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The isolation and chemical characterization of a novel vasoactive intestinal peptide-related peptide from a teleost fish, the cod, Gadus morhua.

An octacosapeptide that shows sequence homology to porcine vasoactive intestinal peptide (VIP) has been isolated from a teleost fish, the cod, Gadus morhua. The full primary sequence is His-Ser-Asp-Ala-Val-Phe-Thr-Asp-Asn-Tyr-Ser-Arg-Phe-Arg-Lys-Gln-Met-Ala-Ala- Lys-Lys-Tyr-Leu-Asn-Ser-Val-Leu-Ala. This peptide contains four, or five substitutions, compared with dogfish and porcine VIP, respectively. The residues in positions 13, 26 and 28 are different in all three species. These substitutions seem to have little effect on bioactivity, since cod VIP was virtually equipotent with porcine VIP in stimulating amylase release from guinea-pig pancreatic acini. During the isolation procedure an N-terminally modified form of VIP (Des-His, or 2-28 cod VIP) was also isolated. The available data suggest the sequence of VIP is well conserved in vertebrates which is consistent with an important biological role.

Amino Acid Sequence↗

The hemoglobins of the cold-adapted Antarctic teleost Cygnodraco mawsoni.

The blood of the teleost Cygnodraco mawsoni, of the endemic Antarctic family Bathydraconidae, contains a major hemoglobin (Hb 1), accompanied by a minor component (Hb 2, about 5% of total). The two hemoglobins have identical alpha chains and differ by the beta chain. The complete amino acid sequence of the three chains has been elucidated, thus establishing the primary structure of both hemoglobins. The sequences show a 53-65% identity with non-Antarctic poikilotherm fish species; on the other hand, a very high degree of similarity (83-88%) has been found between Hb 1 and the major component of another Antarctic species of a different family. The hemoglobin functional properties relative to oxygen binding have been investigated in intact erythrocytes, 'stripped' hemolysate and purified components of C. mawsoni. The hemoglobins display the Bohr and Root effects, indicating fine regulation of oxygen binding by pH and by the physiological effectors organic phosphates.

Adaptation, Physiological↗

Glucose-6-phosphate dehydrogenase from the blood cells of two antarctic teleosts: correlation with cold adaptation.

Glucose-6-phosphate dehydrogenase (G6PD) has been purified from the blood of two Antarctic teleost species, i.e., from the erythrocytes of Dissostichus mawsoni (family Nototheniidae), and from the plasma and cells of haemoglobinless Chionodraco hamatus (family Channichthyidae). The specific activities in haemolysates of Antarctic blood cells appear higher than that of a lysate of human erythrocytes. The two Antarctic enzymes have an apparent subunit molecular mass slightly higher than that of human G6PD; the electrophoretic behaviour on cellulose acetate is similar. Both Antarctic enzymes are irreversibly heat inactivated through a biphasic process. Km for glucose-6-phosphate (G6P) does not vary significantly with temperature, whereas Km for NADP increases at increasing temperature, kcat increases with temperature, with a break point at 35 degrees C (in human G6PD, the break point is at 15 degrees C). Thermodynamic and kinetic characterisation indicate that the catalytic performance of the enzyme of cold-adapted fish, at temperatures typical of their habitat, is more efficient than that displayed by G6PD from a temperature organism.

Adaptation, Physiological↗

An investigation on pro-opiomelanocortin and processed peptides from the teleost fish Prochilodus platensis.

