Positive feedback of hepatic angiotensinogen expression in silver sea bream (Sparus sarba)

Abstract

The renin-angiotensin system (RAS) is involved in the maintenance of fluid homeostasis in vertebrates. Production of the precursor protein, angiotensinogen, is regulated by other components within the RAS. Angiotensin II (Ang II) stimulates the production and secretion of angiotensinogen in many mammalian models. However, the existence of a similar positive feedback mechanism for angiotensinogen has not been demonstrated for any non-mammalian species. In the present study, we have cloned the angiotensinogen for silver sea bream (Sparus sarba) and investigated the role of Ang II on angiotensinogen expression. The nucleotide sequence of angiotensinogen for S. sarba only exhibits a fair resemblance to other fish angiotensinogens and shows 76.6% similarity to that of Takifugu rubripes and 57.2% similarity to that of Danio rerio. Angiotensinogen transcripts have been identified in the brain, liver, kidney, and various parts of the intestine of sea bream, an observation, which probably implies the presence of a local RAS at the tissue level. The liver is probably the major source of angiotensinogen, as it exhibits the highest angiotensinogen transcript abundance among different tissues. Differential angiotensinogen expression was found among different regions of the intestine where the pyloric caeca exhibits the highest expression. Putative Ang I is identified at the N-terminal of the deduced protein with a novel sequence [Asn1, Ile5, His9]-Ang I. Hepatic angiotensinogen expression in sea bream adapted to different salinities remained constant and this is probably due to desensitization of the angiotensin receptors by angiotensin. A positive feedback mechanism of angiotensinogen by Ang II has been demonstrated as exogenous Ang II increased the amount of angiotensinogen transcript in isolated hepatocytes in vitro. Blockade of endogenous RAS by the angiotensin converting enzyme (ACE) inhibitor, captopril, significantly lowered the hepatic expression of angiotensinogen in vivo. The effect of Ang II stimulation on angiotensinogen expression is more potent in fish than that in mammals. These data suggest that the positive feedback mechanism of angiotensinogen by Ang II has already evolved in teleosts and such mechanism may be involved in the maintenance of angiotensinogen secretion under resting and hypertensive conditions.