Erika M. Plisetskaya

The Effects of NPY and Insulin on Food Intake Regulation in Fish

Synopsis. Recent abundant studies report that in rodents starvation induces increased neuropeptide Y (NPY) mRNA expression and peptide secretion in the hypothalamus which reduces autonomic nervous activity and promotes food intake, and intracerebroventricular (ICV) injection of NPY has potent orexigenic effects. Conversely, the effect of insulin in the central nervous system is to inhibit food intake and NPY biosynthesis and secretion. In mammals body fatness is regulated and insulin acts as one intake inhibitory signal related to fatness. In salmon (Oncorhynchus sp.) we have demonstrated a rise in NPY-like mRNA expression and a coincident decrease in plasma insulin levels during 2 to 3 weeks of starvation. Additionally, experimentally manipulating body fatness with high and low fat diets has demonstrated that body fatness affects food intake in teleost fishes, raising the possibility that NPY and insulin act to regulate their food intake. Therefore, we hypothesized that as in rodents, ICV treatment with NPY would stimulate food intake while ICV insulin would reduce food intake. Preliminary results suggest that ICV NPY administration does stimulate food intake in channel catfish (Ictalurus punctatus), but central injection of insulin has no effect. Results of treatments with the sulfated octapeptide of cholecystokinin and the recombinant fragment of rat leptin 22–56 are also discussed.

Nutrient partitioning in rainbow trout at different feeding rates

Effects of feed deprivation (FR=0.0), underfeeding (FR=0.3% feed/day), moderately restricted feeding (FR=1.0) and adequate feeding (FR=2.0) on nutrient partitioning in 10-month-old rainbow trout (Oncorhynchus mykiss) were studied in a 6-week growth trial at a water temperature of 15°C. Each treatment was administered to triplicate groups of individually tagged rainbow trout initially weighing 0.32 kg. Average body weight gains and feed efficiencies were significantly (P<0.05) affected by the different feeding rates. Increased feeding rates had little influence on body composition except percent carcass fat and visceral protein. Percent fat in liver, red muscle and white muscle increased significantly with increasing feeding rates. Plasma protein, insulin, glucagon, and glucagonlike peptides also increased significantly, while growth hormone decreased, but not significantly, with increasing feeding rates.

Glucagon and Glucagon-like Peptides in Fishes

Glucagon and glucagon-like peptides (GLPs) are coencoded in the vertebrate proglucagon gene. Large differences exist between fishes and other vertebrates in gene structure, peptide expression, peptide chemistry, and function of the hormones produced. Here we review selected aspects of glucagon and glucagon-like peptides in vertebrates with special focus on the contributions made by analysis of piscine systems. Our topics range from the history of discovery to gene structure and expression, through primary structures and regulation of plasma concentrations to physiological effects and message transduction. In fishes, the pancreas synthesizes glucagon and GLP-1, while the intestine may contribute oxyntomodulin, glucagon, GLP-1, and GLP-2. The pancreatic gene is short and lacks the sequence for GLP-2. GLP-1, which is produced exclusively in its biologically active form, is a potent metabolic hormone involved in regulation of liver glycogenolysis and gluconeogenesis. The responsiveness of isolated hepatocytes to glucagon is limited to high concentrations, while physiological concentrations of GLP-1 effectively regulate hepatic metabolism. Plasma concentrations of GLP-1 are higher than those of glucagon, and liver is identified as the major site of removal of both hormones from fish plasma. Ultimately, GLP-1 and glucagon exert effects on glucose metabolism that directly and indirectly oppose several key actions of insulin. Both glucagon and GLP-1 show very weak insulinotropic activity, if any, when tested on fish pancreas. Intracellular message transduction for glucagon, especially at slightly supraphysiological concentrations, involves cAMP and protein kinase A, while pathways for GLP are largely unknown and may involve a multitude of messengers, including cAMP. In spite of fundamental differences in GLP-1 function between fishes and mammals, fish GLP-1 is as powerful an insulinotropin for mammalian B-cells as mammalian GLP-1 is a metabolic hormone if tested on piscine liver.

