Multivalent interaction between asialofetuin and plasma membrane preparations from the rat liver

1980 ◽  
Vol 58 (12) ◽  
pp. 1414-1420 ◽  
Author(s):  
Maria T. Debanne ◽  
Erwin Regoeczi ◽  
Mark W. C. Hatton
Keyword(s):  
Rat Liver ◽  
Plasma Membranes ◽  
Binding Capacity ◽  
Theoretical Models ◽  
Underlying Mechanism ◽  
Marker Enzyme ◽  
Electron Microscopic ◽  
Microscopic Appearance ◽  
Wide Range ◽  
Electron Microscopic Appearance

Binding of bovine asialofetuin by rat liver plasma membranes was studied using different techniques for the separation of the free and bound forms of the glycoprotein and also different approaches to measure nonspecific binding. The membrane preparations had the electron microscopic appearance of a mixture of lamellae and vesicles and their lipid:protein ratios and marker enzyme profiles fell within the range of values available from the literature. The binding capacity was approximately 15 pmol of asialofetuin per milligram of membrane protein.Scatchard plots of the values obtained over a wide range of concentrations (4.8–12.6 μg asialofetuin per 30 μg membrane protein) after incubation at 22 °C showed pronounced non-linearity which, in combination with evaluations according to other theoretical models, was referable to heterogeneity of binding. In sharp contrast, after incubation at 4 °C the Scatchard plot was linear. This difference is interpreted as the expression of a functional, rather than a chemical, heterogeneity in asialofetuin binding. The underlying mechanism is thought to be competition of galactose groups for binding sites with the result that the number of bonds varies between the galactose groups of a bound asialofetuin molecule and the hepatic lectin, depending on the concentration of the glycoprotein in the incubation mixture.

Edward George Gray. 11 January 1924 – 14 August 1999

2002 ◽  
Vol 48 ◽  
pp. 151-165
Author(s):  
R.W. Guillery
Keyword(s):  
Nervous System ◽  
Three Dimensional ◽  
Electron Microscopic ◽  
Early Application ◽  
Microscopic Appearance ◽  
Wide Range ◽  
Neural Tissues ◽  
Synaptic Junctions ◽  
The Central Nervous System ◽  
Electron Microscopic Appearance

George Gray was an early contributor to our knowledge of the electron microscopic appearance of the central nervous system. He was skilful with the difficult techniques for preparing the tissues, worked rapidly, and was an astute observer. Sitting with him in the dark, staring at a dim image that George was moving rapidly as he searched for significant detail, could be an exciting experience. He had clear ideas about features that mattered and could quickly relate the two-dimensional electron microscopic images to the three-dimensional neural structures under investigation. He is best known for his detailed and perceptive description of synaptic junctions in the mammalian neocortex, and his name is still linked to two distinct junctional types (Gray's type 1 and Gray's type 2), now recognized as generally distinguishing excitatory from inhibitory junctions. He studied a wide range of neural tissues, played a significant role in the early isolation of ‘synaptosomes’, contributed greatly to the rapid advance of knowledge that accompanied the early application of the electron microscope to neural tissues, and influenced a great many later fine-structural studies of the nervous system.

Incorporation in vivo of [32P]orthophosphate and [Me-3H]choline into rough microsomal, Golgi, and plasma membranes of rat liver

1977 ◽  
Vol 55 (8) ◽  
pp. 876-885 ◽  
Author(s):  
Patricia L. Chang ◽  
John R. Riordan ◽  
Mario A. Moscarello ◽  
Jennifer M. Sturgess
Keyword(s):  
Endoplasmic Reticulum ◽  
Plasma Membrane ◽  
Rat Liver ◽  
Plasma Membranes ◽  
Specific Activity ◽  
Specific Radioactivity ◽  
Marker Enzyme ◽  
Golgi Membranes ◽  
Endomembrane Flow

