[43] Permeabilizing mammalian cells to macromolecules

Weisman, Gary A.; Lustig, Kevin D.; Friedberg, Ilan; Heppel, Leon A.

doi:10.1016/s0076-6879(89)71046-6

Cited by 14 publications

(6 citation statements)

References 56 publications

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“…In the preliminary experiments, normal rat livers were tested for fuchsin stain ing after infusion of the activator for up to 90 min, but the staining was only weakly and sporadically positive. Simultaneous infusion of Tween 80, a membrane permeabilizing agent (19), evidently increased TBARS release, but the liver became too soft to make slices. Therefore, in the following experiments, livers from PB-pretreated rats were used, in which microsomal Fe 2+ + ADP + NADPH-depend ent lipid peroxidation is reported to be en hanced (20).…”

Section: Infusion Of Adp-fe3+mentioning

confidence: 99%

Histological Detection of Lipid Peroxidation Following Infusion of Tert-Butyl Hydroperoxide and ADP-Iron Complex in Perfused Rat Livers.

Masuda¹,

Yamamori²

1991

Jpn.J.Pharmacol

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ABSTRACT-Lipid peroxidation was assessed histologically and biochemically in hemoglobin-free perfused rat livers using two different types of stimulators. The Schiff reaction of fuchsin with cellular aldehydes was used as a histological index for lipid peroxidation. t-Butyl hydroperoxide (BHP, 0.8 mM) infusion caused a rapid and sus tained release of thiobarbituric acid reactive substances (TBARS) into the effluent perfusate for up to 60 min, which was accompanied by lactate dehydrogenase (LDH) leakage after 30 min. The Schiff positive foci were initially restricted to periportal zones and spread with time to whole areas, accompanied by periportal necrosis. Co infusion of diphenyl-p-phenylenediamine suppressed the TBARS release, with nega tive fuchsin staining, but the LDH leakage was unaffected. Under retrograde perfu sion, BHP produced pericentral staining and necrosis. With 2.5 mM ADP-100'U M Fe3+, little TBARS was released up to 60 min, even though the hepatic TBARS levels increased considerably by this time. By 90 min, marked TBARS release occurred, but LDH leakage remained low. Irrespective of the direction of perfusion, pericentral hepatocytes became Schiff positive after 30 min. The fuchsin staining method may be useful for detecting peroxidized zones of the liver lobules.

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Section: Infusion Of Adp-fe3+mentioning

confidence: 99%

Histological Detection of Lipid Peroxidation Following Infusion of Tert-Butyl Hydroperoxide and ADP-Iron Complex in Perfused Rat Livers.

Masuda¹,

Yamamori²

1991

Jpn.J.Pharmacol

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“…The radioactivity of samples mixed with "Liquiscint" (National Diagnostics, Manville, NJ) was determined by liquid scintillation counting in a Beckman LS 7000 counter. The percent of labeled soluble pool released at each time point was calculated as described by Weisman et al, (1989b). It was verified, by chromatographing samples on polyethyleneimine-cellulose columns as previously described (Magnusson et al, 19761, that a t least 80% of the radioactivity effluxed consisted of adenine nucleotides.…”

Section: Measurement Of Efflux Of Intracellular Soluble Pools Labeledmentioning

confidence: 99%

Permeabilization of transformed mouse fibroblasts by 3′‐O‐(4‐benzoyl)benzoyl adenosine 5′‐triphosphate and the desensitization of the process

González

Ahmed

Lustig

et al. 1989

Journal Cellular Physiology

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A photoreactive analogue of ATP, 3'-O-(4-benzoyl)benzoyl adenosine 5'-triphosphate (BzATP) altered the plasma membrane permeability of transformed 3T6 mouse fibroblasts to normally impermeant molecules as previously reported for ATP, but at lower concentrations. BzATP-induced permeabilization was modulated by pH, temperature, and the ionic composition of the medium similar to the permeabilizing effects of ATP. Conditions known to enhance ATP-induced permeabilization, such as treatment with the mitochondrial uncoupler carbonyl cyanide-m-chlorophenylhydrazone (CCCP) or the Ca2+-calmodulin antagonist trifluoperazine also enhanced BzATP-induced permeabilization. Conditions inhibitory to ATP-induced permeabilization, including chloride replacement or treatment with furosemide or dithiothreitol (DTT), inhibited permeabilization induced by BzATP. The ionic strength of the medium modulated the responsiveness of the cells to ATP and BzATP; a decrease in the ionic strength below isotonicity increased the sensitivity of the cells to the nucleotides, whereas an increase in ionic strength above isotonicity inhibited permeabilization. Prolonged exposure to ATP under non-permeabilizing conditions caused the cells to become insensitive to ATP and BzATP. The densensitization phenomenon provides support for the theory that the permeabilization process is mediated by a receptor for ATP.

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“…Since the pioneering study of Druri and Szent-Gyorgi (1929), a considerable number of publications have shown that these effects are mediated by purinergic receptors (Burnstock and Kennedy, 1985;Burnstock, 1991). Under specific conditions, slightly alkaline buffer and low concentration of bivalent ions, ATP selectively permeabilizes the cell membrane of transformed cells Heppel et al, 1985;Rozengurt et al, 1977;Weisman et al, 1989). Partial permeabilization of macrophages was found under physiological conditions (Greenberg, 1988).…”

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confidence: 99%

Growth inhibition of breast cancer cells induced by exogenous ATP

Spungin

Friedberg

1993

Journal Cellular Physiology

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Addition of ATP (> 0.1 mM) to cultures of human breast cancer T47D cells resulted in an inhibition of cell proliferation. The inhibition was found to be specific for ATP, and dependent on its concentration. Growth inhibition continued for at least three days, although ATP and its hydrolysis products were metabolized within one day. Conditioned medium from ATP-treated cultures (CM+) was found to inhibit the growth of cells that were not exposed to ATP. This is an indication that extracellular factors, besides ATP, are involved in the inhibition process. The inhibition was maintained after dialysis of the CM+, using an 8 kDa cut-off membrane. Conditioned medium from untreated cultures (CM-), however, only slightly affected cell growth. The data suggest that the CM(+)-induced cell growth inhibition is mediated by an ATP-activated growth inhibiting factor. Flow microfluorometry and thymidine incorporation experiments have shown that the growth arrest is mainly due to the elongation of the S-phase of the cell cycle.

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[43] Permeabilizing mammalian cells to macromolecules

Cited by 14 publications

References 56 publications

Histological Detection of Lipid Peroxidation Following Infusion of Tert-Butyl Hydroperoxide and ADP-Iron Complex in Perfused Rat Livers.

Histological Detection of Lipid Peroxidation Following Infusion of Tert-Butyl Hydroperoxide and ADP-Iron Complex in Perfused Rat Livers.

Permeabilization of transformed mouse fibroblasts by 3′‐O‐(4‐benzoyl)benzoyl adenosine 5′‐triphosphate and the desensitization of the process

Growth inhibition of breast cancer cells induced by exogenous ATP

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