Akiyoshi Takahashi

Effects of N-terminal peptide of salmon proopiocortin on interrenal function of the rainbow trout

Effects of N-terminal peptide of salmon proopiocortin (salmon NPP-I) on cortisol secretion was examined in vitro using diced interrenal tissue from the rainbow trout, Salmo gairdneri. ACTH(1-24) at concentrations of 1 to 50 nM stimulated cortisol secretion in dose-dependent manner, whereas salmon NPP-I had no effect over a range of 50 pM to 500 nM. Cortisol secretion in response to various doses of ACTH(1-24) was modified slightly when 1 to 100 nM of salmon NPP-I was added to the incubation medium together with ACTH. An augmentation of in vitro secretion of cortisol in response to ACTH(1-24) was observed when the interrenal was removed from the trout pretreated with one IU of porcine ACTH but not with 10 micrograms of salmon NPP-I. A slight but significant potentiating effect of salmon NPP-I (10 or 100 nM) on the ACTH-induced cortisol secretion was observed when the trout was sensitized to ACTH by porcine ACTH pretreatment. Furthermore, six daily injections of salmon NPP-I into the trout induced hyperplasia of interrenal tissue. These findings suggest that NPP-I, together with ACTH, may be involved in controlling interrenal function in the trout. Such activities could be due to conservative region in the N-terminal portion of NPP.

Plasma profiles of the N-terminal peptide of proopiomelanocortin in the rainbow trout with reference to stress

Plasma levels of the N-terminal peptide of proopiomelanocortin (NPP) were measured in rainbow trout, Salmo gairdneri, following treatment of handling stress with or without administration of dexamethasone, adaptation to white and black background, and maintenance on a constant light/dark cycle. Effects of exogenously administered NPP on plasma constituents were also examined to provide insight into the biological significance of NPP. Thirty minutes of handling stress in shallow water had no effect on plasma levels of NPP during and after the stress period, whereas significant increases in plasma cortisol and glucose were observed. Intraperitoneal administration of dexamethasone blocked the stress-induced elevation of plasma levels of cortisol and caused a depression of plasma NPP. No difference was observed in plasma levels of NPP between trout adapted to a white background and those adapted to a black background. No diurnal changes in NPP were observed under an artificial light/dark cycle (14L/10D light cycle, 0500-1900 hr light) in May and September. Thus, plasma levels of NPP were considerably constant under various physiological conditions, and no synchronism was observed between plasma NPP and cortisol, although NPP modifies the corticotropin-induced release of cortisol from the interrenal. Plasma constituents such as cortisol, total protein, albumin, plasma amino nitrogen, glucose, free fatty acid, ketone body, sodium, potassium, calcium, and magnesium were not altered by intraperitoneal injections of NPP (1 or 10 micrograms) once daily for 6 days (total of six injections) or once every other day for 28 days (14 injections). High concentrations of NPP were found in the plasma 24 hr after cessation of the serial injections of NPP (10 micrograms), suggesting slow metabolic clearance of the peptide.

A radioimmunoassay for N-terminal peptide of chum salmon proopiocortin.

A highly specific radioimmunoassay was developed for N-terminal peptide of salmonid proopiocortin using a guinea pig antiserum to the chum salmon peptide (sNPP 1). Since sNPP I has no tyrosine residue nor free N-terminal amino group, a mixture of minor components of sNPP 1, which have extensions of H-Val-LysGly- and H-Lys-Gly- at the N-terminus, were iodinated by the lactoperoxidase method after incorporation of 3-(phydroxyphenyl)-propionate to the terminal amino groups. Plasma and pituitary extracts of several salmonid species showed parallel displacement to the standard hormone. Samples from carp, goldfish, tilapia, and eel, as well as the plasma of hypophysectomized rainbow trout, showed no crossreactivity. Proopiocortin-related hormones isolated from the chum salmon pituitary, including melanotropins, endorphins, corticotropin-like intermediate lobe peptides, and gonadotropin and prolactin showed negligible cross-reactivity. NPP contents in the pars intermedia of rainbow trout and chum salmon were 10 to 15 times greater than those in the pars distalis. Plasma levels of NPP in the mature chum salmon caught in the bay were about 50ng/ml. Plasma NPP levels in the mature chum salmon of both sexes decreased after transfer from seawater to fresh water. Plasma cortisol showed a concomitant change with NPP, thus supporting previous findings that NPP modulates corticotropin action on the trout interrenal.