Acid extracts of carefully dissected proadenohypophysis (PA) and metaadenohypophysis (MA) of the teleost Prochilodus platensis were subjected to chromatography in Sephadex G-50 after which several pro-opiomelanocortin (POMC) peptides were detected by means of three heterologous RIA systems: alpha-MSH, ACTH and beta-endorphin. Parallelism among extracts displacement curves ranged from 26% to 95% of those of the standard curves for the different systems employed. In PA chromatograms, peaks of ACTH immunoreactivity (IR) were detected at the positions of 30 kilodalton (K), 20K, 9K, a large 4.5K peak and 2K. Only one peak of beta-endorphin IR was detected at 30K. In MA chromatograms, ACTH IR detected similar peaks as in PA runs, but 4.5K peak was much smaller, whereas a large 2K peak roughly coincided with all alpha-MSH detected in the chromatograms. beta-Endorphin IR was detected mainly as a large peak coinciding with synthetic beta-endorphin in MA runs. Bioactivity was detected in both PA and MA 4.5K ACTH peaks, whereas little activity could be demonstrated associated with the 30K, 20K and 9K ACTH IR peaks. Prochilodus PAs and MAs were incubated with tritiated aminoacids and the extracts immunoprecipitated with ACTH, beta-endorphin and N-terminal POMC (N-POMC) antisera. The dissociated complexes were run in SDS polyacrylamide slab gel electrophoresis. The tritiated bands detected confirmed the results obtained with Sephadex chromatography. N-POMC immunoprecipitated peptides were located at 28K, 18K and 9K positions. The first two probably accounted for POMC and the N-POMC/ACTH intermediate respectively; the third corresponded to the mammalian 1-76N-POMC.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenocorticotropic Hormone↗

Pancreatic peptides regulate Cl- secretion in the marine teleost gill.

Hormonal regulation of the ionoregulatory function of the branchial epithelium in marine teleosts has been investigated using an isolated gill preparation of the seawater-adapted flounder Platichthys flesus. Agents which are assumed to elevate the effective intracellular concentration of cyclic AMP caused a stimulation of Cl- secretion as determined by measurement of the transepithelial electrical potential difference in the isolated gill. The pancreatic endocrine glucagon may act as an endogenous regulator of ion excretion in the gill as this peptide stimulated the transepithelial potential. Another islet peptide, somatostatin, inhibited branchial electrogenic ion transport. Attempts to investigate the mode of action of these peptides by measuring cyclic AMP levels in isolated gills were unsuccessful, since it appears that the pool of cyclic nucleotide which specifies the epithelial Cl- transport rate is small compared to total cyclic AMP content. A second approach investigating the ability of somatostatin to inhibit the transepithelial potential in gills stimulated by different secretagogues suggested dual modes of action for this peptide.

Animals↗

Partial purification and characterization of a novel growth hormone-releasing factor (GRF) from teleost brain related to the rat hypothalamic peptide.

A growth hormone-releasing factor (GRF)-like molecule has been partially purified and characterized from acid extracts of codfish (Gadhus morhua) brain using immunoaffinity and gel chromatography, followed by HPLC. This material has a mol.wt. which is similar to known mammalian forms of GRF but is immunologically and/or chromatographically distinct from previously described GRF peptides. However, it is related to rat(r) GRF(1-43) since it causes marked displacement in the rGRF RIA. Codfish GRF is a highly specific and potent hypophysiotropic factor as shown by its ability to stimulate the release of GH, but no other hormone, from rat anterior pituitary cells in vitro. These findings suggest that, phylogenetically, GRF is an ancient molecule with its biologic activity and certain immunoreactive domain(s) conserved, at least, from teleost to mammal.

Animals↗

Isolation and primary structure of gastrin-releasing peptide from a teleost fish, the trout (Oncorhynchus mykiss).

Immunohistochemical studies have established that fish gastrointestinal tissues contain peptides with gastrin-releasing peptide (GRP)/bombesin-like immunoreactivity, but the molecular nature of this material is unclear. In this study, the most abundant peptide that was immunoreactive towards an antiserum raised against pig GRP was isolated in pure form from an extract of the stomach of the rainbow trout (Oncorhynchus mykiss). The primary structure of the peptide was established as: Ser-Glu-Asn-Thr-Gly-Ala-Ile-Gly-Lys-Val10- Phe-Pro-Arg-Gly-Asn-His-Trp-Ala-Val-Gly20-His-Leu-Met-NH2. Although this amino acid sequence is shorter than those of mammalian GRPs by four residues, the COOH-terminal dodecapeptide is identical to the corresponding region in pig GRP. The data indicate, therefore, that the predominant molecular form of GRP in the stomach of a teleost fish is structurally more similar to mammalian GRP than to the amphibian skin peptide, bombesin.

Amino Acid Sequence↗

Effect of water pH and salinity on the survival of eggs and larvae of the euryhaline teleost, Gasterosteus aculeatus L.

