Pituitary proopiomelanocortin-derived peptides and hypothalamus-pituitary-interrenal axis activity in gilthead sea bream (Sparus aurata) during prolonged crowding stress: differential regulation of adrenocorticotropin hormone and alpha-melanocyte-stimulating hormone release by corticotropin-releasing hormone and thyrotropin-releasing hormone

Plasma levels of cortisol, growth hormone (GH), adrenocorticotropin hormone (ACTH), alpha-melanocyte-stimulating hormone (alpha-MSH), N-acetyl-beta-endorphin, in vitro ACTH-stimulated cortisol secretion, and in vitro corticotropin-releasing hormone (CRH)- and thyrotropin-releasing hormone (TRH)-stimulated ACTH and alpha-MSH secretion were investigated in gilthead sea bream exposed to high stocking density (30 kg m(-3)) for 23 days. Within 3 days after the onset of crowding, plasma levels of cortisol, ACTH, alpha-MSH, and N-acetyl-beta-endorphin were above control values. After 7 days, plasma parameters had returned to control levels, but at 23 days, cortisol, alpha-MSH, and N-acetyl-beta-endorphin levels were again elevated over controls, indicating a long-term activation of the melanotrope cells. In contrast, crowding stress elicited a prolonged reduction in plasma GH levels concomitant with the increased hypothalamus-pituitary-interrenal axis (HPI) activation. Crowding stress enhanced cortisol secretory activity of the unstimulated interrenal cells. However, interrenal tissue from crowded fish in vitro displayed an attenuated response to ACTH stimulation compared with tissue from control fish, indicating a desensitization of these cells to ACTH during crowding. The involvement of pituitary proopiomelanocortin-derived peptides in the HPI axis of sea bream is indicated by the observed modulation of the CRH and TRH responsiveness of the corticotropes and melanotropes in crowded fish. At day 1, when there were crowding-induced plasma increases in ACTH and alpha-MSH, there was an attenuated CRH-stimulated but not TRH-stimulated, ACTH release. However, at that time, CRH- and TRH-induced responses of alpha-MSH secretion, and the unstimulated secretory activity of the MSH cells, were enhanced in crowded sea bream. These data provide evidence for stimulatory roles of multiple hypothalamic (CRH and TRH) and pituitary (ACTH and alpha-MSH) peptides in the activation of the hypothalamus-pituitary-interrenal axis under crowding conditions in sea bream.     

Dynamics of background adaptation in Xenopus laevis: role of catecholamines and melanophore-stimulating hormone

The pars intermedia of the pituitary gland in Xenopus laevis secretes alpha-melanophore-stimulating hormone (alpha-MSH), which causes dispersion of pigment in dermal melanophores in animals on a black background. In the present study we have determined plasma levels of alpha-MSH in animals undergoing adaptation to white and black backgrounds. Plasma values of black-adapted animals were high and decreased rapidly after transfer to a white background, as did the degree of pigment dispersion in dermal melanophores. Plasma MSH values of white-adapted animals were below the detection limit of our radioimmunoassay. Transfer of white animals to a black background resulted in complete dispersion of melanophore pigment within a few hours, but plasma MSH levels remained low for at least 24 hr. This discrepancy between plasma MSH and degree of pigment dispersion suggested the involvement of an additional factor for stimulating dispersion. Results of in vitro and in vivo experiments with receptor agonists and antagonists indicated that a beta-adrenergic mechanism, functioning at the level of the melanophore, is involved in the stimulation of pigment dispersion during the early stages of background adaptation.     

N-terminal acetylation of melanophore-stimulating hormone in the pars intermedia of Xenopus laevis is a physiologically regulated process

The N-terminal acetylation of melanophore-stimulating hormone (MSH) increases the melanotropic potency of the peptide. This modification may be important in amphibians, where MSH causes skin darkening during adaptation to black background. This study examines the acetylation status of the peptide in the toad Xenopus laevis under different conditions of background adaptation. Acetylated and nonacetylated alpha-MSH were analyzed by high-performance liquid chromatography and quantified by radioimmunoassay. The acetylation status of alpha-MSH was analyzed in tissue, in plasma and in media obtained from in vitro incubation of neurointermediate lobe tissue. Nonacetylated MSH is the major form of alpha-MSH in tissue from both black- and white-background-adapted animals. In plasma of black-adapted animals only acetylated alpha-MSH could be detected. Plasma MSH levels of white-adapted animals were barely detectable. Analysis of peptides secreted during in vitro incubations of neurointermediate lobe tissue from black-adapted animals showed that the relative contribution of alpha-MSH to the immunoreactive profile was considerably enhanced, which supports the concept that acetylation of MSH in Xenopus is associated with the secretory process. Acetylation capacity of tissue from white-adapted animals was much lower and only after several days on black background was full capacity acquired. It is suggested that de novo biosynthesis of acetylation enzymes may be necessary for the acquisition of the acetylation capacity. Transfer of black animals to white background caused a rapid decrease in acetylation capacity, which suggests that factors involved in the rapid inhibition of secretion might also regulate acetylation.(ABSTRACT TRUNCATED AT 250 WORDS)     

