The effect of lipid extraction on electron-microscopic images of plant membranes

1973 ◽  
Vol 51 (6) ◽  
pp. 1221-1229 ◽  
Author(s):  
Elizabeth S. Swanson ◽  
William W. Thomson ◽  
J. Brian Mudd
Keyword(s):  
Structural Integrity ◽  
Lipid Extraction ◽  
Glutaraldehyde Fixation ◽  
Specific Lipids ◽  
Electron Microscopic ◽  
Plant Membranes ◽  
Acetone Extracts ◽  
Layer Chromatography ◽  
Density Pattern

An evaluation was made of the role of lipids in electron-microscopic membrane images of plant cells by comparing extracted lipids with changes in the ultrastructural membrane images. Lipids were extracted from tobacco leaves with a series of acetone concentrations. In a parallel series, glutaraldehyde fixation preceded lipid extraction. Thin-layer chromatography of the acetone extracts showed no major difference in the lipids extracted with and without glutaraldehyde fixation, but different concentrations of acetone removed specific lipids. Electron micrographs of tissues not previously fixed with glutaraldehyde showed a disruption of all membrane images at acetone concentrations greater than 30%. From these studies it appears that lipids are involved in the formation of electron-microscopic membrane images, but to a different degree in the various membranes. The general form of mitochondria and chloroplast grana was not dependent upon lipid, though lipid was required for the typical density pattern of the granal partitions. The bounding membranes of the mitochondria and chloroplasts were lost with extraction of galactolipids, sulfolipids, and phospholipids. The plasmalemma, tonoplast, and microbody membranes lost their typical density pattern and their structural integrity with the extraction of phospholipids.

scholarly journals THE ULTRASTRUCTURE OF LIPID-DEPLETED ROD PHOTORECEPTOR MEMBRANES

1974 ◽  
Vol 63 (2) ◽  
pp. 587-598 ◽  
Author(s):  
Izhak Nir ◽  
Michael O. Hall
Keyword(s):  
Lipid Extraction ◽  
Fatty Acid Analysis ◽  
Major Protein ◽  
Rod Photoreceptor ◽  
Thin Sections ◽  
Retinal Rod ◽  
Layer Chromatography ◽  
Chloroform Methanol ◽  
Photoreceptor Membranes

The structure of lipid-depleted retinal rod photoreceptor membranes was studied by means of electron microscopy. Aldehyde-fixed retinas were exhaustively extracted with acetone, chloroform-methanol, and acidified chloroform-methanol. The effect of prefixation on the extractability of lipids was evaluated by means of thin-layer chromatography and fatty acid analysis. Prefixation with glutaraldehyde rendered 38% of the phospholipids unextractable, while only 7% were unextractable after formaldehyde fixation. Embedding the retina in a lipid-retaining, polymerizable glutaraldehyde-urea mixture allows a comparison of the interaction of OsO4 with lipid-depleted membranes and rod disk membranes which contain all their lipids. A decrease in electron density and a deterioration of membrane fine structure in lipid-depleted tissue are correlated with the extent of lipid extraction. These observations are indicative of the role of the lipid bilayer in the ultrastructural visualization of membrane structure with OsO4. Negatively stained thin sections of extracted tissue reveal substructures in the lipid-depleted rod membranes. These substructures are probably the opsin molecules which are the major protein component of retinal rod photoreceptor membranes.

Stannous fluoride treatment of acid-etched caries-like lesions of enamel: A scanning electron microscopic study

1984 ◽  
Vol 42 ◽  
pp. 720-721
Author(s):  
M. John Hicks ◽  
Leon M. Silverstone ◽  
David G. Gantt ◽  
Catherine M. Flaitz
Keyword(s):  
Acid Etching ◽  
Surface Zone ◽  
Electron Microscopic ◽  
Fluoride Treatment ◽  
Stannous Fluoride ◽  
Scanning Electron Microscopic Study ◽  
Demineralized Enamel ◽  
Intact Surface ◽  
Topical Fluoride

