Elevation of intracellular cAMP by noradrenaline and vasoactive intestinal peptide in striated ducts isolated from the rabbit mandibular salivary gland.

Salivary gland intralobular ducts are responsible for the modification of the electrolyte composition of the primary fluid secreted by the acini. However, the intracellular messengers that regulate this and other intralobular duct cell processes have not been fully characterized. To investigate the possibility that cAMP-mobilizing agonists may be involved, intralobular (striated) ducts were isolated from the rabbit mandibular salivary gland by tissue dissociation and microdissection and maintained in tissue culture overnight. Individual duct fragments were stimulated with the secretory agonists noradrenaline, vasoactive intestinal peptide (VIP) and substance P and their cAMP content measured by acetylated radioimmunoassay. Both noradrenaline and VIP elevated intracellular cAMP content concentration dependently, but substance P did not. The response to noradrenaline was blocked by the beta-adrenoceptor antagonist propranolol, but not by the alpha-adrenoceptor antagonist prazosin. Application of the VIP analogue [D-p-Cl-Phe6, Leu17]-VIP decreased the VIP-induced cAMP response. These results demonstrate that striated intralobular duct cells possess beta-adrenoceptors and peptidergic receptors that are coupled to adenylate cyclase and activated by noradrenaline and VIP, respectively. By elevating ductal cAMP content, these agonists may regulate both the electrolyte content of the primary saliva and the secretion of protein(s) from the ducts.

Lack of effect of atrial natriuretic peptide and urodilatin on vasopressin-stimulated cyclic AMP production in microdissected rat inner medullary collecting ducts.

It is unclear whether the diuretic effects of atrial natriuretic peptide (ANP) result, in part, from an inhibition of the renal actions of vasopressin. Moreover, accruing evidence suggests that the kidneys themselves may produce an ANP-like peptide, urodilatin, which shares many of the renal actions of ANP. The mechanism underlying the diuretic action of urodilatin has not yet been examined. Accordingly, we have investigated the potential modulatory actions of both ANP and urodilatin on vasopressin-stimulated cyclic AMP (cAMP) production in microdissected inner medullary collecting duct (IMCD) segments of rat kidney. ANP and urodilatin alone (at 10(-8) or 10(-6) M) had no demonstrable effect on cAMP accumulation in IMCD segments. Moreover, neither ANP nor urodilatin (each at 10(-6) M) significantly altered either the profile or the absolute magnitude of the cAMP response stimulated by vasopressin. These findings indicate that neither ANP nor urodilatin interacts with the vasopressin-sensitive adenylate cyclase site in the rat IMCD to contribute to its diuretic actions.

Peptide hormone-stimulated second messenger production in the teleostean nephron.

Renal actions of arginine vasotocin (AVT) and atrial natriuretic peptide (ANP) in teleosts are poorly defined. Administration of these hormones changes renal function, largely explained by systemic vascular effects. The present experiments examine the potential direct action of these hormones on the teleost nephron. This has been assessed from the hormones' abilities to stimulate their second messenger systems within isolated nephroi of the rainbow trout Onchorhynchus mykiss. Following initial experiments to define the experimental conditions, AVT was found to induce a dose-dependent increase in cAMP content of a suspension of nephroi incubated for 10 min at 10 degrees; 10(-5) M AVT provoking an increase of 846% (SW) and 829% (FW) above basal cAMP accumulation of 88 and 125 pmol/mg protein, respectively. This V2-type receptor response to AVT (or AVP) has previously been considered to be present only in the tetrapod kidney. ANP evoked a dose-dependent stimulation of cGMP accumulation from 1.3 (SW) or 1.9 (FW) pmol/mg protein basal levels to 5806 and 4405% of these levels, respectively, at the highest concentration used of 3.3 x 10(-7) M. Further experiments using isolated glomeruli from the eel Anguilla anguilla localized at least a part of the response to ANP to a glomerular site.