Localisation and identification of melanocyte-stimulating hormones in the fish brain

The existence of melanocyte-stimulating hormone (MSH) in fish brains was investigated by a range of techniques: radioimmunoassay, HPLC, bioassay, and immunocytochemistry. Immunoreactive alpha MSH (ir alpha MSH) was detected by radioimmunoassay in all regions of carp and trout brains, with the highest concentration in the basal hypothalamus. In trout, ir alpha MSH cell bodies were located by immunocytochemistry only periventricularly, in the medial basal hypothalamus near the third ventricle, whereas in the carp ir alpha MSH staining was seen both in periventricular cells and also in some of the magnocellular neurones in the lateral hypothalamus. When white-adapted fish were transferred to a black tank for 6 days, the melanin-concentrating hormone (MCH) content of the basal hypothalamus of both carp and trout increased 2- and 4.6-fold, respectively, but the alpha MSH content did not change in either species. Analysis by HPLC of pituitary gland, hypothalamic, and optic tectal extracts revealed that the pituitary contains desacetyl, monoacetyl, and diacetyl alpha MSH, although the ratio of these forms differed in the two species. The hypothalamus and optic tectum, however, contained predominantly the desacetyl form of alpha MSH. Bioassays for MSH in the HPLC fractions revealed the existence of presumptive beta MSH in both the pituitary and hypothalamus. An argument is advanced that the periventricular ir alpha MSH neurones are homologous with the proopiomelanocortin cells of the arcuate nucleus in mammals, and that the immunocytochemical alpha MSH-like activity in the MCH neurones may not be authentic alpha MSH.     

Pituitary and interrenal function in gilthead sea bream (Sparus aurata L., Teleostei) after handling and confinement stress

Dynamics of adrenocorticotropin (ACTH), alpha melanocyte-stimulating hormone (alpha-MSH), N-acetylated-beta-endorphin (N-ac-beta-END), cortisol, and growth hormone (GH) were investigated in gilthead sea bream (Sparus aurata) stressed by handling plus confinement. As indices of the secondary stress response, plasma levels of glucose, lactate, and plasma ions were monitored. Within 1 h, plasma cortisol and ACTH levels increased above the control values but GH levels decreased. Subsequently, at 24 h cortisol and ACTH levels had declined, but were still higher than in controls, whereas GH levels had recovered after 4 h. Regarding the melanotrope peptides, there were no differences in plasma levels of alpha-MSH and N-ac-beta-END, but pituitary stores of these peptides were severely depleted already after 1 h, as were ACTH stores. Pituitary contents of proopiomelanocortin (POMC)-derived hormones did not show significant differences from 72 h onward. Therefore, the results indicate that both handling and confinement affected the corticotropes of the pars distalis and the melanotropes of the neurointermediate lobe but at different magnitudes. The possible involvement of corticotropin-releasing hormone (CRH) in the regulation of pituitary POMC-producing cell types under these conditions was indicated by the in vitro dose-dependent effect of the peptide on release of ACTH, alpha-MSH, and N-ac-beta-END. The corticocotropes appeared more responsive, and approximately 10-fold more sensitive, to CRH compared with the melanotropes. The ACTH-releasing potency of 1 nM CRH was inhibited 75% following pretreatment of the whole pituitary gland with 400 nM of the CRH antagonist alpha-helical CRH(9-41).     

The stress response of the gilthead sea bream (Sparus aurata L.) to air exposure and confinement.