Although fluoride levels become elevated in sound enamel following a topical fluoride treatment, the caries-preventive effect of fluoride is thought to be due primarily to the role of fluoride in remineralization of clinically undetectable enamel lesions and hypomineralized enamel. During lesion formation, redistribution of fluoride from the enamel surface to the subsurface demineralized enamel occurs. This results in a surface zone with a relatively low fluoride content. In order to maintain an intact surface zone over a carious lesion, it may be necessary to replenish the fluoride levels with an exogenous fluoride source. By acid-etching the lesion surface, a more reactive surface is made available for fluoride interaction. In addition, porosities and etching patterns may be created, allowing for bonding of a caries-resistant resin material to the lesion surface. The purpose of this study was to determine the integrity of the caries-like lesion surface following acid-etching and subsequent stannous fluoride treatment (SnF2).

Scanning electron microscopic study of collagen fibrillogenesis and extracellular compartmentalization in the chick embryo tendon

1986 ◽  
Vol 44 ◽  
pp. 208-209
Author(s):  
Grace C.H. Yang
Keyword(s):  
Chick Embryo ◽  
Extracellular Space ◽  
Fibroblast Cell ◽  
Collagen Fibrils ◽  
Electron Microscopic ◽  
Voltage Electron ◽  
Scanning Electron Microscopic Study ◽  
Microscopic Studies ◽  
And Function

The size and organization of collagen fibrils in the extracellular matrix is an important determinant of tissue structure and function. The synthesis and deposition of collagen involves multiple steps which begin within the cell and continue in the extracellular space. High-voltage electron microscopic studies of the chick embryo cornea and tendon suggested that the extracellular space is compartmentalized by the fibroblasts for the regulation of collagen fibril, bundle, and tissue specific macroaggregate formation. The purpose of this study is to gather direct evidence regarding the association of the fibroblast cell surface with newly formed collagen fibrils, and to define the role of the fibroblast in the control and the precise positioning of collagen fibrils, bundles, and macroaggregates during chick tendon development.

scholarly journals Lipid Transporters Beam Signals from Cell Membranes

Membranes ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 562
Author(s):  
Miliça Ristovski ◽  
Danny Farhat ◽  
Shelly Ellaine M. Bancud ◽  
Jyh-Yeuan Lee
Keyword(s):  
Membrane Proteins ◽  
Lipid Bilayers ◽  
Lipid Composition ◽  
Structural Integrity ◽  
Current Knowledge ◽  
Integral Membrane Proteins ◽  
Cellular Membranes ◽  
Cytoplasmic Proteins ◽  
Lipid Transporters

Lipid composition in cellular membranes plays an important role in maintaining the structural integrity of cells and in regulating cellular signaling that controls functions of both membrane-anchored and cytoplasmic proteins. ATP-dependent ABC and P4-ATPase lipid transporters, two integral membrane proteins, are known to contribute to lipid translocation across the lipid bilayers on the cellular membranes. In this review, we will highlight current knowledge about the role of cholesterol and phospholipids of cellular membranes in regulating cell signaling and how lipid transporters participate this process.

The effect of oestrogen withdrawal on cardiac Ca2+ regulation and the influence of GPER1

2020 ◽  
Vol 41 (Supplement_2) ◽  
Author(s):  
A.J Francis ◽  
J.M Firth ◽  
N Islam ◽  
J Gorelik ◽  
K.T MacLeod
Keyword(s):  
Heart Failure ◽  
Structural Integrity ◽  
Left Ventricular ◽  
Funding Source ◽  
Hormonal Changes ◽  
Ion Conductance ◽  
Developing Heart ◽  
Rapid Stimulation ◽  
Post Menopausal

