Isolation of the basal and lateral plasma membranes of rat kidney tubule cells

Ebel, H.; Aulbert, E.; Merker, H. J.

doi:10.1016/0005-2736(76)90279-0

Cited by 37 publications

(7 citation statements)

References 47 publications

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“…Scalera, Yu-Kuo Huang, B. Hildmnn, and H . Murer fractionation by free-flow electrophoresis (5) or with density gradient centrifugations (3,8,11). They are time-consuming and involve equipment not easily available.…”

Section: Introductionmentioning

confidence: 99%

A Simple Isolation Method for Basal-Lateral Plasma Membranes from Rat Kidney Cortex

Scalera

Huang

Hildmann

et al. 1981

Membrane Biochemistry

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Basal-lateral membranes were separated in a self-orienting Percoll (modified colloidal silica) gradient from a heavy microsomal membrane fraction by centrifugation at 48,000g for 0.5 h. The (Na+--K+)-ATPase activity as a marker enzyme for the basal-lateral plasma membrane was 20-fold enriched by this procedure. The adenylate-cyclase activity measured in the basal-lateral membrane fraction was stimulated 6-fold by parathyrin and only up to 1.5-fold by arginine-vasopressin, calcitonin, or isoproterenol. The yield of basal-lateral plasma membranes was 5 to 10 percent of the amount initially present in the homogenate. The method is also applicable to the pig kidney.

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Section: Introductionmentioning

confidence: 99%

A Simple Isolation Method for Basal-Lateral Plasma Membranes from Rat Kidney Cortex

Scalera

Huang

Hildmann

et al. 1981

Membrane Biochemistry

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“…Structural polarity has also been obseved by freeze-fracture electron microscopy with the intramembrane particle density being higher on the basal-lateral than the apical plasma membrane (3)(4)(5). Biochemical studies of purified apical and basal-lateral mem-THE JOURNAL OF CELL BIOLOGY -VOLUME 98 MAY 1984 1777-1787 0 The Rockefeller University Press -0021-9525/84/05/1777/11 $1 .00 brane preparations from renal proximal tubule and small intestine have shown that these plasma membrane regions differ in composition with each membrane domain having their own characteristic set of enzymes, polypeptides, and lipids (6)(7)(8)(9)(10)(11) . The asymmetric distribution of cell surface components has been confirmed by autoradiography and immunocytochemistry (12)(13)(14)(15)(16) .…”

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confidence: 99%

Studies on the development and maintenance of epithelial cell surface polarity with monoclonal antibodies.

Herzlinger¹,

Ojakian²

1984

The Journal of Cell Biology

123

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We examined epithelial cell surface polarity in subconfluent and confluent MadinDarby canine kidney (MDCK) cells with monoclonal antibodies directed against plasma membrane glycoproteins of 35,000, 50,000, and 60,000 mol wt. The cell surface distribution of these glycoproteins was studied by immunofluorescence and immunoelectron microscopy. At the ultrastructural level, the electron-dense reaction product localizing all three glycoproteins was determined to be uniformly distributed over the apical and basal cell surfaces of subconfluent MDCK cells as well as on the lateral surfaces between contacted cells; however, after formation of a confluent monolayer, these glycoproteins could only be localized on the basal-lateral plasma membrane. The development of cell surface polarity was followed by assessing glycoprotein distribution with immunofluorescence microscopy at selected time intervals during growth of MDCK cells to form a confluent monolayer . These results were correlated with transepithelial electrical resistance measurements of tight junction permeability and it was determined by immunofluorescence that polarized distributions of cell surface glycoproteins were established just after electrical resistance could be detected, but before the development of maximal resistance . Our observations provide evidence that intact tight junctions are required for the establishment of the apical and basal-lateral plasma membrane domains and that development of epithelial cell surface polarity is a continuous process .The specialized functions of epithelia such as absorption, secretion, and vectorial ion transport are dependent on the existence of two distinct cell surfaces; the apical (or mucosal) domain which is separated from the basal-lateral (or serosal) surface by a circumferential occluding belt of tight junctions (1). These two membrane domains are in contact with different environments and each exhibits unique structural and biochemical components that are responsible for the specialized physiological properties of epithelia .Ultrastructural examination of a variety of epithelial tissues has demonstrated structural polarity with plasma membrane specializations such as microvilli and cilia being confined to the apical cell surface while intercellular junctions are localized to the basal-lateral membrane (1, 2). Structural polarity has also been obseved by freeze-fracture electron microscopy with the intramembrane particle density being higher on the basal-lateral than the apical plasma membrane (3-5). Biochemical studies of purified apical and basal-lateral mem-

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“…Seven marker enzymes (see Table 4) were measured by commonly used methods; NADPH-cytochrome c reductase (Sottocasa et al, 1967); glucose 6-phosphatase [Swanson, 1950, as modified by Hubscher & West (1965) as well as by Leighton et al (1968)]; succinate dehydrogenase (Brdiczka et al, 1968); lactate dehydrogenase (Bergmeyer & Bernt, 1974); acid phosphatase [Leighton et al (1968), but with p-nitrophenol as substrate as described by Walter & Schutt (1974)]; 5'-nucleotidase (Ebel et al, 1976); uricase [Bergmeyer (1974) but with 0.1 % Triton X-100 in the incubation mixtures as described by Leighton et al (1968)]. Protein was determined by the biuret (Bode et al, 1968) and the Lowry (Lowry et al, 1951) procedures, the latter with the addition of a constant quantity of dithioerythritol in standards and blanks to account for the contents of thiol groups.…”

Section: Methodsmentioning

confidence: 99%

Re-examination of the subcellular localization of thyroxine 5′-deiodination in rat liver

Brinke¹,

Hesch²,

Köhrle³

1979

Biochemical Journal

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We describe the existence of at least two thyroxine 5′-deiodinases in rat liver. They co-fractionate with NADPH-cytochrome c reductase, the marker enzyme for membranes of the endoplasmic reticulum. Subcellular-localization studies of the most active microsomal thyroxine 5′-deiodinase were performed under substrate saturation and at optimal pH 6.8. This enzyme was a Km(app.) of about 3 microM-thyroxine and a Vmax. of about 8 ng of tri-iodothyronine/min per mg of protein. Our study confirms in part the earlier reports of microsomal localization of thyroxine 5′-deiodination. However, this process is not mediated by only a single enzyme.

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Isolation of the basal and lateral plasma membranes of rat kidney tubule cells

Cited by 37 publications

References 47 publications

A Simple Isolation Method for Basal-Lateral Plasma Membranes from Rat Kidney Cortex

A Simple Isolation Method for Basal-Lateral Plasma Membranes from Rat Kidney Cortex

Studies on the development and maintenance of epithelial cell surface polarity with monoclonal antibodies.

Re-examination of the subcellular localization of thyroxine 5′-deiodination in rat liver

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