The effect of water acidification and salinity on the survival of the eggs and larvae of the euryhaline teleost, Gasterosteus aculeatus, was studied using tap water that was low in calcium and high in total aluminium as the basic medium. The survival of eggs from fertilisation to hatching increased with an increase in pH from 4.5 to 6.5. Survival was higher at a given pH at higher salinities (equivalent to 5% and 10% seawater) than in tap water. The salinity experienced by the female prior to spawning also had an effect on egg survival. Survival for 7 days after hatching was high except at the lowest pH levels. The salinity experienced by the female had no significant effect on larval survival.

Journal Article↗

Effects of long-term exposure to cythion on the reproduction of the teleost fish, Channa punctatus (Bloch).

The chronic toxic effects of an organophosphorus pesticide on some aspects of the physiology of reproduction in a teleost fish, Channa punctatus, are described in this investigation. Fish exposed to 2.0 mg litre(-1) of commercial cythion continuously for 6 months, from January (resting phase) to June (spawning phase), exhibited a significant inhibition of gonadal development and gonadosomatic index. These changes were correlated with less active, and a reduced number of, pituitary gonadotrophs, and biochemical changes in the gonads and liver. The immature oocytes of the experimental fish exhibited cytoplasmic proteinaceous 'inclusion bodies', which ultimately led to their degeneration. The results suggest that cythion may act directly on the gonads and/or through the pituitary-gonadal axis, in this species.

Journal Article↗

Effect of vegetable oil factory effluent on the gonads of a freshwater teleost, Heteropneustes fossilis (Bl.) --a biochemical study.

The effects of three sublethal concentrations (5,10,15% v/v) of vegetable oil factory effluent on biochemical changes in the gonads (ovaries/testes) of a freshwater teleost were studied. The results reveal a significant decline in the total lipid, cholesterol, phospholipid, and glycerine levels, but an increase in the free fatty acid levels, both in the ovaries and the testes of H. fossilis, following 150 days (mid-January through mid-June) of exposure to 10% and 15% concentrations of the effluent. This decrease/increase in the biochemical parameters was more or less dependent on the concentration of the effluent. The functional significance of any decrease/increase in the biochemical parameter(s) has been discussed.

Journal Article↗

Effects of chromium(VI) on some ion-dependent ATPases in gills, kidney and intestine of a coastal teleost Periophthalmus dipes.

The coastal teleost species, Periophthalmus dipes, commonly known as the mudskipper, was exposed to three sublethal concentrations (5, 10 and 15 mg/l) of potassium chromate for three exposure durations (2, 4 and 6 days). The study compares the dose- and duration-dependent effects of Cr(VI), as potassium chromate, on the ATPase systems in various organs of this fish species. In this study, effects of Cr(VI) stress on total ATPase, (Na+,K+)-ATPase, (Ca+2)-ATPase, (Mg+2)-ATPase, (Ca+2, HCO3-)-ATPase and (Mg+2,HCO3-)-ATPase in gills, kidney and intestine were estimated. A general dose- and duration-dependent inhibitory trend was observed. However, it is evident that exposure duration is more important then dose in the inhibition of the activity of the enzymes. At some concentrations, initial stimulation of the activity of some enzymes were also noticed. However, maximum inhibition was observed in higher Cr(VI) concentrations exposed for the longest time. It is possible that this inhibition of the ATPases by Cr(VI) blocked the active transport system of the gill epithelial as well as chloride cells, glomerular and epithelial cells of the tubules and thus altered the osmoregulatory mechanism of the fish. It appears that this heavy metal ion alters the membrane permeability of the intestinal epithelial cells and other layer of cells by altering the activity of ATPases, resulting in a breakdown of the active transport mechanism needed for the absorption of nutrients, ions and metabolites.

Absorption↗

The effects of lowered temperature on spontaneous eye movements in a teleost fish.

A new opto-electronic method has been used to measure spontaneous eye movements in a lightly restrained unanaesthetized marine teleost fish (Parore). The normal scanning pattern of eye movement is similar to that previously described in goldfish. The effects of cooling on eye movements were investigated by 2 degrees C step changes down from ambient temperature (13-14 degrees C). Lowered temperature altered the scanning pattern, decreased saccade velocity, increased mean saccade amplitude and impaired the ability of the fish to hold the eye stationary between saccades. All eye movements stopped at temperatures around 6 degrees C, but could be restored by subsequent warming.

