Methods for rapidly altering the permeability of mammalian cells

Heppel, Leon A.; Makan, Nizar R.

doi:10.1002/jss.400060313

Cited by 22 publications

(8 citation statements)

References 35 publications

(21 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Tweens (polyoxyethylene sorbitan esters) are nontoxic surfactants that are used extensively in the emulsification and solubilization of drugs and fat-soluble vitamins (for reviews, see 56,57). Tweens are believed to alter permeability reversibly and to allow the entry of various substances that are ordinarily unable to enter intact cells (58).…”

Section: Vitamin E Uptake By Intestinal Cellsmentioning

confidence: 99%

Transport of vitamin E by differentiated Caco-2 cells

Anwar

Kayden

Hussain

2006

Journal of Lipid Research

View full text Add to dashboard Cite

In hepatocytes, vitamin E is secreted via the efflux pathway and is believed to associate with apolipoprotein B (apoB)-lipoproteins extracellularly. The molecular mechanisms involved in the uptake, intracellular trafficking, and secretion of dietary vitamin E by the intestinal cells are unknown. We observed that low concentrations of Tween-40 were better for the solubilization and delivery of vitamin E to differentiated Caco-2 cells, whereas high concentrations of Tween-40 and sera inhibited this uptake. Vitamin E uptake was initially rapid and then reached saturation. Subcellular localization revealed that vitamin E primarily accumulated in microsomal membranes. Oleic acid (OA) treatment, which induces chylomicron assembly and secretion, decreased microsomal membrane-bound vitamin E in a time-dependent manner. To study secretion, differentiated Caco-2 cells were pulse-labeled with vitamin E and chased in the presence and absence of OA. In the absence of OA, vitamin E was associated with intestinal high density lipoprotein (I-HDL), whereas OAtreated cells secreted vitamin E with I-HDL and chylomicrons. No extracellular transfer of vitamin E between these lipoproteins was observed. Glyburide, an antagonist of ABCA1, partially inhibited its secretion with I-HDL, whereas plasma HDL increased vitamin E efflux. An antagonist of microsomal triglyceride transfer protein, brefeldin A, and monensin specifically inhibited vitamin E secretion with chylomicrons. These studies indicate that vitamin E taken up by Caco-2 cells is stored in the microsomal membranes and secreted with chylomicrons and I-HDL. Transport via I-HDL might contribute to vitamin E absorption in patients with abetalipoproteinemia receiving large oral doses of the vitamin. Vitamin E is a major lipid-soluble antioxidant and is an essential nutrient for normal growth and development. Its deficiency results in neurological dysfunction, muscular weakness, and reproductive failure (1-3). Although a diet rich in vegetable oils and whole grains is a sufficient source of vitamin E in normal people (4), deficiency can result because of lipid malabsorption syndromes such as abetalipoproteinemia, a disease in which apolipoprotein B (apoB)-containing lipoproteins (chylomicrons, very low density lipoproteins, and low density lipoproteins) are absent in the plasma (1, 2). High oral doses of vitamin E ameliorate the deficiency and normalize plasma and adipose tissue levels of the vitamin in these patients (5). The mechanisms involved in vitamin E absorption in the absence of apoB-lipoprotein assembly are not known.Current knowledge of intestinal vitamin E absorption is based on human and animal studies involving oral or intraduodenal administration of vitamin E followed by lymph and plasma analysis (1-3, 6-9). Vitamin E uptake by the intestinal cells is believed to be less efficient than that of other fat-soluble vitamins. In most studies, z20-50% of dietary vitamin E is believed to be absorbed (4, 10). In contrast, z80% of dietary vitamin A is absorbed in 24 h...

show abstract

Section: Vitamin E Uptake By Intestinal Cellsmentioning

confidence: 99%

Transport of vitamin E by differentiated Caco-2 cells

Anwar

Kayden

Hussain

2006

Journal of Lipid Research

View full text Add to dashboard Cite

show abstract

“…Because of their low lipid solubility the diffusion of these molecules through the lipid phase of the membrane is probably negligible (Plagemann & Wohlhueter, 1980). Cells can be rendered artificially permeable by various methods (Heppel & Makan, 1977) such as treatment with toluene (Moses & Richardson, 1970;Hilderman, Goldblatt & Deutscher, 1975), nonionic detergents (Kay, 1965;Malenkov et al, 1967;Carlson, Till & Ling, 1976), ionic detergents (Hodes, Palmer & Warren, 1960), osmotic shock (Kaltenbach, 1966;Castellot, Miller & Pardee, 1978), dextran sulfate (Kasahara, 1977) and exogenous ATP (Rozengurt & Heppel, 1975a). The latter is of special interest, since the increase in the passive permeability of the plasma membrane by exogenous ATP was found to be rapid, reversible, specific for ATP and selective for transformed cells (Rozengurt & Heppel, 1975a;Rozengurt, Heppel & Friedberg, 1977).…”

