Effects of somatostatin on intestinal calcium transport in the rat

The addition of somatostatin (SRIF) to rat descending colon in vitro increased the calcium secretory flux from serosa to mucosa (Js leads to m) and reduced tissue short-circuit current (Isc) but did not alter the absorptive flux from mucosa to serosa (Js leads to m). Js leads to m increased by 37% at 10(-9) M SRIF and by 48% at 10(-6) M. The response to SRIF was not altered by 1,25-dihydroxyvitamin D3 [1,25(OH)2D3], and SRIF did not interfere with stimulation of calcium Jm leads to s by 1,25(OH)2D3. Removal of sodium from the buffer abolished the stimulation of Js leads to m by SRIF without reducing basal Js leads to m. Secretory fluxes of mannitol and calcium were strongly correlated in the presence and absence of SRIF, suggesting that SRIF stimulates a paracellular transepithelial pathway for calcium. In the duodenum, SRIF altered neither calcium Js leads to m nor Isc. In the ileum, calcium Js leads to m increased and Isc decreased, as in the colon, but only by 28 and 12%, respectively. The maximal change in calcium Js leads to m caused by SRIF in these three intestinal segments was negatively correlated with the tissue concentration of immunoreactive SRIF. These results suggest that intestinal calcium secretion could, in part, be regulated by intestinal SRIF.     

Studies on renin release from isolated superfused glomeruli: effects of temperature, urea, ouabain and ethacrynic acid.

1. The effects of different energy substrates, of low temperature, of urea, and of ouabain and ethacrynic acid were studied on the rate of renin release from viable juxtaglomerular cells during superfusion of isolated rat glomeruli. 2. Neither lactate nor glutamate altered renin release rate from that observed using glucose as the sole energy substrate. Succinate 10 mM elevated release transiently but did not influence the release caused by reductions in osmolality through lowering sucrose concentration. 3. Peak renin release was more prolonged and returned more slowly to control following reductions in osmolality in phosphate-Ringer than in bicarbonate-Ringer. 4. At 37 degrees C, the peak of renin released induced by hypo-osmolality was smaller and delayed, and returned earlier to control than at 30 degrees C. Reduction in temperature from 30 to 4 degrees C resulted in a 32-fold increase in basal release rate. At 4 degrees C a 20 m-osmole/kg reduction in tonicity caused an additional 2-5-fold increase in release rate. 6. Increasing superfusate osmolality with urea did not affect basal renin release but 100 mM urea suppressed the releasing effect of a 15 mM reduction in NaCl concentration. 7. Ouabain (10(-4) M) caused a small (33 +/- 9%, P less than 0-025) transient increase in renin release. Ethacrynic acid (10(-3) M) provoked a progressive increase in release reaching 100 +/- 15% above control within 50 min. In the presence of both inhibitors the release provoked by hyposmolality was prolonged. 8. It is concluded that renin release in vitro is a function of actively regulated cell volume and it is proposed that a similar mechanism could underline both barorecptor and macula densa controls of renin secretion in vivo.     

Effect of furosemide and ethacrynic acid on sodium transport and potassium secretion

Like strophanthin K, ethacrynic acid increases the sodium concentration and reduces the potassium concentration in frog urinary bladder tissue, with the result that potassium secretion is reduced; furosemide does not change these concentrations. The results point to differences in the intracellular action of furosemide and ethacrynic acid.Laboratory of Evolution of the Kidney and Water-Salt Metabolism, I. M. Sechenov Institute of Evolutionary Physiology and Biochemistry, Academy of Sciences of the USSR, Leningrad. (Presented by Academician of the Academy of Medical Sciences of the USSR V. N. Chernigovskii.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 9, pp. 319–321, September, 1977.     

