Release of an antidiuretic substance by bradykinin in the rat

1. Water-loaded rats under ethanol anaesthesia were used to determine whether bradykinin releases ADH and whether the release is a reflex response to hypotension or the result of direct stimulation of the central nervous system by the peptide.2. Intravenous injections of bradykinin caused hypotension followed by prolonged antidiuresis; both responses were dose dependent.3. The antidiuresis caused by bradykinin could not be accounted for by hypotension alone. Haemorrhage and intravenous injections of isoprenaline, which caused falls in arterial blood pressure similar to those caused by bradykinin, produced only short-lasting antidiuretic responses.4. Intracarotid injections of bradykinin caused antidiuresis with little or no preceding hypotension. No antidiuresis was produced by intracarotid injections of isoprenaline.5. After intravenous injection of bradykinin, small amounts of ADH were identified in whole blood and, on a few occasions only, in concentrated urine. However, ADH was not detected in blood extracts.6. After intravenous injection of nicotine in doses which caused antidiureses similar to that caused by bradykinin, ADH was regularly detected in large amounts in blood extracts and in unconcentrated urine.7. Bradykinin did not cause prolonged antidiureses when injected into a rat with congenital diabetes insipidus.8. ADH release was not increased when isolated rat neural lobes were incubated with bradykinin although release was elevated by an increase in the K(+) concentration of the incubation medium.9. It is suggested that bradykinin causes release of an antidiuretic substance by a direct action on the central nervous system. The possibility that this substance is ADH is discussed.     

Influence of mucosal and serosal pH on antidiuretic action in frog urinary bladder.

Mucosal acidification to pH 6.5 reduced by 88% the oxytocin- (2.2 x 10(-8) M) elicited increase of water permeability in frog urinary bladder. Mucosal alkalinization (pH 10.5) increased by as much as 200% the response to the same concentration of oxytocin. These effects were not observed when supramaximal concentrations of oxytocin were imployed. Similar changes were found when the serosal pH was modified. The hydrosmotic responses elicited by serosal hypertonicity or cyclic AMP plus theophylline were also affected by mucosal or serosal changes of the hydrogen in concentration, suggesting an effect at a post-cyclic AMP level. Important interactions were found between luminal pH and serosal hypertonicity when experimental conditions were employed similar to those observed in the collecting duct of mammalian nephron. Freeze-fracture studies showed that the number of intramembranous aggregates of particles induced by ADH in the luminal membrane was reduced by mucosal acidification and augmented by an increase in medium pH.     

Effect of verapamil on the hydroosmotic response to antidiuretic hormone in toad urinary bladder

To investigate the role of the calcium ion in the hydroosmotic response to antidiuretic hormone (ADH), the effects of verapamil, an inhibitor of calcium ion entry into cells, on stimulated water flow was examined in vitro in the toad urinary bladder. Verapamil, 50 micro M, decreased ADH-stimulated osmotic water flow from 23.4 +/- 4.1 to 9.9 +/- 3.3 mg . min-1 . hemibladder-1 (mean +/- SE, n = 12, P < 0.001). That this inhibitory effect was due to a verapamil-induced alteration in cellular calcium metabolism is suggested by the findings that 45Ca2+ uptake by isolated toad bladder epithelial cells was reduced nearly 50% in the presence of verapamil and that reversibility of verapamil's inhibitory action was calcium dependent. Additionally, verapamil reduced theophylline- (20 mM) stimulated water flow from 22.8 +/- 2.7 to 9.5 +/- 2.9 mg . min-1 . hemibladder-1 (n = 7, P < 0.001) but enhanced cAMP- (10 mM) induced water flow from 12.8 +/- 2.5 to 21.6 +/- 1.1 ng . min-1 . hemibladder-1 (n = 7, P < 0.001). The latter effect was due, at least in part, to a direct inhibitory effect by verapamil on phosphodiesterase activity of toad bladder homogenates. These results, therefore, suggest that the calcium ion is an important coupling factor at the level of the adenylate cyclase enzyme complex for the stimulus-reabsorption coupling between ADH and the transporting epithelia of the toad urinary bladder.     

