scholarly journals THE ISOLATION OF A CELL MEMBRANE FRACTION FROM RAT LIVER

1960 ◽  
Vol 8 (2) ◽  
pp. 413-422 ◽  
Author(s):  
David M. Neville

A procedure is described for isolating cell membranes from rat liver homogenates. 20 gm. of rat liver was homogenized in a Dounce homogenizer in ice cold water buffered to pH 7.5 with NaHCO3, rupturing all of the cells and most nuclei. The diluted homogenate was filtered through cheesecloth to remove precipitated nucleoprotein and centrifuged at 1500 g, 10 minutes, to sediment a crude membrane fraction. The membrane containing sediment was recentrifuged 3 times in conical tubes (1220 g, 10 minutes), the top layer of the 2-layered sediment being retained. Flotation in a sucrose solution d = 1.22 freed the preparation from contaminating cell fragments and nuclear membranes not previously disintegrated. The floating material ∼0.4 ml. was quite homogeneous and consisted of thin amorphous membranes. Electron micrographs revealed numerous double profiles similar in shape and dimensions to apposed liver cell membranes in intact tissue.

1951 ◽  
Vol 34 (5) ◽  
pp. 647-656 ◽  
Author(s):  
Norman G. Anderson ◽  
Karl M. Wilbur

1. The addition of heparin to rat liver, kidney, or brain nuclei has been found to bring about the release of a gel. Chemical analysis and histochemical studies on whole homogenates and isolated nuclei demonstrated that the material released by heparin contained desoxyribonucleic acid (DNA) and protein. The action of heparin on nuclei is interpreted as the result of a combination with the basic proteins of the nucleus with a consequent displacement of DNA. 2. The addition of heparin to a finely divided dilute liver homogenate prepared in a phosphate-sucrose solution at pH 7.1 brings about a marked increase in viscosity which reaches a maximum in 6 to 8 minutes at 23° and then declines. 3. The concentration threshold for the viscosity effect was 0.1 mg. per 100 mg. fresh rat liver, with further increases in viscosity at higher heparin concentrations. Over a period of several hours a marked decrease in response to heparin was observed in homogenates stored at 0°. 4. Fractionation of the homogenate demonstrated that the viscosity increase was due to the presence of the nuclei alone, other components showing no effect. Microscopic observation showed that the increase in viscosity was associated with the appearance of a clear gel around nuclei treated with heparin. 5. Heparin brought about the release of DNA from the nuclei of incubated rat liver, kidney, and brain homogenates. In some instances over half the DNA is found in the supernatant after high speed centrifugation (20 minutes, 21,000 x g). 6. No correlation was found between anticoagulant activity of heparin preparations and their effectiveness in causing an increase in the viscosity of liver homogenates. Desulfated heparin produced none of the results described here for heparin.


1990 ◽  
Vol 167 (3) ◽  
pp. 876-882 ◽  
Author(s):  
Akira Ishikawa ◽  
Ryunosuke Kanamaru ◽  
Akira Wakui ◽  
Shin-ichiro Kanno ◽  
Kenzo Ohtsuki

1954 ◽  
Vol 206 (1) ◽  
pp. 471-481 ◽  
Author(s):  
Ivan D. Frantz ◽  
Nancy L.R. Bucher ◽  
Henny S. Schneider ◽  
Naomi H. McGovern ◽  
Ruth Kingston

1957 ◽  
Vol 225 (2) ◽  
pp. 735-744
Author(s):  
Henry Kamin ◽  
Mildred A. Koon ◽  
Philip Handler
Keyword(s):  

1962 ◽  
Vol 202 (2) ◽  
pp. 343-346 ◽  
Author(s):  
Dennis D. Goetsch ◽  
L. E. McDonald

The effects of glucocorticoid administration on oxygen uptake, glucose and glycogen disappearance, lactic acid formation, and inorganic phosphate and protein levels in rat liver homogenates have been studied. A single injection of hydrocortisone, prednisolone, or 9 α-fluoroprednisolone 5 hr before sacrifice resulted in a highly significant increase in oxygen uptake by rat liver homogenates, whereas chronic administration of prednisolone daily for 7 days caused a marked inhibition in homogenate respiration. Glycolytic rate did not appear to be affected by single injections since endogenous carbohydrate utilization was similar in liver homogenates prepared from control and treated animals. Incubation of liver homogenates under aerobic conditions disclosed that inorganic phosphate levels were decreased in homogenates from corticoid-treated rats, whereas these levels were similar in treated and control liver homogenates incubated under nitrogen. Under anaerobic conditions, liver homogenates from treated rats accumulated lactic acid more rapidly than untreated liver homogenates. Glucocorticoid treatment did not appear to affect protein disappearance since no differences between protein levels in treated and untreated rat liver homogenates were detected following incubation.


1991 ◽  
Vol 266 (7) ◽  
pp. 4322-4328 ◽  
Author(s):  
P Moreau ◽  
M Rodriguez ◽  
C Cassagne ◽  
D M Morré ◽  
D J Morré

Sign in / Sign up

Export Citation Format

Share Document