Effects of sodium fluxes and ouabain on brain-slice tryptophan transport

Efflux of preloaded [3H]tryptophan from rat cerebral cortex slices has been monitored into superfusion media that were altered in their sodium content. Total replacement of sodium with choline greatly increased the release of tryptophan. This release could be cancelled by re-introducing sodium into the slices. A brief exposure to ouabain, an efficient inhibitor of Na+, K+-ATPase activity, only slightly increased tryptophan efflux at the concentration of 0.1 mM, whereas at 1.0 mM it produced a similar effect as the sodium-free medium. Accordingly, when the slices were superfused in the presence of ouabain and sodium, the change of medium to sodium-free caused much greater relative enhancement of tryptophan efflux with 0.1 than 1.0 mM ouabain. Tryptophan efflux was modified by changes in sodium fluxes also in slices initially depleted of sodium ions and treated with ouabain. The results suggest that the sodium-free medium and ouabain have a similar mechanism of action in modifying the tryptophan transport, and that the cation gradients across the cell membranes are more crucial for normal amino acid transport than the functional Na+, K+-ATPase.     

A comparison of the effects of ouabain and ethacrynic acid on the dog kidney in vivo and in vitro

Summary The effects of ouabain, ethacrynic acid andn-butyl-biguanide, alone or in combination, on the dog kidney have been studied by infusion of the drugs into the renal artery in vivo, with simultaneous determinations of the clearances of creatinine and PAH, renal blood flow, oxygen consumption, and reabsorptions of sodium, potassium, chloride and glucose; after removal of the organ, the tissue water and intracellular ion contents, the oxygen consumption and transport capacity for glycine, -methylglucoside and PAH in cortical slices, and the activities of Na⁺–K⁺-ATPase in microsomal fractions from cortical homogenates were determined.The most characteristic action of ouabain and ethacrynic acid was the induction of considerable patriuresis, but their effects were not additive. Ethacrynic acid provoked a pronounced kaliuresis, which was partially reversed when ouabain was administered simultaneously. Chloride reabsorption was inhibited by ethacrynic acid more than by ouabain. Only ouabain induced glucosuria.Ethacrynic acid increased the renal blood flow and reduced both the glomerular filtration rate and the oxygen consumption in vivo. Ouabain had little effect on these parameters.The degree of hydration of cortical chunks was augmented by both ouabain and ethacrynic acid. Ouabain increased intracellular sodium and reduced potassium, whereas ethacrynic acid primarily affected the levels of sodium and chloride. In this respect, the effects of the two drugs were additive. Only ouabain irreversibly inhibited the Na⁺–K⁺-ATPase of the renal cortex, and only ouabain exerted an inhibitory action on sugar and amino-acid accumulation during a subsequent incubation of tissue slices in vitro.It is concluded that ouabain and ethacrynic acid have different modes of action, though it is unclear why in vivo little additivity of their effects on sodium reabsorption can be detected.     

Vitamin D-dependent, particulate calcium-binding activity and intestinal calcium transport.

A vitamin D-dependent calcium-binding activity of relatively high molecular weight has been identified in the particulate fraction of rat small intestinal mucosa. The Ca-binding activity is sedimented at 140,000 X g after treatment of the mucosal particulate fraction with Triton X-114. Intestinal brush-border suspensions can also be used as starting material. The Ca-binding component is inactivated by heat and repeated freeze-thawing and consists of one or more protein complexes in the range of 0.5-1.0 million mol wt as indicated by gel filtration. The Ca-binding activity correlates positively with known features of the intestinal Ca transport mechanism, as demonstrated by studies of the distribution in the small intestine and the effects of vitamin D, dietary Ca, cycloheximide treatment, and rat age. It is suggested that the component might function in the transit of Ca across the brush-border surface to the cytosol of intestinal mucosal cells.     

Relation between cell na extrusion and transtubular absorption in the perfused toad kidney: the effect of K,ouabain and ethacrynic acid

Summary Transtubular absorption of Na and Cl, and intracellular ion concentrations were evaluated in toad kidneys perfused with solutions containing K and without K, and in the presence of 1 mM Ouabain and 1 mM Ethacrynic acid. The following values were obtained with 8.5 mM K: Transtubular absorption of Na and Cl68% (percent of filtered load); cell content 294 mole Na, 433 mole K, 100 mole Cl/g solids. Lack of K in the perfusate diminished transtubular absorption to 25% and the cells gain 244 mole Na/g solids, and lose an equimolecular quantity of K. The process is reversible upon raising the K concentration in the perfusate. Ouabain inhibits transtubular absorption to 6%; the cells lose about 110 mole K/g solids, but cellular Na is maintained at the control levels. Ethacrynic acid inhibits transtubular absorption to 3%; the cells approximately double their Na and Cl content, but their K is maintained at the control levels. These observations cannot be explained exclusively in terms of an effect on the distal tubule. Probably proximal as well as distal tubules are involved. A single Na pump seems insufficient to account for all experimental findings. The existence of two separate pumps is therefore proposed.     