Effects of various ambient temperatures and of heating and cooling the hypothalamus and cervical spinal cord on antidiuretic hormone secretion and urinary osmolality in pigs.

1. Plasma ADH concentration, urinary and plasma osmolality and haematocrit were measured in young pigs placed in cold, thermoneutral, warm and hot ambient temperatures. In some experiments a thermode placed in the hypothalamus or over the cervical spinal cord was heated or cooled at various ambient temperatures. 2. Plasma ADH concentration remained at a low level (0-5--5 muu. ml.-1) over 2 hr or 3 hr periods when the pigs were in cold, thermoneutral or warm ambient temperatures. A hot environment, which caused a marked rise in the pigs' rectal temperature, was associated with a large rise in plasma ADH level. 3. The rise in plasma ADH level which occurred during an increase in body temperature was consistently and completely suppressed by simultaneous cooling of the thermode in the pre-optic region to 5 to 10 degrees C. When the thermode was in the region of the supraoptic nucleus the rise in ADH was only partly suppressed, and when it was over the cervical cord it was only sometimes suppressed. 4. Cooling the thermodes in any position at a cold or thermoneutral ambient temperature, or heating them at a thermoneutral or warm ambient temperature, caused no consistent change in ADH. 5. A diuresis, with a urinary flow-rate of at most 1 ml. min-1 and minimal urinary osmolality of 53 m-osmole kg-1, was observed on only three occasions, twice during cooling of a thermode in the hypothalamus and once after the end of a period when the thermode was heated. In each case, the plasma ADH was less than 2 muu. ml.-1. 6. A slight rise of haematocrit in cold ambient conditions and a slight fall in the warm were observed. Otherwise changes in haematocrit were trivial, and a shift of water between vascular system and interstitium could not be invoked to account for changes in ADH levels. Observed variation of plasma osmolality was also slight.     

Ca2+ entry through the apical membrane reduces antidiuretic hormone-induced hydroosmotic response in toad urinary bladder

The role of Ca²⁺ in the regulation of antidiuretic hormone(ADH)-induced water permeability of the apical membrane of the toad urinary bladder was examined. The effects of modifying Ca²⁺ entry through the apical membrane of toad urinary bladders on the hydroosmotic water flow _H2O) and short circuit current (I _sc) were measured. In most experiments the bladders were treated with small amounts of Ag⁺ (10^–7 mol/l) on the apical side. This treatment was used because previous experiments indicate that it markedly increases alkali-earth cation fluxes through an amiloride-insensitive cation channel in the apical membrane of the urinary bladder. Moreover, when Ca²⁺ is the major cation in the apical solution of these Ag⁺-treated bladders, I _sc is mostly due to Ca²⁺ entry through the apical membrane. Ag⁺ increased I _sc and simultaneously inhibited _H2O in bladders perfused with Ca²⁺ solutions on the apical side. Addition of La³⁺ to the apical solution reversed the stimulation of I _sc and the inhibition of _H2O produced by Ag⁺. When bladders were perfused with Ca²⁺-free solutions on the apical side, addition of Ag⁺ did not inhibit _H2O while the stimulation of cation movements through the amiloride-insensitive cation channel persisted. In bladders perfused with apical Ca²⁺ solutions and treated with chlorophenyl thio-cyclic adenosine monophosphate (ClPheS-cAMP) the addition of Ag⁺ did not inhibit _H2O while it still increased I _sc. Finally, addition of Ca²⁺ to the apical solution of bladders not treated with Ag⁺ reduced _H2O. These results taken together with other findings in the literature suggest: (1) Ca²⁺ entry through the Ag⁺-treated amiloride-insensitive cation channel of the apical membrane inhibits _H2O; (2) the effects of Ca²⁺ entry are at a regulatory site that precedes the interaction of cAMP with the water channels; (3) it is also possible that Ca²⁺ entry through the unmodified amiloride-insensitive cation channel may have some inhibitory effect on _H2O.