Drinking behaviour in sea water and fresh water teleosts, the role of the renin-angiotensin system.

Basal drinking rate and responses to administered angiotensin were examined in 12 species of fish. The responses of representative euryhaline, stenohaline marine and fresh water species to pharmacological manipulation of endogenous renin-angiotensin system (RAS) activity were also investigated.Basal drinking rates were consistently low in stenohaline and euryhaline fresh water fish, and all species examined showed an increased imbibition in response to administered angiotensin. Marine fish drank large volumes of water, rates varying considerably between species, with euryhaline species exhibiting lower rates than stenohaline groups. The extremely high drinking rates observed in the sea scorpion were associated with a high plasma osmolality. With the exception of the sea scorpion, all other species examined in sea water showed a further rise in drinking in response to exogenous angiotensin.Although the freshwater stenohaline carp showed a dipsogenic response to angiotensin, it was apparently unable to evoke this response when fish were acclimated to brackish water. The high drinking rates of both euryhaline and stenohaline fish held in sea water appeared dependent upon an activated endogenous RAS, and were lowered following inhibition of Al to All conversion by Captopril. Drinking was further stimulated in these marine species following stimulation of endogenous RAS activity by the administration of the hypotensive agent Papaverine. The study endorses a role for the RAS in the control of adaptive drinking in euryhaline and stenohaline marine teleosts.

Pituitary and plasma arginine vasotocin levels in teleost fish.

The role of arginine vasotocin (AVT) in teleostean osmoregulation is unclear. Its administration may produce dose-dependent effects on renal fluid loss though the physiological significance of such effects is unknown due to the paucity of data on the normal range in pituitary and plasma AVT levels. The present study addresses these questions by examining the pituitary AVT activity in the flounder and plasma AVT concentrations in the flounder, trout, and carp following alterations in environmental tonicity. Flounder pituitary AVT content, measured by bioassay, was elevated in freshwater (FW)-acclimated compared to seawater (SW)-acclimated fish and rapidly fell or rose, respectively, following transfer from FW to SW or from SW to FW to attain a new steady state. Plasma levels of AVT were higher in FW than in SW in both flounder and trout and it would, therefore, appear that the synthesis, storage, and secretion of AVT may be enhanced in FW. In the stenohaline carp plasma levels of AVT were unchanged on acclimation from FW to 40% SW. In SW flounder there was a significant positive correlation between plasma osmolality and plasma AVT concentration. Such a relationship was not apparent in FW fish, which may reflect differing roles for the hormone in these two extremes of osmotic environment.

The renin-angiotensin system and the regulation of plasma cortisol in the flounder, Platichthys flesus.

The changes in arterial blood pressure and plasma cortisol concentration in response to exogenous angiotensin II (AII) and to manipulation of the endogenous renin-angiotensin system (RAS) have been examined in the flounder, Platichthys flesus. Intravenous [Asp1Val5]AII was pressor in a dose-dependent manner over the range of 0.25-250 micrograms/kg. Sequential blood sampling revealed steroidogenic stimulation, measured as change in plasma cortisol concentration, by doses of AII greater than 2.5 micrograms/kg. Enhanced circulating cortisol levels were measured some time after recovery of normal systemic blood pressure, suggesting that the change in plasma steroid concentration was not entirely dependent on the change in systemic blood pressure. Administration of the vasodilator papaverine produced immediate hypotension followed by gradual recovery in blood pressure, which was accompanied by a sustained increment in circulating cortisol. Both blood pressure recovery and the increased plasma steroid concentration were inhibited by prior treatment with the angiotensin converting enzyme inhibitor captopril. These results are consistent with a physiologically important role for the RAS in the control of plasma cortisol levels which complements its demonstrated pressor and dipsogenic actions in the flounder: the RAS may thus afford an integrating influence on the mechanisms of body fluid homeostasis in this euryhaline species.