We investigated short-term effects (up to 24 h) of air exposure and confinement, and long-term effects (up to 11 days) of confinement, to elucidate signalling pathways in the stress response of gilthead sea bream Sparus aurata L. Plasma glucose and lactate were taken as indicators of sympathetic activation, and alpha-melanocyte stimulating hormone (alpha-MSH), adrenocorticotrophic hormone (ACTH) and cortisol as indicators of activation of the brain-pituitary-interrenal (BPI) axis. Air exposure for 3 min resulted, within 30 min, in an increase in plasma concentrations of cortisol, alpha-MSH, glucose, lactate, osmolality and plasma Na, Cl and Mg. Plasma ACTH and beta-endorphin and plasma K, Ca and P did not change. We conclude that air exposure mainly activates the brain-sympathetic-chromaffin cell (BSC) axis. In fish confined at a density of 70 kg/m(3) (compared with 4 kg/m(3) in controls), cortisol, ACTH and alpha-MSH increased within 1 h, indicating activation of the BPI axis. Plasma glucose, Na, Cl and Mg increased with an 8 h delay compared with the response to air exposure. No changes in plasma lactate, osmolality, K, Ca and P were observed. Long-term confinement induced a biphasic cortisol response with peaks at 1 h and at 2 and 3 days. A gradual increase in plasma beta-endorphin concentrations peaked at 7 days; the concentration of alpha-MSH increased rapidly within 1 h and then declined to control values 4 days after the onset of confinement. No changes in ACTH were detected. Our data provide evidence that a stressor-specific activation of the BSC and BPI axes may occur in Sparus aurata.     

Effects of background adaptation on the pituitary and plasma concentrations of some pro-opiomelanocortin-related peptides in the rainbow trout (Salmo gairdneri)

Radioimmunoassays for alpha-MSH, beta-MSH, ACTH and endorphin were used to measure pituitary concentrations of these peptides in rainbow trout during adaptation to black and white backgrounds. There was no difference in the pituitary content of any of these peptides between long-term black- and white-adapted trout. Plasma levels of alpha-MSH immunoreactivity were significantly higher in black-adapted trout than in white-adapted trout. Time-course studies revealed that although the body colour of trout showed an initial rapid adaptation to background colour, this was not paralleled by a corresponding change in plasma alpha-MSH levels. These only showed significant changes after 7 or more days of background adaptation, when melanophore recruitment or degradation occurred on black or white backgrounds respectively. Intravenous administration of mammalian alpha-MSH, salmon beta-MSH I or antibodies to these peptides did not affect short-term background adaptation. However, long-term administration of mammalian alpha-MSH via osmotic minipump maintained melanophore numbers in grey-adapted trout transferred to a white background, although this observation was based on only two fish. It is concluded that peptides derived from pro-opiomelanocortin do not appear to be involved in controlling physiological colour change but may be involved in regulating morphological colour change of the rainbow trout.     

Changes in pituitary and plasma levels of MSH in teleosts during physiological colour change

Immunoreactive αMSH was measured in the plasma of eels, trout, and flounder, following different periods of adaptation to illuminated white or black backgrounds. In both eels and trout, plasma hormone titres changed rapidly in response to the background colour. By contrast, in the flounder, whose pituitary melanotrophs appeared cytologically unresponsive to background change, plasma hormone levels remained similar for fish from black or white tanks following a 3-week adaptation period, but a difference became apparent after nearly 7 weeks adaptation. Similarly, bioactive and immunoreactive levels of MSH in trout pituitary glands usually showed significant changes with black ground colour, whereas flounder pituitary levels were unaltered. It is concluded that MSH is involved in physiological colour changes in eels and trout, but probably not in flounders.     

Acetylation of melanocyte-stimulating hormone and beta-endorphin in the pars intermedia of the perinatal pituitary gland in the mouse

This report concerns ontogenetic aspects of the production and in vitro release of NH2-terminally acetylated forms of melanocyte-stimulating hormone (alpha-MSH) and beta-endorphin by the pars intermedia of the pituitary gland of the mouse. In vitro biosynthetic analysis and radioimmunoassay revealed that approximately 12 h before birth most of the MSH in the fetal pars intermedia is present as des-N alpha-acetyl alpha-MSH. The same non-acetylated peptide is at this stage also the major release form of melanotropin. In 1-day-old mice the level of alpha-MSH and diacetylated alpha-MSH had increased considerably, although des-N alpha-acetyl alpha-MSH remained the major form. Five days after birth alpha-MSH and its diacetylated form constitute the major tissue and release form of the peptide, a situation very similar to that in adult mice. Acetylation of beta-endorphin appeared to occur earlier in development, N alpha-acetyl beta-endorphin (1-31) being the major form of endorphin already in the fetal pars intermedia. It is concluded that in the mouse acetylation of melanotropin and acetylation of beta-endorphin are not necessarily concomitant events. It could be established that the ability of the pars intermedia cells for cleaving N alpha-acetyl beta-endorphin (1-31) to yield C-terminally shortened forms of beta-endorphin develops after birth.     