Abstract Background Post-menopausal women have an enhanced risk of developing heart failure, attributed to declining oestrogen levels during menopause. However, the signalling mechanisms remain undetermined. Purpose We aim to determine the role of G-protein coupled oestrogenic receptor 1 (GPER1) in intracellular Ca2+ regulation and the consequences of hormonal changes that may exacerbate the pathophysiology of heart failure. Methods Ovariectomy (OVx) (mimics menopausal hormone changes) or sham surgeries were conducted on female guinea pigs. Left ventricular cardiomyocytes were isolated 150-days post-operatively for experimental use. Cellular t-tubule network and structural integrity was measured using fluorescent di-8-ANEPPs staining and scanning ion conductance microscopy. GPER1 expression and localisation was measured by Western blot and immunostaining. The role of GPER1 activation was measured using selective agonist G-1 in electrophysiological and Ca2+-sensitive dye fluorescence experiments. Results Following oestrogen withdrawal, the t-tubule network density decreased by 13% and z-groove index reduced by 15%. GPER1 predominantly localised to the peri-nuclear endoplasmic reticulum and its expression increased by 32% in OVx. Action potential duration (APD) prolonged in OVx and following GPER1 activation, APD90 shortened by 11% and 25% in sham and OVx respectively. OVx cells had larger peak inward Ca2+ current (ICaL) (by 22%) and sarcoplasmic reticulum (SR) Ca2+ content (by 13%), compared with sham. While GPER1 activation had little effect on peak ICaL or SR content, it reduced Ca2+ transient amplitude (by 20%), SR fractional release (by 11%) in OVx cells. The frequency of occurrence of spontaneous Ca2+ waves evoked by periods of rapid stimulation reduced by 40% and wave-free survival time prolonged in OVx cells following GPER1 activation. Conclusions In the hearts of an animal species whose electrophysiology and intracellular Ca2+ regulation is akin to humans, we show that following oestrogen deficiency, the t-tubule network is down-regulated and becomes disorganised, GPER1 expression is increased and its activation induces negative inotropic responses in cardiomyocytes. This may limit the adverse changes to Ca2+ signalling reported in OVx that could be pro-arrhythmic and exacerbate the progression to heart failure. Funding Acknowledgement Type of funding source: Foundation. Main funding source(s): British Heart Foundation

Etude ultrastructurale des relations hôte–parasite au cours de l'infection des pommes par le Phytophthora cactorum

1981 ◽  
Vol 59 (2) ◽  
pp. 251-263 ◽  
Author(s):  
X. Mourichon ◽  
G. Sallé
Keyword(s):  
Cell Walls ◽  
Susceptible Variety ◽  
Host Cells ◽  
Phytophthora Cactorum ◽  
Electron Microscopic ◽  
Susceptible Host ◽  
Golden Delicious ◽  
Extrahaustorial Matrix ◽  
Apple Fruits

An electron microscopic study was performed on haustoria of Phytophthora cactorum (L. et C.) Schroeter developed in tissues of two cultivars of apple fruits: a susceptible variety ('Golden delicious') and a resistant one ('Belle de Boskoop'). Ultrastructure of intercellular hyphae and some aspects of their penetration between contiguous host cells were described. A light dissolution of the host cell walls was observed. Ontogenic investigations indicated that in the susceptible host, the wall of the fungal haustoria was covered with a dense-stained extrahaustorial matrix. Its origin and its polysaccharide nature were demonstrated. On the other hand, the resistant host developed, immediately after the inoculation, a papilla which gave rise, later on, to a sheath enclosing adult haustoria. The role of these callosic structures in the phenomenon of resistance was discussed.

A High-Throughput Liposome Substrate Assay with Automated Lipid Extraction Process for PI 3-Kinase

2008 ◽  
Vol 13 (9) ◽  
pp. 906-911 ◽  
Author(s):  
Trupti Lingaraj ◽  
John Donovan ◽  
Zhi Li ◽  
Ping Li ◽  
Amanda Doucette ◽  
...  
Keyword(s):  
High Throughput ◽  
Lipid Extraction ◽  
Extraction Process ◽  
Enzyme Mechanism ◽  
Class I ◽  
Lipid Kinase ◽  
Chromatography Analysis ◽  
Regulate Cell Growth ◽  
Lipid Kinases ◽  
Layer Chromatography

