Circulating Inhibitor of Sodium—Potassium-Activated Adenosine Triphosphatase after Expansion of Extracellular Fluid Volume in Rats

1977 ◽  
Vol 53 (4) ◽  
pp. 329-334 ◽  
Author(s):  
H. C. Gonick ◽  
H. J. Kramer ◽  
W. Paul ◽  
E. Lu
Keyword(s):  
Inhibitory Activity ◽  
Adenosine Triphosphatase ◽  
Rat Kidney ◽  
Gel Filtration ◽  
Extracellular Fluid ◽  
Weight Fraction ◽  
Sodium Potassium ◽  
Lysine Vasopressin ◽  
Control Serum ◽  
Isotonic Sodium Chloride

1. Serum was collected from normal rats and from rats volume-expanded with isotonic sodium chloride solution. 2. The serum was fractionated by gel filtration on Sephadex G-25 and each fraction was tested for inhibitory activity against sodium—potassium-activated adenosine triphosphatase prepared from rat kidney homogenate. 3. A single low-molecular-weight fraction, eluting after the salts and after exogenously added lysine-vasopressin, had significantly greater enzyme inhibitory activity when obtained from serum of volume-expanded animals than from control serum. 4. As this fraction has been shown in previous independent studies to contain a natriuretic factor, it may be concluded that one property of this factor is the ability to inhibit sodium—potassium-activated adenosine triphosphatase.

Basal Activity of the Natriuretic Factor Extracted from the Rat Kidney as a Function of the Diet and its Role in the Regulation of the Acute Sodium Balance

1980 ◽  
Vol 58 (5) ◽  
pp. 385-391 ◽  
Author(s):  
F. Louis ◽  
H. Favre
Keyword(s):  
Sodium Intake ◽  
Rat Kidney ◽  
Extracellular Volume ◽  
Gel Filtration ◽  
Kidney Tissue ◽  
Weight Fraction ◽  
Sodium Content ◽  
Sodium Balance ◽  
Low Molecular Weight ◽  
Extracellular Volume Expansion

1. The effect of the sodium content of the diet on the natriuretic activity of an extract from the kidneys was studied in non-expanded and volume-expanded rats. 2. The kidney tissue was homogenized and the supernatant fractionated by gel filtration on Sephadex G-25. A single low-molecular-weight fraction eluted after the salt possessed the natriuretic activity and was tested on a rat bioassay. 3. The natriuretic activity of the fraction obtained from the kidneys of non-expanded rats was related to the sodium intake. 4. After an acute extracellular volume expansion, the natriuretic activity obtained from the fraction extracted from the kidneys was much greater than before expansion and was related to the dietary intake of sodium.

Renal handling of gentamicin by the Munich-Wistar rat

1981 ◽  
Vol 241 (6) ◽  
pp. F645-F648
Author(s):  
A. U. Sheth ◽  
H. O. Senekjian ◽  
H. Babino ◽  
T. F. Knight ◽  
E. J. Weinman
Keyword(s):  
Rat Kidney ◽  
Extracellular Fluid ◽  
Distal Tubule ◽  
Wistar Rat ◽  
Significant Degree ◽  
Proximal Tubules ◽  
Loop Of Henle ◽  
Renal Handling ◽  
Isotonic Sodium Chloride ◽  
Nephron Heterogeneity

Free-flow micropuncture studies were performed in the Munich-Wistar rat to determine the nephron sites of gentamicin transport. The rats were infused with radiolabeled gentamicin and were volume-expanded with either isotonic sodium chloride or sodium bicarbonate. The fractional delivery of ultrafilterable gentamicin (FD) out of the superficial proximal tubule averaged 75% in both groups of animals. FD to the superficial distal tubule averaged 58.1 +/- 2.1% in saline-infused and 91.6 +/- 5.3% in bicarbonate-infused animals. FD to Henle's loop was 118.2 +/- 8.8 and 124.3 +/- 8.1% in saline- and bicarbonate-infused animals, respectively. FD to the base and tip of the papilla was not significantly different between the two groups and averaged 97.3 +/- 2.9 and 96.1 +/- 3.0% in the saline-infused and 100.5 +/- 2.6 and 94.3 +/- 2.8% in the bicarbonate-infused animals. FD to the base of the papilla was significantly lower than that to the loop of Henle in both groups. These studies indicate that gentamicin undergoes net reabsorption in juxtamedullary proximal tubules. Expansion of the extracellular fluid volume with saline or bicarbonate results in differing rates of gentamicin delivery to the superficial distal tubule but not to the base of the papilla. These findings suggest a significant degree of nephron heterogeneity for gentamicin transport in the rat kidney.