Insulin, insulin-like growth factor-I (IGF-I) and glucagon: the evolution of their receptors

Insulin and glucagon, two of the most studied pancreatic hormones bind to specific membrane receptors to exert their biological actions. Insulin-like growth factors IGF-I and IGF-II are structurally related to insulin, although they are expressed ubiquitously. The biological functions of the IGFs are mediated by different transmembrane receptors, which includes the insulin, IGF-I and IGF-II receptors. The interaction of insulin, insulin related peptides and glucagon with the corresponding receptors has been studied extensively in mammals and continues to be so. At the same time, research on ectothermic animals has made enormous progress in the recent years. This paper summarizes current knowledge on insulin, IGF-I and glucagon receptors, from a comparative point of view with special attention to non-mammalian vertebrates. The review covers adult and mostly typical target tissues, and with very few exceptions, developmental aspects are not considered. Binding characteristics, tissue distribution and structure of insulin and IGF-I receptors will be considered first, because both ligands and receptors are structurally related and have overlapping functions. These sections will be followed by similar distribution of information on glucagon receptors. Readers interested in either structure or functions of insulin, IGFs and glucagon in nonmammalian vertebrates are referred to other reviews (Mommsen TP, Plisetskaya EM. Insulin in fishes and agnathans: history, structure and metabolic regulation. Rev Aquat Sci 1991;4:225-259; Mommsen TP, Plisetskaya EM. Metabolic and endocrine functions of glucagon-like peptides: evolutionary and biochemical perspectives. Fish Physiol Biochem 1993;11:429-438; Duguay SJ, Mommsen TP. Molecular aspects of pancreatic peptides. In: Sherwood NM, Hew CL, editors, Fish Physiology. vol 13. 1994:225-271; Plisetskaya EM, Mommsen TP. Glucagon and glucagon-like peptides in fishes. Int Rev Citol 1996;168:187-257.).

Characterization of coho salmon (Oncorhynchus kisutch) islet somatostatins.

Three different somatostatins have been isolated from the pancreatic islet tissue of the coho salmon (Oncorhynchus kisutch) by gel filtration and HPLC. Two of these peptides contain 14 amino acids and the larger third peptide consists of 25 amino acids. The sequence of the salmon SST-25 is Ser-Val-Asp-Asn-Leu-Pro-Pro-Arg-Glu-Arg-Lys-Ala-Gly -Cys-Lys-Asn-Phe-Tyr-Trp-Lys-Gly-Phe-Thr-Ser-Cys. The sequence of the salmon SST-14-I is Ala-Gly-Cys-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys. The other small somatostatin (SST-14-II) which was not sequenced has an amino acid composition identical to the C-terminal 14 amino acids of the SST-25 and it is probably derived from this larger form. Evidence for low levels of a somatostatin containing 28 amino acids is also presented. This SST-28 appears to be an N-terminal extended precursor of SST-25 or a peptide derived via alternative processing of a common preprosomatostatin. Injected into juvenile salmon, SST-25 caused a decline in circulating levels of plasma insulin, depletion of liver glycogen, and activation of lipolytic pathways. Juvenile salmon treated with anti-SST-25 serum revealed elevated levels of plasma insulin as well as an increase of the glycogen content of the liver.

Regulation of nutrient intake and energy balance in salmon

There is a correlation between growth, adiposity and early maturity in salmonids. One strategy to avoid early maturity may be to favor muscle growth over fat accumulation. The present article discusses the regulation of food intake and fat storage under conditions of either relative over-abundance or insufficiency of energy intake in salmon. In a series of experiments with chinook salmon of equal size, food intake of fat fish was significantly less than in their lean counterparts. Furthermore, when fat and lean fish were pooled and given access to the same diet, whole body fat levels converged, suggesting a robust mechanism for regulating body fat stores. Growth of the initially fat fish was impaired relative to the initially lean fish. In separate experiments, when chinook and coho salmon were deprived of food, hypothalamic expression of neuropeptide Y (NPY)-like mRNA increased significantly compared to fed controls. Increased hypothalamic NPY mRNA and NPY secretion have been shown to be central elements of energy balance regulation in mammals. The increase in hypothalamic NPY-like gene expression in a teleost may reflect the primitive evolution a central mechanism to regulate energy intake and expenditure that has been elucidated in mammals.

