Action of Saponin on Biological Cell Membranes

Glauert, Audrey M.; Dingle, J. T.; Lucy, J. A.

doi:10.1038/196953a0

Cited by 273 publications

(136 citation statements)

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“…Also, diverse saponins such as β-sitosterol (XVI) present in Azadirachta indica A., andrographolide (XVII), a diterpenoid lactone, obtained from Andrographis paniculata Nees and gymnemic acid IV (XVIII) isolated from Gymnema sylvestre R., are agents of glycemic control in animal models [130,131]. The roles of bioactive principles in bringing glycemic control to bear in both experimental animals and humans have been exhaustively described elsewhere [6,114] Adjuvant activities of saponins from traditional Chinese medicinal herbs such as Panax ginseng, Astragalus species, Panax notoginseng, Cochinchina momordica, Glycyrrhiza uralensis and Achyranthes bidentata have been previously described [132]. The immune-stimulatory activities of Quil A (consists of a mixture of related saponins) isolated from the bark of Quillaja (a tree), which is used as an adjuvant in selected veterinary vaccines has been acknowledged [113].…”

Section: Saponinsmentioning

confidence: 99%

Medicinal Potentials and Toxicity Concerns of Bioactive Principles

CO¹

2015

Med Aromat Plants

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Section: Saponinsmentioning

confidence: 99%

Medicinal Potentials and Toxicity Concerns of Bioactive Principles

CO¹

2015

Med Aromat Plants

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“…We describe an extraction protocol in which saponin, a mild detergent selective for cholesterol (15,16) Cell Extraction and Fixation. Subconfluent cells were pretreated with taxol (5 pug/ml), washed once in phosphatebuffered saline (PBS), and extracted at 40C with cytoskeleton buffer (CSK; 10 mM Pipes, pH 6.8/100 mM NaCI/300 mM sucrose/3 mM MgCl2/2 mM EGTA/4 mM vanadyl riboside complex/1.2 mM phenylmethylsulfonyl fluoride) containing either 0.01% (vol/vol) saponin (Sigma) for 7 min or 0.5% (vol/vol) Triton X-100 for 1 min.…”

Section: Introductionmentioning

confidence: 99%

“…This simple but powerful technique is well suited to imaging threedimensional structures, which, although invisible in conventional micrographs, afford sharp, well-defined embedmentfree images (7-9). The unembedded cytoskeleton and nuclear matrix have proven remarkably rigid and can even be cut into ultrathin sections without excessive deformation (10-14).We describe an extraction protocol in which saponin, a mild detergent selective for cholesterol (15,16) Cell Extraction and Fixation. Subconfluent cells were pretreated with taxol (5 pug/ml), washed once in phosphatebuffered saline (PBS), and extracted at 40C with cytoskeleton buffer (CSK; 10 mM Pipes, pH 6.8/100 mM NaCI/300 mM sucrose/3 mM MgCl2/2 mM EGTA/4 mM vanadyl riboside complex/1.2 mM phenylmethylsulfonyl fluoride) containing either 0.01% (vol/vol) saponin (Sigma) for 7 min or 0.5% (vol/vol) Triton X-100 for 1 min.…”

mentioning

confidence: 99%

Imaging cytoskeleton--mitochondrial membrane attachments by embedment-free electron microscopy of saponin-extracted cells.

Lin

Krockmalnic

Penman

1990

Proc. Natl. Acad. Sci. U.S.A.

