Determination of nonmammalian ghrelin.

Ghrelin is a peptide with a unique molecular modification by a fatty acid such as n-octanoic acid. The acyl modification is necessary for ghrelin to bind to its receptor (growth hormone secretagogue-receptor 1a, GHS-R1a) and to induce subsequent intracellular Ca(2+) signaling. Ghrelin is widely expressed in mammals as well as in nonmammalian vertebrates. In our laboratory, a method for efficient purification of ghrelin from a small amount of tissues has been established. Here, we introduce the identification process of ghrelin in nonmammalian vertebrates.

Neuroanatomical distribution of MCH in the brain and pituitary of submammalian vertebrates.

Melanin-concentrating hormone (MCH) is a cyclic neuropeptide that has been initially characterized from a salmon pituitary extract and subsequently identified in various species from all classes of vertebrates. The present review summarizes the current knowledge regarding the neuroanatomical distribution of MCH-immunoreactive neurons in submammalian vertebrates. In all species examined, MCH-immunoreactive perikarya are confined to the hypothalamus, with the exception of the cyclostome Lampetra fluvialis and the lungfish Protopterus annectens, in which additional populations of MCH-immunoreactive cell bodies occur in the telencephalon, and the frogs Rana ridibunda and Rana esculenta which exhibit MCH-positive perikarya in thalamic nuclei. In teleosts, in the frog R. ridibunda and in the L. fluvialis, MCH is present in the classical hypothalamic-neurohypophysial system indicating that the peptide may play the role of a neurohormone. In other groups, MCH-immunoreactive nerve fibers are widely distributed in various brain regions suggesting that, in these species, MCH in the central nervous system may act as a neurotransmitter or/and a neuromodulator rather than a neurohormone.

Soil ingestion may be an important route for the uptake of contaminants by some reptiles.

Some species of reptiles regularly ingest soil in the wild. Therefore, we evaluated the importance of soil ingestion as a route for the uptake of contaminants in lizards. We used sand as a substitute for soil during the present study. Different groups of leopard geckos (Eublepharis macularius) were provided with a control and five sodium selenite-spiked sand mixtures during a 28-d study. Twenty lizards were assigned to a control group and to each of five selenium-spiked sand mixtures that consisted of nominal selenium (Se) concentrations of 0.05, 0.46, 4.57, 11.41, and 22.83 mg Se/kg dry sand. Leopard geckos readily ingested the Se-spiked sand. We observed concentration-related effects in several endpoints. Overall growth in body mass was the most sensitive endpoint and was significantly (p < 0.05) lower in lizards that ingested the 4.57, 11.41, and 22.83 mg Se/kg sand mixtures compared to controls. Growth in snout-vent length, mean daily food ingestion, and food conversion efficiency were less sensitive and were significantly (p < 0.05) lower in lizards that ingested the 11.41 and 22.83 mg Se/kg sand mixtures compared to controls. Although our results are based on nominal amounts of Se ingested, leopard geckos appear to be as sensitive to sodium selenite as birds and mammals. The present study suggests that ingestion of soil could be an important potential route for the uptake of soil contaminants in some reptiles and should be evaluated in ecotoxicological studies and risk analyses of reptiles.

Physiological effects of a fish oil supplement on captive juvenile tuatara (Sphenodon punctatus).

Tuatara (Sphenodon, Order Sphenodontia) are rare New Zealand reptiles whose conservation involves captive breeding. Wild tuatara eat seabirds, which contain high levels of the long-chain n-3 polyunsaturated fatty acids (PUFAs) eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). These fatty acids are absent from the captive diet, and consequently, plasma fatty acid composition of wild and captive tuatara differs. This study investigated the effects of incorporating EPA and DHA into the diet of captive juvenile tuatara (Sphenodon punctatus) in an attempt to replicate the plasma fatty acid composition of wild tuatara. Tuatara receiving a fish oil supplement containing EPA and DHA showed overall changes in their plasma fatty acid composition. Phospholipid EPA and DHA increased markedly, reaching 10.0% and 5.9 mol%, respectively, by 18 mo (cf.

Protein and energy relationships in the diet of the American alligator (Alligator mississippiensis).

First-year alligators (Alligator mississippiensis) averaging 377-857 g body weight were fed diets containing various levels of protein, fat and carbohydrate. In experiment 1, nine diets arranged in a centrally rotatable composite design contained 0-36% extruded corn and 4-20% total fat. Response surface analysis predicted maximum responses in performance criteria at 6.3-18.8% corn and 15.8-27.4% fat. Corn inclusion at up to 27-36% of diet resulted in equal or improved performance compared to carbohydrate-free diets of equal fat content. Energy digestibility averaged 84.3%. Protein digestibility averaged 86.7%. Maximum responses in performance criteria were predicted at 42.5-48.7% digestible protein and 4367-4421 kcal/kg digestible energy. In two additional experiments, alligators were either fasted or fed for various numbers of days/week. Carbohydrate-supplementation of high protein diets led to equal or significantly improved performances. Performance was maximized by feeding the alligators 5-6 d/w. Regression of body weight changes against energy and protein intake yielded estimates of daily maintenance requirements of 5.7-8.4 kcal and 0.49-0.89 g protein/kg live body weight. Dietary fat and carbohydrate in the forms and amounts fed to young alligators were well-utilized. Optimal digestible energy:crude protein ratios (8.2-10.9:1 kcal/g protein) were similar to those of other aquatic ectotherms of equal size.

Changes in thyrotropin-releasing hormone levels in alligator stomach during fasting.

Stomach tissue of the American alligator, Alligator mississippiensis, contains substantial levels of thyrotropin-releasing hormone (TRH) which behaves identically to the synthetic hormone on radioimmunoassay (RIA) and high performance liquid chromatography (HPLC). Fasting induces a marked increase in gastric tissue levels of this hypophysiotropic hormone, but is without effect on hypothalamic content, suggesting a physiological role for TRH in gastric function of this vertebrate.

Activities of citrate synthase, NAD+-linked and NADP+-linked isocitrate dehydrogenases, glutamate dehydrogenase, aspartate aminotransferase and alanine aminotransferase in nervous tissues from vertebrates and invertebrates.

1. The activities of citrate synthase and NAD+-linked and NADP+-linked isocitrate dehydrogenases were measured in nervous tissue from different animals in an attempt to provide more information about the citric acid cycle in this tissue. In higher animals the activities of citrate synthase are greater than the sum of activities of the isocitrate dehydrogenases, whereas they are similar in nervous tissues from the lower animals. This suggests that in higher animals the isocitrate dehydrogenase reaction is far-removed from equilibrium. If it is assumed that isocitrate dehydrogenase activities provide an indication of the maximum flux through the citric acid cycle, the maximum glycolytic capacity in nervous tissue is considerably greater than that of the cycle. This suggest that glycolysis can provide energy in excess of the aerobic capacity of the tissue. 2. The activities of glutamate dehydrogenase are high in most nervous tissues and the activities of aspartate aminotransferase are high in all nervous tissue investigated. However, the activities of alanine aminotransferase are low in all tissues except the ganglia of the waterbug and cockroach. In these insect tissues, anaerobic glycolysis may result in the formation of alanine rather than lactate.