Analytical Study of Microsomes and Isolated Subcellular Membranes From Rat Liver

Beaufay, Henri; Amar-Costesec, A; Feytmans, Ernest; Thinessempoux, D.; Wibo, Maurice; Robbi, Mariette; Berthet, J

doi:10.1083/jcb.61.1.188

Cited by 745 publications

(241 citation statements)

References 64 publications

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“…The atmosphere in the perfusion chamber was 95% 0 and 5% CO.. After 30 min of equilibration, 0, 0.3, or 0.5 Ag/ml [3 HMCYST-LR was added to the perfusate (119 ml), which was recirculated at a constant flow of 10 ml/min for 60 min. (Cooperstein and Lazarow, 1950) and cytochrome reductase (microsomes, fractions 1,2 and 4) (Beaufay et al, 1974) were measured as previously described.…”

Section: Methodsmentioning

confidence: 99%

Toxicity and kinetics of [3H]microcystin-LR in isolated perfused rat livers

Pace

Robinson

Miura

et al. 1991

Toxicology and Applied Pharmacology

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Section: Methodsmentioning

confidence: 99%

Toxicity and kinetics of [3H]microcystin-LR in isolated perfused rat livers

Pace

Robinson

Miura

et al. 1991

Toxicology and Applied Pharmacology

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“…Alkaline phosphodiesterase I was assayed at pH 9.6 with pnitrophenyl-thymidine 5'-phosphate as substrate. Both assays were done employing the incubation conditions reported for rat liver homogenates by Beaufay et al (11).…”

Section: Methodsmentioning

confidence: 99%

Coordinate Secretion of Acid Hydrolases in Rat Bile

LaRusso¹,

Fowler²

1979

J. Clin. Invest.

102

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A B S T R A C T Three lysosomal glycosidases, j8-glucuronidase (EC 3.2.1.31), ,8-galactosidase (EC 3.2.1.23), and N-acetyl-,8-glucosaminidase (EC 3.2.1.30) have been investigated in bile that was freshly collected from rats through a complete bile fistula. Assay conditions have been established on the basis of appropriate kinetic studies. The biliary excretion patterns for these enzymes were found to vary considerably from rat to rat during the 24-h collection period. In a given animal, however, the three hydrolases were excreted in parallel and showed a gradual increase in activity with time, most marked after 10-12 h of collection. 24-h biliary outputs of the three hydrolases averaged m3% of their respective contents in total liver, and bile diversion had no effect on hepatic glycosidase activity or total protein content. Other enzymes known to be associated primarily with mitochondria, endoplasmic reticulum, and cell sap were also detected in bile, generally in smaller amounts. The biliary excretion of the plasma membrane markers, alkaline phosphodiesterase I and 5'-nucleotidase, however, was comparable to that ofthe lysosomal hydrolases. Biliary excretion of total protein was relatively constant and corresponded to 3.0% ofthe total hepatic protein content per day, whereas biliary bile acid secretion decreased during the first 12 h and then remained constant. Exocytic bulk discharge of hepatocyte lysosomes is proposed as the most likely mechanism for the biliary excretion of lysosomal enzymes. These results call attention to the possible pathophysiologic significance of biliary excretion of hepatic lysosomal contents as a means of residue disposal.A preliminary report of part of this work appeared in 1977.

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“…The NADPH cytochrome c reductase activity was measured from extracts of microsomes as described by Beaufay et al (1974). This method is based on the formation of the reduced form of cytochrome c, cytochrome c red , induced by NADPH, which can be recorded spectrophotochemically at 550 nm.…”

Section: Enzymatic Activity Measurementsmentioning

confidence: 99%

Endoplasmic reticulum and Golgi apparatus are the preferential sites of Foscan® localisation in cultured tumour cells

et al. 2003

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Intracellular photosensitiser localisation significantly influences the mechanism of response to photodynamic therapy (PDT), since the primary sites of damage are closely related to the specific sensitiser distribution. Foscan s subcellular localisation in the MCF-7 human adenocarcinoma cell line has been studied by means of confocal microscopy and microspectrofluorometry. The fluorescence topographic profiles recorded after cells costained with Foscan s and organelle-specific fluorescent probes revealed that Foscan s presents low localisation in lysosomes and a weak accumulation in mitochondria. Alternatively, the Foscan s fluorescence topographic profile turned out to colocalise perfectly with that obtained for the endoplasmic reticulum (ER) and the Golgi apparatus. The patterns of fluorescence derived from confocal microscopy studies were consistent with predominant localisation of Foscan s in these organelles. Furthermore, evaluation of enzymatic activity of selected organelles immediately after laser light irradiation (650 nm) indicated the Golgi apparatus and ER as the primary damaged sites resulting from Foscan s -mediated PDT in the MCF-7 cell line. To our knowledge, this is the first study to demonstrate unambiguously that the ER and the Golgi apparatus are preferential sites of Foscan s accumulation.

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Analytical Study of Microsomes and Isolated Subcellular Membranes From Rat Liver

Cited by 745 publications

References 64 publications

Toxicity and kinetics of [3H]microcystin-LR in isolated perfused rat livers

Toxicity and kinetics of [3H]microcystin-LR in isolated perfused rat livers

Coordinate Secretion of Acid Hydrolases in Rat Bile

Endoplasmic reticulum and Golgi apparatus are the preferential sites of Foscan® localisation in cultured tumour cells

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