Regulation of differential release of alpha-melanocyte-stimulating hormone forms from the pituitary of a teleost fish, Oreochromis mossambicus.

Using high-performance liquid chromatography (HPLC) in combination with radioimmunoassay, three forms of alpha-MSH (des-acetyl, mono-acetyl and di-acetyl alpha-MSH) were separated and identified in tilapia neurointermediate lobes and plasma, and in medium from lobes superfused in vitro. The presence of acetylated forms in lobe extracts indicated that the peptides are acetylated intracellularly. Di-acetyl alpha-MSH was, especially in comparison with monoacetyl alpha-MSH, relatively more abundant in lobe extracts than in plasma. This suggests that the three forms of alpha-MSH are not released according to their relative intracellular abundances. The possibility of regulation of this differential release by dopamine and TRH was investigated, using a microsuperfusion system. Dopamine was a potent inhibitor of alpha-MSH release, but did not modulate the relative abundance of the different forms of alpha-MSH released from the MSH cells. TRH was a potent stimulator of alpha-MSH release. It enhanced in vitro the release of di-acetyl alpha-MSH more than the release of mono-acetyl alpha-MSH. Thus tilapia may be able to modulate not only the quantitative but also the qualitative signal from the MSH cells. This might enhance the flexibility of the animals to respond to environmental challenges.     

Re-introduction and evaluation of an accurate, high capacity bioassay for melanocyte-stimulating hormone using the skin of Anolis carolinensis in vitro.

A simple and rapid assay for the quantitative determination of melanocyte-stimulating hormone (MSH) activity, using skin fragments of the lizard Anolis carolinensis in vitro, is described in detail. This assay, in which a quantal response (e.g. induction of a brownish-green colour) can be detected by visual observation, allows determination of MSH activity in up to 50 samples/day by one person, using the skin of one lizard. The mean dose found to induce this colour change was 0-15 ng synthetic alpha-MSH/ml (range 0-02-0-5 ng/ml). The assays shows high accuracy, reproducibility and specificity. Anterior and posterior lobe hormones as well as pituitary catecholamines do not interfere with the determination of pituitary MSH activitymthe method is compared with the assay using hypophysectomized frogs (Rana pipiens) in vivo. Determinations of MSH in pituitary extracts by both methods gave quantitatively similar results when determined with alpha-MSH as the reference substance. However, when beta-seryl MSH was used as a reference, the two assays gave different results for the MSH activity of the pituitary extractsmthis indicates that MSH from the rat pituitary gland has a biological activity similar to that of alpha-MSH rather than to that of beta-seryl MSH.     

Differences in the release of melanocyte-stimulating hormone in vitro by rat pituitary glands collected at various times during the oestrous cycle

The release of melanocyte-stimulating hormone (MSH) into the medium during incubation and the pituitary tissue content of MSH were measured separately using pituitary glands collected from rats at various stages of the oestrous cycle. The MSH was measured by a biological assay using a synthetic alpha-MSH as standard. The release of MSH was maximal during thepro-oestrous phase and MSH content of the pituitary gland was highest during dioestrus. The influences of the tripeptide Pro-Leu-Gly-NH2, which inhibits MSH secretion in vivo, and of progesterone on the release of MSH in vitro were studied with tissue collected at various phases of the oestrous cycle. Pro-Leu-Gly-NH2 was effective in inhibiting MSH release both at pro-oestrus and oestrus but not at dioestrus. Progesterone overcame this inhibition.     