Section: Introductionmentioning

confidence: 99%

Permeability change in transformed mouse fibroblasts caused by ionophores, and its relationship to membrane permeabilization by exogenous ATP

Friedberg

Weisman

1985

J. Membrain Biol.

View full text Add to dashboard Cite

Electrogenic ionophores have been found to induce membrane permeabilization in Swiss mouse 3T3 cells that had undergone spontaneous transformation (3T6 cells). Cells attached to plastic dishes were loaded with [3H] uridine, and then the medium was replaced by buffered salt solution at pH 7.8. The enhancement of membrane permeability was assayed by following the efflux of uridine nucleotides, normally impermeant substances. Titration with electrogenic ionophores, such as carbonylcyanide m-chlorophenylhydrazone (CCCP), SF-6847 and gramicidin D, markedly increased the membrane permeability within a very narrow range of ionophore concentration. Non-electrogenic ionophores, such as monensin and nigericin, did not affect membrane permeability. Measurements of the distribution of the lipophilic cation tetraphenylphosphonium (TPP+) between the cells and their environment implied that the remarkable increase in permeability took place within a narrow range of membrane potential (delta psi). The data could be explained by a delta psi threshold value, under which aqueous channels are opened in the plasma membrane. The effects exerted by electrogenic ionophores on the plasma membrane were found to be similar to those induced by exogenous ATP. In both cases rapid efflux of K+, influx of Na+ and reduction of delta psi preceded membrane permeabilization to low molecular weight, charged molecules, such as nucleotides. It is suggested that dissipation of delta psi induces conformational alterations in membranal components, and/or topological changes, such as aggregation of protein molecules, to form membranal aqueous channels. Electrogenic ionophores permeabilize both normal (3T3) and transformed (3T6) mouse fibroblasts, whereas ATP effects are specific for transformed cells. Thus, it is postulated that ATP acts via specific sites on the surface of transformed cells.

show abstract

“…However, lower concentrations of these detergents can be used as valuable probes of cell membrane function in intact cells. At concentrations below those that solubilize membranes, non-ionic and zwitterionic detergents can be incorporated into the intact membrane, increase membrane fluidity, alter cell permeability, and affect in-situ membrane enzyme function (Helenius & Simons, 1975;Heppel & Makan, 1977 ;Gonenne& Ernst, 1978 ;Meeks& Chen, 1979;Juliano &Gagalong, 1979;Schmidt, Lichtenberg, Jackson & Litman, 1980). Accordingly, we have extended our earlier amino acid transport studies (DiZio & Tasca, 1977) by examining the effects of low concentrations of nonionic and zwitterionic detergents on preimplantation mouse embryos.…”

Section: Introductionmentioning

confidence: 99%

Modulation of amino acid transport in preimplantation mouse embryos by low concentrations of non-ionic and zwitterionic detergents

Keefer¹,

Tasca²

1984

Reproduction

View full text Add to dashboard Cite

In 4-cell embryos (but not in blastocysts), Triton X-100, a non-ionic detergent, stimulated leucine, phenylalanine, methionine and glutamic acid transport from 1.6 to 3.2-fold. All of these amino acids were transported exclusively by a sodium-independent mechanism. Triton X-100, however, did not stimulate the transport of other amino acids tested in 4-cell embryos. Furthermore, phenylalanine transport rates were stimulated about 2-fold at the 4-cell stage by all of the non-ionic and zwitterionic detergents tested at concentrations which were approximately one-tenth of the critical micellar concentration for each detergent. These concentrations did not block development, disrupt the cells, or make the cell membranes freely permeable. At the blastocyst stage, Z312, a zwitterionic detergent, inhibited the transport of phenylalanine and alanine and stimulated the transport of lysine, a pattern previously found to be linked to the sodium-dependent amino acid transport mechanism. We suggest that Z312 may be acting upon some component of sodium-dependent amino acid transport in blastocysts. The non-ionic and zwitterionic detergents seemed to have a common effect on amino acid transport in 4-cell embryos but elicited varied transport responses from blastocysts. These differential responses to detergents by blastocysts may reflect intrinsic changes in membrane composition and/or organization which occur during the normal course of preimplantation development.

show abstract

Methods for rapidly altering the permeability of mammalian cells

Cited by 22 publications

References 35 publications

Transport of vitamin E by differentiated Caco-2 cells

Transport of vitamin E by differentiated Caco-2 cells

Permeability change in transformed mouse fibroblasts caused by ionophores, and its relationship to membrane permeabilization by exogenous ATP

Modulation of amino acid transport in preimplantation mouse embryos by low concentrations of non-ionic and zwitterionic detergents

Contact Info

Product

Resources

About