INACTIVATION OF ARGININE- AND LYSINE-VASOPRESSIN BY SLICES FROM DIFFERENT ZONES OF THE RAT KIDNEY AND BY RAT LIVER SLICES

1963 ◽  
Vol 44 (4) ◽  
pp. 545-562 ◽  
Author(s):  
N. A. Thorn ◽  
N. B. S. Willumsen
Keyword(s):  
Antidiuretic Hormone ◽  
Rat Kidney ◽  
Extracellular Fluid ◽  
Outer Cortex ◽  
Liver Slices ◽  
Lysine Vasopressin ◽  
Site Of Action ◽  
The Difference ◽  
High Concentrations ◽  
Inner Cortex

ABSTRACT A method was developed for studying the inactivation of antidiuretic hormone in slices from different zones of the rat kidney. The essential features of the method are: The use of thin slices from 4 zones: outer cortex, inner cortex, outer medulla and papilla which are incubated aerobically with the hormone in a medium that has a composition essentially the same as rat extracellular fluid, including a calcium concentration of 2.5 mmol. The inactivation of arginine- and lysine-vasopressin by these slices at 25° C and 37° C for 2 hours was studied. All zones inactivated both vasopressins, but the most marked inactivation activity per mg dry tissue was found in the papillary zone. High concentrations of sodium chloride in the incubation medium did not affect the inactivation of vasopressin by papillary slices. The amounts of arginine- and lysine-vasopressin (expressed in rat pressor units) which were inactivated per mg dry tissue in the different zones during the incubation period were equivalent. Similar results were found in experiments on the inactivation of the two hormones by liver slices. These findings are discussed in relation to the problems of the site of action of antidiuretic hormone and the difference in the antidiuretic response to arginine- and lysine-vasopressin which is found after i. v. injection in the rat.

scholarly journals On the Molecular Characterization of the Sodium-Potassium Transport Adenosine Triphosphatase

1969 ◽  
Vol 54 (1) ◽  
pp. 327-342 ◽  
Author(s):  
Lowell E. Hokin
Keyword(s):  
Adenosine Triphosphatase ◽  
Gel Filtration ◽  
Coupled Transport ◽  
Sodium Potassium ◽  
Cardiotonic Steroid ◽  
Nonionic Detergent ◽  
Last Stage ◽  
Derivatives Of

The phosphorylated intermediate in the (Na + K)-activated adenosine triphosphatase (Na-K ATPase) has been characterized as an L-glutamyl-γ-phosphate residue in the enzyme. This has been accomplished by digestion of the phosphorylated and nonphosphorylated forms of the enzyme with pepsin, reaction of the pepsin digests with [2,3-3H]N-(n-propyl)hydroxylmine, further digestion of the derivatized peptides with pronase in the presence of carrier L-glutamyl-γ-N-(n-propyl)hydroxamate and carrier L-aspartyl-N-(n-propyl)hydroxamate, and chromatographic purification. An increment in radioactivity migrated with authentic L-glutamyl-γ-N-(n-propyl)hydroxamate in a total of seven electrophoretic and chromatographic systems and on gel filtration. No increment in radioactivity was associated with authentic L-aspartyl-ß-N-(n-propyl)hydroxamate in five out of the seven chromatographic and electrophoretic systems. At the last stage of purification the radioactivity from the phosphorylated enzyme which migrated as L-glutamyl-γ-N-(n-propyl)hydroxamate was 2½ times that from the nonphosphorylated enzyme. On the basis of these results it is concluded that the phosphorylated intermediate in the Na-K ATPase is an L-glutamyl-γ-phosphate residue. The beef brain Na-K ATPase has been solubilized with the nonionic detergent, Lubrol, and has been purified 10 times over that in the original microsomes. The soluble enzyme remains stable in the presence of ATP and either Na+ or K+. If the partially purified enzyme is electrophoresed in 3% polyacrylamide, followed by incubation with ATP, Na+, K+, and Mg++, a single, somewhat diffuse, ATPase band, which is ouabain-sensitive is seen. Protein impurities are also seen on the gel. Gel electrophoresis, after treatment of the partially purified enzyme with phenol-acetic acid-urea, shows about 12 discrete protein bands. Studies on the site-directed alkylation of the (Na + K)-activated adenosine triphosphatase with haloacetate derivatives of cardiotonic steroids are reviewed. Efforts are now underway to specifically alkylate the cardiotonic steroid site of the Na-K ATPase with hellebrigenin 3-[2-3H]iodoacetate and to purify the subunit of the enzyme containing the cardiotonic steroid site by following radioactivity. Finally, a working model for the role of the Na-K ATPase in the coupled transport of Na and K is presented.