Effects of somatostatin-25 and urotensin II on lipid and carbohydrate metabolism of coho salmon, Oncorhynchus kisutch

Salmon (Oncorhynchus kisutch) somatostatin (sSS; 4 or 8 ng/g body wt) or synthetic Gillichthys urotensin II (UII; 2 or 4 ng/g body wt) were injected intraperitoneally into juvenile freshwater coho salmon. Both sSS and UII caused a dose-dependent increase in plasma free fatty acids (FFA) which diminished with time. sSS induced an initial (1 hr) transient hyperglycemia. By contrast, UII tended to induce hypoglycemia, this effect being significant 5 hr after injection of the higher dose. Both sSS and UII depressed plasma insulin titers 1 hr after injection. By 3 hr, the sSS-associated insulin depression was no longer observed. UII treatment induced a hyperinsulinemia which was present 3 and 5 hr after peptide administration. Although no decreases in liver total lipid concentration or in mesenteric fat total tissue mass were observed, lipolytic enzyme activity within each depot was significantly enhanced by both peptides. Neither sSS nor UII altered 3H2O incorporation into fatty acids or neutral lipids. However, enhanced lipogenesis, particularly by UII, was indicated by increased NADPH production resulting from glucose-6-phosphate dehydrogenase activity. Both sSS and UII enhanced glucose mobilization, as indicated by decreased liver glycogen content and increased liver glucose-6-phosphatase activity. UII, but not sSS, stimulated glycogen synthetase activity. These results suggest that both sSS and UII stimulate hyperlipidemia by enhancing depot lipase activity and that although both factors are potentially gluconeogenetic, sSS seems to be glycogenolytic and hyperglycemic, whereas UII may channel glucose to FFA synthesis.

Thyroid hormones in cyclostomes and fish and their role in regulation of intermediary metabolism

1. Experimental data obtained in cyclostomes and fish concerning the plasma levels of thyroxine and tri-iodothyronine as well as their influence on intermediary metabolism of lipids, carbohydrates, and proteins are reviewed. 2. The information dealing with the physiological role of thyroid hormones in regulation of metabolic processes seems to be scarce in cyclostomes and controversial in fishes. 3. Nevertheless, the data covered in the review support the generalization that thyroid hormones, probably along with some other hormones, exert a regulatory action on the metabolic processes already on the lower stage of the evolution of poikilothermic vertebrates.

Effects of injected and dietary arginine on plasma insulin levels and growth of Pacific salmon and rainbow trout

1. 1. Amino acids (arginine, alanine and lysine) injected intraperitoneally 6.6 μmol/g body weight elevate, while histidine decreases plasma circulating levels of insulin in coho salmon (Oncorhynchus kisutch). 2. 2. Insulinotropic action of arginine (0.03–6.6 μmol/g body weight) can be observed at 0.5–5.0 hr after injection. The effect of arginine is dependent on the season being more prolonged in winter when water temperature is low. 3. 3. Insulinotropic action of arginine seems to be direct rather than through stimulation of glucagon or glucagon-like peptide secretion. 4. 4. Feeding coho salmon, Chinook salmon (O. tshawytscha) and rainbow trout (O. mykiss) diets supplemented with arginine caused elevation of plasma insulin levels. Arginine supplemented diets (20–30 g/kg dry diet over the content of regular diet) had a short-lasting stimulatory effect on fish growth, an effect which could be detected only in fingerlings.

Does oral 3,5,3 ′-triiodo-l-thyronine affect dietary glucose utilization and plasma insulin levels in rainbow trout (Salmo gairdneri)?

A factorial experiment was conducted to determine the effect and interaction of dietary carbohydrate level and triiodo-L-thyronine (T3) supplementation on the growth, physiological response and plasma insulin and cortisol levels of rainbow trout. The oral administration of T3 significantly increased the growth, protein efficiency ratio and feed efficiency of trout, indicating an increased protein and perhaps energy utilization in these fish. However, T, administration did not significantly increase the utilization of dietary glucose as an energy source by the trout. Similarly, the administration of T3 did not significantly affect plasma insulin levels in either the fed or the fasted trout. Plasma insulin levels were significantly higher in fed trout reared on the non-T3 supplemented high carbohydrate diet in comparison to trout reared on the low carbohydrate diets. This indicates that increased dietary carbohydrate stimulates increased insulin secretion in the trout. Therefore, although rainbow trout are not insulin-deficient, they can still be considered a diabetic-like animal due to their poor glucose tolerance. Plasma cortisol levels were not affected by diet composition and altered plasma glucose levels.