The signaling pathways involving lipid kinase class I phosphatidylinositol 3-kinases (PI 3-kinases) regulate cell growth, proliferation, and survival. Class I PI 3-kinases catalyze the conversion of PI (4,5)P2 to PI (3,4,5)P3, which acts as a lipid second messenger to activate mitogenic signaling cascades. Recently, p110α, a class IA PI 3-kinase, was found to be mutated frequently in many human cancers. Therefore, it is increasingly studied as an anticancer drug target. Traditionally, PI 3-kinase activities have been studied using liposome substrates. This method, however, is hampered significantly by the labor-intensive manual lipid extraction followed by a low-throughput thin-layer chromatography analysis. The authors describe a high-throughput liposome substrate-based assay based on an automated lipid extraction method that allows them to study PI 3-kinase enzyme mechanism and quantitatively measure inhibitor activity using liposome substrates in a high-throughput mode. This improved assay format can easily be extended to study other classes of phosphoinositide lipid kinases. ( Journal of Biomolecular Screening 2008:906-911)

scholarly journals Role of the Ndc1 interaction network in yeast nuclear pore complex assembly and maintenance

2009 ◽  
Vol 185 (3) ◽  
pp. 475-491 ◽  
Author(s):  
Evgeny Onischenko ◽  
Leslie H. Stanton ◽  
Alexis S. Madrid ◽  
Thomas Kieselbach ◽  
Karsten Weis
Keyword(s):  
Nuclear Pore Complex ◽  
Structural Integrity ◽  
Fluorescent Protein ◽  
Interaction Network ◽  
Nucleocytoplasmic Transport ◽  
Nuclear Pore ◽  
Binding Partners ◽  
Interaction Partners ◽  
Redundant Functions

The nuclear pore complex (NPC) mediates all nucleocytoplasmic transport, yet its structure and biogenesis remain poorly understood. In this study, we have functionally characterized interaction partners of the yeast transmembrane nucleoporin Ndc1. Ndc1 forms a distinct complex with the transmembrane proteins Pom152 and Pom34 and two alternative complexes with the soluble nucleoporins Nup53 and Nup59, which in turn bind to Nup170 and Nup157. The transmembrane and soluble Ndc1-binding partners have redundant functions at the NPC, and disruption of both groups of interactions causes defects in Ndc1 targeting and in NPC structure accompanied by significant pore dilation. Using photoconvertible fluorescent protein fusions, we further show that the depletion of Pom34 in cells that lack NUP53 and NUP59 blocks new NPC assembly and leads to the reversible accumulation of newly made nucleoporins in cytoplasmic foci. Therefore, Ndc1 together with its interaction partners are collectively essential for the biosynthesis and structural integrity of yeast NPCs.

Mass spectrometry as a versatile ancillary technique for the rapid in situ identification of lichen metabolites directly from TLC plates

2017 ◽  
Vol 49 (5) ◽  
pp. 507-520 ◽  
Author(s):  
Pierre LE POGAM ◽  
Aline PILLOT ◽  
Françoise LOHEZIC-LE DEVEHAT ◽  
Anne-Cécile LE LAMER ◽  
Béatrice LEGOUIN ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Accurate Identification ◽  
Chemical Profiling ◽  
Analytical Strategies ◽  
Tlc Plates ◽  
Lichen Metabolites ◽  
Acetone Extracts ◽  
Layer Chromatography ◽  
Structural Classes

AbstractThin-layer chromatography (TLC) still enjoys widespread popularity among lichenologists as one of the fastest and simplest analytical strategies, today remaining the primary method of assessing the secondary product content of lichens. The pitfalls associated with this approach are well known as TLC leads to characterizing compounds by comparison with standards rather than properly identifying them, which might lead to erroneous assignments, accounting for the long-held interest in hyphenating TLC with dedicated identification tools. As such, commercially available TLC/Mass Spectrometry (MS) interfaces can be easily connected to any brand of mass spectrometer without adjustments. The spots of interest are extracted from the TLC plate to retrieve mass spectrometric signals within one minute, thereby ensuring accurate identification of the chromatographed substances. The results of this hyphenated strategy for lichens are presented here by 1) describing the TLC migration and direct MS analysis of single lichen metabolites of various structural classes, 2) highlighting it through the chemical profiling of crude acetone extracts of a set of lichens of known chemical composition, and finally 3) applying it to a lichen of unknown profile, Usnea trachycarpa.

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