In Vivo Studies on the Inhibition of Coagulation by Fractionated Heparin and by a Heparin Analogue I. Effects of Heparin Fractions

1981 ◽  
Vol 46 (03) ◽  
pp. 612-616 ◽  
Author(s):  
U Schmitz-Huebner ◽  
L Balleisen ◽  
F Asbeck ◽  
J van de Loo
Keyword(s):  
Molecular Weight ◽  
Day Treatment ◽  
Gel Filtration ◽  
Weight Fraction ◽  
In Vivo Studies ◽  
Low Molecular Weight ◽  
High Molecular Weight Fraction ◽  
Platelet Factor ◽  
Heparin Fractions

SummaryHigh and low molecular weight heparin fractions obtained by gel filtration chromatography of sodium mucosal heparin were injected subcutaneously into six healthy volunteers and compared with the unfractionated substance in a cross-over trial. Equal doses of 5,000 U were administered twice daily over a period of three days and heparin activity was repeatedly controlled before and 2, 4, 8 hrs after injection by means of the APTT, the anti-Xa clotting test and a chromogenic substrate assay. In addition, the in vivo effect of subcutaneously administered fractionated heparin on platelet function was examined on three of the volunteers. The results show that s.c. injections of the low molecular weight fraction induced markedly higher anti-Xa activity than injections of the other preparations. At the same time, APTT results did not significantly differ. Unfractionated heparin and the high molecular weight fraction enhanced ADP-induced platelet aggregation and collagen-mediated MDA production, while the low molecular weight fraction hardly affected these assays, but potently inhibited thrombin-induced MDA production. All heparin preparations stimulated the release of platelet Factor 4 in plasma. During the three-day treatment periods, no side-effects and no significant changes in the response to heparin injections were detected.

Mechanism of Inhibitory Action on Platelet Activation of a Phospholipase A2 Isolated from Lachesis muta (Bushmaster) Snake Venom

1997 ◽  
Vol 78 (05) ◽  
pp. 1372-1380 ◽  
Author(s):  
André L Fuly ◽  
Olga L T Machado ◽  
Elias W Alves ◽  
Célia R Carlinis
Keyword(s):  
Platelet Aggregation ◽  
Enzymatic Activity ◽  
Phospholipase A2 ◽  
Inhibitory Activity ◽  
Thermal Inactivation ◽  
Anticoagulant Activity ◽  
Gel Filtration ◽  
Apparent Molecular Weight ◽  
Crude Venom ◽  
Lachesis Muta

SummaryCrude venom from Lachesis muta exhibited procoagulant, proteolytic and phospholipase A2 activities. A phospholipase A2, denoted LM-PLA2 was purified from L. muta venom to homogeneity, through a combination of chromatographic steps involving gel-filtration on Sephacryl S-200 HR and reverse phase chromatography on a C2/C18 column. LM-PLA2 presented a single polypeptide chain with an isoelectric point at pH 4.7 and apparent molecular weight of 17 kDa. Partial aminoacid sequence indicated a high degree of homology for LM-PLA2 with other PLA2 from different sources.LM-PLA2 displayed a potent enzymatic activity as measured by indirect hemolysis of red blood cells but it was neither lethal when injected i.p. into mice nor did it present anticoagulant activity. Furthermore, LM-PLA2 displayed a moderate inhibitory activity on the aggregation of rabbit platelets induced by low levels of ADP, thrombin and arachidonate. In contrast, platelet aggregation induced by high doses of collagen was strongly inhibited by LM-PLA2 as well as ATP-release. Treatment of the protein with p-bromophenacyl bromide or 2-mercapto-ethanol, as well as thermal inactivation studies, suggested that the platelet inhibitory effect of LM-PLA2 is dependent on its enzymatic activity. Thus, the platelet inhibitory activity of LM-PLA2 was shown to be dependent on the hydrolysis of plasma phospholipids and/or lipoproteins, most probably those rich in phosphatidylcholine. Surprisingly, lyso-phosphatidylcholine released by LM-PLA2 from plasma was shown to preferentially inhibited collagen-induced platelet aggregation, in contrast to other PLA2s, whose plasma hydrolytic products indistinctly affect platelet’s response to several agonists.

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