Determination of tissue and plasma concentrations of PTHrP in fish: development and validation of a radioimmunoassay using a teleost 1-34 N-terminal peptide

A specific and sensitive radioimmunoassay (RIA) for the N-terminus of sea bream (Sparus auratus) and flounder (Platichthys flesus) parathyroid hormone-related protein (PTHrP) was developed. A (1-34) amino-terminal sequence of flounder PTHrP was synthesized commercially and used as the antigen to generate specific antiserum. The same sequence with an added tyrosine (1-35(Tyr)) was used for iodination. Human (1-34) parathyroid hormone (PTH), human (1-34) PTHrP, and rat (1-34) PTHrP did not cross-react with the antiserum or displace the teleost peptide. Measurement of PTHrP in fish plasma was only possible after denaturing by heat treatment due to endogenous plasma binding activity. The minimum detectable concentration of (1-34) PTHrP in the assay was 2.5 pg/tube. The level of immunoreactive (1-34) PTHrP in plasma was 5.2+/-0.44 ng/ml (mean+/-SEM, n=20) for flounder and 2.5+/-0.29 ng/ml (n=64) for sea bream. Dilution curves of denatured fish plasma were parallel to the assay standard curve, indicating that the activity in the samples was indistinguishable immunologically from (1-34) PTHrP. Immunoreactivity was present, in order of abundance, in extracts of pituitary, oesophagus, kidney, head kidney, gills, intestine, skin, muscle, and liver. The pituitary gland and oesophagus contained the most abundant levels of PTHrP, 37.7+/-6.1 ng/g wet tissue and 2.3+/-0.7 ng/g wet tissue, respectively. The results suggest that in fish PTHrP may act in a paracrine and/or autocrine manner but may also be a classical hormone with the pituitary gland as a potential major source of the protein.     

Alpha-MSH, the melanocortin-1 receptor and background adaptation in the Mozambique tilapia, Oreochromis mossambicus

The regulation of skin darkness in vertebrates is mediated by alpha-melanophore-stimulating-hormone (alphaMSH). For this action, alphaMSH binds to the melanocortin (MC)-1 receptor, a 7-transmembrane receptor located in melanophore cell membranes. The Mozambique tilapia, Oreochromis mossambicus, can change the hue of its body in response to a change in background, a process that may involve alphaMSH and the MC1R. Scale melanophores were isolated from tilapia that were acclimatised for 25 days to a black, control grey or white background and then tested for their sensitivity to des-, mono-, and di-acetylated alphaMSH. On all backgrounds, mono-acetylated alphaMSH was the dominant isoform present in pituitary homogenates. Mono-acetylated alphaMSH also had the highest potency to disperse melanosomes. Black background adapted fish showed the highest dispersing response to alphaMSH, independent of the isoform applied. We elucidated the nucleotide and amino acid sequence of the tilapia MC1R. We show that its expression in skin does not change when tilapia are acclimatised for 25 days to a black, grey or white background, while a clear change in hue is visible. This finding, combined with the absence of differential MC1R gene expression following background acclimation indicates that the increased sensitivity to alphaMSH is most likely a result of changes in the intracellular signalling system in melanophores of black background adapted fish, rather than up-regulation of the MC1R.     

A black background facilitates the response to stress in teleosts

This work examines the difference in responsiveness to stress which characterizes fish adapted to white and black backgrounds. Trout were maintained in black or white tanks for 2 weeks and then subjected to intermittent intense or moderate noise stress for periods between 1 h and 5 days, or to the stress of being injected daily with a large volume of liquid for 3 days. Plasma cortisol concentrations increased more readily and to a greater extent in fish from black tanks in response to moderate stress or brief intense stress. Dexamethasone suppressed the stress-induced rise of cortisol in white-adapted fish but was only partially effective in trout from black backgrounds. These differences in plasma cortisol between black- and white-adapted fish can be related to the different titres of plasma ACTH, apparently derived from the pars distalis. Removal of the neurointermediate lobe (NIL) from black-adapted eels markedly depressed the normal rise in plasma cortisol elicited by noise stress. It is suggested that products from the NIL may modulate the stress response of the hypothalamo-pituitary axis although other routes through which background colour could affect the pituitary responsiveness to stress are also considered. In several cases, stress also enhanced the secretion of MSH from the NIL.     

Molecular forms of alpha-melanocyte-stimulating hormone in pheochromocytoma tissue

The molecular forms of alpha-melanocyte-stimulating hormone (MSH) in pheochromocytoma tissues have been characterized using reversed-phase HPLC and radioimmunoassay. Six alpha-MSH-related peptides were detected. Three of the six peaks had elution times identical to those of synthetic desacetyl-alpha-MSH, alpha-MSH and diacetyl-alpha-MSH. The remaining three forms of the alpha-MSH-like immunoreactivity suggest a different processing of pro-opiomelanocortin in this tissue than in the intermediate lobe of the pituitary gland. Current results confirm that alpha-MSH are present in human pheochromocytoma and suggest that alpha-MSH has role in the pathophysiology of this tissue.     