Calcium ATPase and intestinal calcium transport in uremic rats

Calcium ATPase, an enzyme involved in intestinal calcium transport, was measured in homogenates of duodenal mucosal scrapings of normal and uremic rats. The effects of calcium deprivation and treatment with 1 alpha,25-dihydroxycholecalciferol [1,25-(OH)2D3] were investigated as well. Uremia decreased the enzyme activity and impaired the rise after calcium deprivation as observed in intact rats. The 1,25-(OH)2D3 treatment increased the enzyme activity in uremic animals and resulted in an identical response to calcium deprivation as observed in intact rats; parathyroidectomy abolished this effect. A striking correlation between everted duodenal gut sac calcium transport and calcium ATPase activity could be demonstrated for all groups of rats studied. It is concluded that the calcium ATPase activity is linked to the production of 1,25-(OH)2D3 as well as to an additional factor, probably parathyroid hormone. The close relationship between enzyme activity and in vitro calcium transport, even during constant physiological supplementation with 1,25-(OH)2D3, suggests an autonomous role of the calcium ATPase activity for mediation of calcium transport in the duodenum in addition to the well-known mechanisms related to vitamin D and its metabolites.     

Effects of ouabain on potassium transport in the perfused bullfrog kidney.

We studied the effect of ouabain on transepithelial and cellular potassium transport in the isolated perfused bullfrog kidney. We used recently developed techniques for estimating the unidirectional reabsorptive and secretory K fluxes (Jr and Js) and measuring the kinetics of cellular K transport. Two hours of perfusion with 1 X 10(-6) M ouabain did not affect GFR, reduced fractional Na reabsorption 57%, increased K excretion 41%, and inhibited Jr 34%. Js rose 68% at 60 min and then returned to the control level. Ninety minutes of perfusion with 5 X 10(-6) M ouabain reduced GFR 28% and fractional Na reabsorption 76%. K excretion rapidly increased 71% within 30 min and then fell to 60% of the control level, while Jr fell 64%. Js rose 42% in 30 min and then fell to 23% of the control level. Both doses reduced K uptake into cellular pools from the circulation and increased the rate coefficients for efflux into tubular fluid. The data indicate that ouabain inhibited reabsorption and transiently accelerated the rate of loss of K from the cells into the tubular fluid. This initially masked the ultimate inhibition of K secretion from the circulation into the tubular fluid.     

Effects of ureteral occlusion and ethacrynic acid infusion on renal prostaglandin degradation in the dog

The two major renal prostaglandins PGE2 and PGI2 are partly metabolized during a single passage of the kidney. To examine whether stopping glomerular filtration affected the renal degradation, PGE2 and PGI2 were infused into the suprarenal aorta of dogs during ureteral occlusion. Prostaglandin synthesis was blocked by indomethacin, 10 mg kg-1 b.w. i.v. About 20% of PGI2 and 80-90% of PGE2 were metabolized during one passage through the kidney. Prostaglandin degradation and arterial input were proportional (r greater than 0.95). Compared to control conditions at free urine flow, PGI2 degradation was not changed, whereas the degradation of PGE2 was slightly increased by ureteral occlusion. Ethacrynic acid might reduce degradation of PGE2 by inhibiting two degradation enzymes. To examine the influence of ethacrynic acid, PGE2 was infused in different doses into the suprarenal aorta of dogs before and after administration of ethacrynic acid 3 mg kg-1 b.w. i.v. At all dose levels of PGE2, 75-80% was degraded by one passage through the kidney, whether ethacrynic acid was administered or not. However, although ethacrynic acid did not alter the total renal output, the urinary fraction was reduced from 20-30% to 10-15%. We conclude that degradation of both PGE2 and PGI2 is mainly confined to the blood vessels, and that ethacrynic acid in conventional doses does not prevent degradation of PGE2, but redistributes PGE2 output from urine to renal venous blood.