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Embedment-free electron microscopy images the cytoskeleton and nuclear matrix, which are very difficult to visualize in conventional electron micrographs. However, to be effective, cell structures must be depleted of soluble proteins, which otherwise shroud cell architecture. Nonionic detergents effect this extraction, releasing soluble proteins but also destroying all membranes. Saponin can permeabilize plasma membranes, releasing soluble proteins while preserving many cytoplasmic membranes. Stereoscopic electron microscopy of resinless sections shows the many connections of the cytoskeleton to mitochondrial membranes.The complex morphologies and motilities ofmany cytoplasmic organelles likely reflect tethering to the cytoskeleton. However, little is known of interactions between the cytoskeleton and cytoplasmic membranes since conventional electron microscopy reveals little of their nature. While micrographs of Epon-embedded, ultrathin sections show membranes clearly, the protein networks of the cytoskeleton are invisible. Conversely, embedment-free electron microscopy affords sharp, three-dimensional images of the cytoskeleton of detergentextracted cells but so far has not been useful for cell membranes. We show here a method for visualizing cytoplasmic membranes and their connections to the cytoskeleton.The conventional electron microscope thin section was originally developed expressly for examining cell membranes (1). The similar electron-scattering cross-sections of embedding plastic and specimen preclude image formation. Heavy metal atoms, adhered to the section surface, delineate the specimen but only where it emerges from the plastic. Membranes, with osmium as a mordant for binding metal atoms, intersect the section surface, giving clear, meaningful images. Cytoskeleton fibers, for the most part, do not.In embedment-free electron microscopy, specimens are imaged directly in vacuo (2-4). Freed of embedding plastic, biological elements yield high contrast images with no need for staining. However, the embedment-free specimen must be depleted of soluble proteins, which otherwise shroud the cytoskeleton, confounding images to near uselessness. This is most easily done by extraction with a nonionic detergent, such as Triton X-100, in a physiological buffer (5-7). Membranes are solubilized and soluble proteins simply diff-use away. This simple but powerful technique is well suited to imaging threedimensional structures, which, although invisible in conventional micrographs, afford sharp, well-defined embedmentfree images (7-9). The unembedded cytoskeleton and nuclear matrix have proven remarkably rigid and can even be cut into ultrathin sections without excessive deformation (10-14).We describe an extraction protocol in which saponin, a mild detergent selective for cholesterol (15,16) Cell Extraction and Fixation. Subconfluent cells were pretreated with taxol (5 pug/ml), washed once in phosphatebuffered saline (PBS), and extracted at 40C with cytoskeleton buffer (CSK; 10 mM Pipes, pH 6.8/100 mM NaCI/...

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“…At high concentrations, saponin produces a pore in the cell membrane as a result of the formation of insoluble complexes with cholesterol (Bangham & Home, 1962;Glauert et al, 1962;Ohtsuki & Ozawa, 1977;Ohtsuki et al, 1978). For this reason some saponins are used as a tool to make chemically skinned preparations of cardiac muscles and smooth muscles (Endo & Kitazawa, 1978;Saida & Nonomura, 1978).…”

Section: Introductionmentioning

confidence: 99%

Positive inotropic action of saponins on isolated atrial and papillary muscles from the guinea‐pig

Enomoto

Ito

Kawagoe

et al. 1986

British J Pharmacology

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The effects of several saponins of animal and plant origin on the contractile activity of atrial and papillary muscles of the guinea‐pig were tested. In a concentration of 1 × 10−5 M, holothurin‐A (HL‐A), holothurin‐B, echinoside‐A, echinoside‐B and sakuraso‐saponin (Saku) exhibited positive inotropic and chronotropic actions whereas desacyl‐jego‐saponin and ginsenoside‐Rd did not. Saponins having a positive inotropic action caused haemolysis of rabbit erythrocytes whereas those without inotropic action did not cause haemolysis. The positive inotropic action of saponins was not affected by practolol, chlorpheniramine, cimetidine and indomethacin. Verapamil (10−6 M) inhibited the inotropic actions due to HL‐A and isoprenaline (10−8 M) to the same extent but had a small efect on those due to ouabain (10−7 M). In high K+ (30 mM K+) medium where the action potential and the contraction were depressed, HL‐A, Saku and isoprenaline restored the action potential and the contraction of the ‘slow response’ type whereas ouabain failed to do so. In normal medium HL‐A and Saku reduced the resting membrane potential by 15–20 mV. These results suggest that modification of the Ca channel is involved in the positive inotropic action of saponins.

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Action of Saponin on Biological Cell Membranes

Cited by 273 publications

References 3 publications

Medicinal Potentials and Toxicity Concerns of Bioactive Principles

Medicinal Potentials and Toxicity Concerns of Bioactive Principles

Imaging cytoskeleton--mitochondrial membrane attachments by embedment-free electron microscopy of saponin-extracted cells.

Positive inotropic action of saponins on isolated atrial and papillary muscles from the guinea‐pig

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