To study membrane biogenesis and to test the validity of the endomembrane flow hypothesis, incorporation of 32P and [Me-3H]choline in vivo into membranes of the rat liver was followed. Rough microsomal, Golgi-rich, and plasma membrane fractions were monitored with marker enzyme assays and shown with morphometric analysis to contain 82% rough microsomes, at least 70% Golgi complexes, and 88% plasma membranes, respectively. Membrane subfractions from the rough microsomal and Golgi-rich fractions were prepared by sonic disruption.At 5 to 30 min after 32P injection, the specific radioactivity of phosphatidylcholine was higher in the rough microsomal membranes than in the Golgi membranes. From 1 to 3 h, the specific activity of phosphatidylcholine in Golgi membranes became higher and reached the maximum at about 3 h. Although the plasma membrane had the lowest specific radioactivity throughout 0.25–3 h, it increased rapidly thereafter to attain the highest specific activity at 5 h. Both rough microsomal and plasma membranes reached their maxima at 5 h.The specific radioactivity of [32P]phosphatidylethanolamine in the three membrane fractions was similar to that of [32P]phosphatidylcholine except from 5 to 30 min, when the specific radioactivity of phosphatidylethanolamine in the Golgi membranes was similar to the rough microsomal membranes.At 15 min to 5 h after [Me-3H]choline injection, more than 90% of the radioactivity in all the membranes was acid-precipitable. The specific radioactivities of the acid-precipitated membranes, expressed as dpm per milligram protein, reached the maximum at 3 h. After [Me-3H]choline injection, the specific radioactivity of phosphatidylcholine separated from the lipid extract of the acid-precipitated membranes (dpm per micromole phosphorus) did not differ significantly in the three membrane fractions. The results indicated rapid incorporation of choline into membrane phosphatidylcholine by the rough endoplasmic reticulum, Golgi, and plasma membranes simultaneously.The data with both 32P and [Me-3H]choline precursors did not support the endomembrane flow hypothesis. The Golgi complexes apparently synthesized phosphatidylethanolamine and incorporated choline into phosphatidylcholine as well as the endoplasmic reticulum. The results are discussed with relevance to current hypotheses on the biogenesis and transfer of membrane phospholipids.

scholarly journals REVERSIBLE AND IRREVERSIBLE CHANGES IN THE FINE STRUCTURE OF NERVOUS TISSUE DURING OXYGEN AND GLUCOSE DEPRIVATION

1965 ◽  
Vol 26 (3) ◽  
pp. 885-909 ◽  
Author(s):  
Henry deF. Webster ◽  
Adelbert Ames
Keyword(s):  
Synaptic Vesicles ◽  
Glucose Deprivation ◽  
Control Medium ◽  
Electron Microscopic ◽  
Microscopic Appearance ◽  
Oxygen And Glucose Deprivation ◽  
Golgi Membranes ◽  
Electrophysiological Measurements ◽  
Synaptic Changes ◽  
Electron Microscopic Appearance

Rabbit retinas were fixed for electron microscopy immediately after removing the eye and after incubations in a control medium and in three different deprivation media that were identical with the control except for the omission of glucose, oxygen, or both. A systematic comparison was made of the electron microscopic appearance of the different retinas with particular attention to four regions: rod inner segments, rod synapses, bipolar cell bodies, and ganglion cell myelinated axons. Retinas fixed after 1 hour of incubation in the control medium appeared virtually identical with those fixed immediately after ocular removal. Retinas deprived of oxygen and glucose for only 3 minutes showed generalized swelling of mitochondria and alterations in the structure of the synapses with loss of synaptic vesicles. Extending the combined deprivation caused further mitochondrial swelling and synaptic changes and also led to progressive swelling of the Golgi membranes and the granular endoplasmic reticulum. All these changes were almost completely reversible for up to 20 minutes but were irreversible by 30 minutes, at which time multiple discontinuities had appeared in cell and organelle membranes. Anoxia alone produced alterations similar to those found after somewhat shorter periods of the combined deprivation, whereas glucose withdrawal produced only minor changes. These electron microscopic results correlate quite well with previously reported electrophysiological measurements.

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