Prolactin-releasing peptide, a possible modulator of prolactin in the euryhaline silver sea bream (Sparus sarba): A molecular study

PRL and PrRP cDNAs have been isolated from euryhaline silver sea bream (Sparus sarba). The PRL cDNA consists of 1360 bp encoding 212 amino acids whereas the PrRP cDNA contains 631 bp encoding preproPrRP with 122 amino acids. The mature PrRP sequence within the preprohormone is identical to the PrRPs isolated from other fish species. PRL mRNA was uniquely expressed in sea bream pituitary but PrRP mRNA was expressed in a variety of organs and tissues including the intestines, olfactory rosette and various brain regions such as hypothalamus and pituitary. Expression levels of PRL and PrRP mRNA have been examined in sea bream adapted to different salinities (0, 6, 12, 33 and 50 ppt). In the pituitary, both PRL and PrRP mRNA were significantly higher in fish adapted to low salinities (0 and 6 ppt) and the expression profiles of both hormones closely paralleled each other. However, expression of hypothalamic PrRP was significantly higher in fish adapted to iso-osmotic salinity (12 ppt) when pituitary PRL expression was low. The present study demonstrates, for the first time, a synchronized mRNA expression pattern between PRL and PrRP in fish pituitary but a disparity of mRNA expression levels between hypothalamic PrRP and pituitary PRL during salinity adaptation. These data suggest that PrRP may possibly act as a local modulator in pituitary rather than a hypothalamic factor for regulation of pituitary PRL expression in silver sea bream.     

Melanin concentrating hormone inhibits the release of alpha MSH from teleost pituitary glands

A radioimmunoassay was developed for salmonid melanin concentrating hormone (MCH) and used to measure immunoreactive (ir)MCH in the hypothalamus and pituitary of trout (Salmo gairdneri) and eels, (Anguilla anguilla) maintained under different regimes of background color. In trout, 95% of the total irMCH was located in the pituitary gland. The amount of MCH in both pituitary and hypothalamus was increased when white-adapted trout were transferred to a black background. In eels, a similar change of background led to an accumulation of MCH in the pituitary but not in the hypothalamus. The results suggest that MCH is released from the neurohypophysis in association with physiological color change. Neurointermediate lobes of trout and eels released both ir alpha MSH and irMCH when they were cultured in vitro. The release of alpha MSH was significantly enhanced when endogenous MCH was immunoabsorbed by MCH antiserum added to the culture medium. The results indicate that MCH can induce pallor in fish not only by its peripheral effect on the melanophores but also by an inhibitory action on the release of alpha MSH from the pituitary.     

Characterization of triacetyl-α-melanocyte-stimulating hormone in carp and goldfish

A triacetyl form of α-melanocyte-stimulating hormone (MSH) was found in carp (Cyprinus carpio) and goldfish (Carassius auratus), by selective detection of mass profile for cell secretory granules using direct tissue matrix-assisted laser desorption ionization with time-of-flight mass spectrometry (MALDI-TOF MS) analysis during the investigation of fish pituitaries. The structure of triacetyl-α-MSH in carp and goldfish was further analyzed using a collision-induced dissociation with electrospray ionization mass spectrometry, and determined to be N,O-diacetyl Ser as the N-terminal residue and O-acetyl Tyr at position 2. These modifications for α-MSH in carp and goldfish are structurally different from that of medaka hormone, in which [N,O-diacetyl Ser(1), O-acetyl Ser(3)]-α-MSH has been identified. The profiles of four α-MSH variants, des-, mono-, di- and tri-acetyl forms in goldfish and medaka pituitaries were also examined by direct tissue MALDI-TOF MS analysis, and the percentages as a total of α-MSH molecules were compared for fish reared in a white or black tank for 5 days. Among structural variants, diacetyl-α-MSH was the predominant form in goldfish and N-desacetyl-α-MSH in medaka, respectively. In both species, the relative level of the predominant form in the pituitary of white-adapted fish tended to be lower than that of black-adapted fish. In goldfish, no significant difference was observed in the relative content of triacetyl-α-MSH in both backgrounds, whereas the lowest content of triacetyl-α-MSH was found in black-adapted medaka. These preliminary data indicate that it is difficult to elucidate the relations between the physiological roles and acetylated pattern of α-MSH molecule, depending on species.