesicular stomatitis virus was implanted in the apical plasma membrane of Madin-Darby canine kidney cells by low pH-dependent fusion of the viral envelope with the cellular membrane. The amount of fusion as determined by removal of unfused virions, either by tryptic digestion or by EDTA treatment at 0°C, was 22-24% of the cell-bound virus radioactivity. Upon incubation of cells after implantation, the amount of G protein as detected by immunofluorescence diminished on the apical membrane and appeared within 30 min on the basolateral membrane. At the same time some G protein fluorescence was also seen in intracellular vacuoles. The observations by immunofluorescence were confirmed and extended by electron microscopy. Using immunoperoxidase localization, G protein was seen to move into irregularly shaped vacuoles (endosomes) and multivesicular bodies and to appear on the basolateral plasma membrane. These results suggest that the apical and basolateral domains of Madin-Darby canine kidney cells are connected by an intracellular route.In most cells the plasma membrane is continuously endocytosed (36) and the loss of surface membrane must be balanced by retrieval of membrane from inside the cell. Membrane recycling poses a special problem in epithelial cells because the plasma membrane is polarized; it is divided into apical and basolateral domains with distinct protein and lipid compositions (35). If the plasma membrane is continuously recycling, an important question is how the apical and basolateral domains preserve their unique compositions. If the apical and the basolateral recycling routes are separated, randomization of the cell surface would of course be prevented. If, however, they connect at some point in the cell, continuous sorting of apical and basolateral proteins would have to take place.In an attempt to map the membrane traffic routes to and from the cell surface in epithelial cells, we are using the MadinDarby canine kidney (MDCK) cell line as our experimental system (7, 16). These cells display both structural and functional polarity when grown in culture (24, 32). The microvillar

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“…Like natural epithelia, MDCK cells generate two domains in the plasma membrane, apical and basolateral, separated by tight junctions (1,2,8,12). Plasma membrane proteins are asymmetrically distributed between the apical and basolateral surfaces (10,16). This is dramatically demonstrated by the asymmetric budding of enveloped viruses from only one plasma membrane domain, which results from the accumulation of viral envelope glycoproteins in the appropriate plasma membrane region (19,20).…”

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Lectin-resistant mutants of polarized epithelial cells.

Meiss¹,

Green²,

Rodríguez-Boulan³

1982

Mol. Cell. Biol.

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Two lectin-resistant mutants derived from Madin Darby canine kidney cells, with constitutive alterations in the asparagine-linked carbohydrate moieties, retained the characteristic structural and functional epithelial polarity of the parental cells. A ricin-resistant cell line was unable to incorporate galactose-sialic acid into glycoproteins and, from the pattern of cross-resistance to other lectins, appears to be different from previously described lines resistant to this lectin; the mutation in a concanavalin A-resistant line results, probably, in the production of defective carbohydrate cores of glycoproteins. In spite of glycosylation defects which result in an increased electrophoretic mobility of many cellular glycoproteins, both mutants retained the typical asymmetric structure of the plasma membrane (microvilli on the apical surface, junctional elements on the basolateral surface), functional tight junctions, and unidirectional active transport of electrolytes and water. These results suggest that glycoproteins with terminal galactosesialic acid moieties are not critically involved in the development and maintenance of polarity in epithelial cells. The mutant cells, particularly the ricin-resistant line, exhibited, however, morphological and electrophysiological changes which suggest a quantitative effect of the mutations on intracellular traffic of membranes and tight junction formation. The cell lines described in this paper, the first lectinresistant mutants of epithelial lineage, should prove useful tools for studying the peculiarities of glycosylating pathways in polarized cells.

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Demonstration of protein asymmetries in the plasma membrane of cultured renal (MDCK) epithelial cells by lactoperoxidase‐mediated iodination

Cited by 74 publications

References 15 publications

Infection of a polarized epithelial cell line with wild-type reovirus leads to virus persistence and altered cellular function

Infection of a polarized epithelial cell line with wild-type reovirus leads to virus persistence and altered cellular function

Transepithelial transport of a viral membrane glycoprotein implanted into the apical plasma membrane of Madin-Darby canine kidney cells. I. Morphological evidence.

Lectin-resistant mutants of polarized epithelial cells.

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