Animals↗

Functional characterization of hemoglobins from South Brazilian freshwater teleosts--I. Multiple hemoglobins from the gut/gill breather, Callichthys callichthys.

The five main hemoglobins of the South Brazilian bimodal breathing teleost, Callichthys callichthys were separated, and their oxygenation properties and those of the unfractionated hemolysate were measured at 10, 20 and 30 degrees C. The cathodal Hb component had a higher O2 affinity than the other components and a lower Bohr effect (phi = delta log P 50/delta pH), which is reversed at low and high pH values (6.8 greater than pH greater than 7.8). The other, anodal hemoglobins had normal Bohr effects and similar functional properties. The erythrocytic cofactor ATP had no significant effects on the O2 affinities of the hemolysate and the isolated anodal components at physiological pH conditions, but decreased the affinity of the cathodal Hb over the entire pH range tested (6.5-8.2). All hemoglobins showed high thermal dependence of O2 affinity (delta H values between -62 and -74 kJ mol-1 at pH 8.0), which decreased with falling pH, in accordance with an inverse relation between the Bohr effect and temperature. The possible adaptive significance of the oxygenation patterns of the hemoglobins and their temperature dependences are discussed comparatively with special reference to the closely-related bimodal breather Hoplosternum littorale, and to breathing habit (gill breathing in winter when the fish is a benthic feeder and "gut" air breathing in the warm reproductive season when they nest at the surface).

Animals↗

Aldosterone response to renin, angiotensin, ACTH, hemorrhage and sodium depletion in a freshwater teleost, Catostomus macrocheilus.

1. Plasma aldosterone activity (PAA) was measured in individual large-scale suckers (Catostomus macrocheilus) by radioimmunoassay. 2. PAA in control groups was 12.0 +/- 6.0 (mean +/- SEM) pg/ml; one control group showed a sixfold elevation of PAA, possibly reflecting a seasonal effect. 3. Injections of sucker renin preparation, synthetic angiotensin II and synthetic adrenocorticotrophin had no effect upon PAA. PAA was also unaffected by sodium depletion, sodium loading and moderate hemorrhage. 4. Prolonged restraint stress coupled with severe hemorrhage significantly increased PAA to 47.1 +/- 5.1 pg/ml. 5. This study indicates that aldosterone secretion in teleost fish has either rudimentary or yet to be discovered controls and functions.

Adrenocorticotropic Hormone↗

The development of a homologous teleost insulin radioimmunoassay and its use in the study of adrenaline on insulin secretion from isolated pancreatic islet tissue of the rainbow trout, Salmo gairdinerii (R.).

A homologous teleost insulin radioimmunoassay (RIA) employing bonito insulin RIA components is described. The RIA sensitivity and specificity was sufficient to measure endogenous immunoreactive insulin (IRI) levels in trout serum, pancreatic islet tissue extract and in vitro culture medium. Adrenaline at 10(-6)M evoked a marked stimulation of basal insulin release from isolated islets; whereas adrenaline at 10(-10)M evoked an inhibition of basal insulin release.

Animals↗

Occurrence of magnetic material in teleosts.

Fishes representing the main groups of teleosts have been investigated for magnetic material by susceptibility measurements. All the investigated species contain magnetic material. Bone samples from the skull and the vertebral column contain magnetic particles which yield a saturation magnetization in the range 10(-4) emu g-1 to 10(-3) emu g-1 in a sample. The localization of the magnetization is diffuse within the tissues connected to the bone. There are no significant differences between the amounts of magnetic material that are found in migrating or more stationary species.

Animals↗

Spectral sensitivity of melanophores of a freshwater teleost, Zacco temmincki.

1. The melanophores of a freshwater teleost, Zacco temmincki, responded to changes in illumination: in darkness the melanophores induced a melanosome aggregation and when subjected to light they caused a melanosome dispersion. 2. Using monochromatic light, the spectral sensitivity of the melanophores was examined. 3. The melanophores showed a different sensitivity to light between 400 and 600 nm with a maximum at about 525 nm. 4. The action spectrum closely resembled a porphyropsin absorbance curve, suggesting a porphyropsin or similar photopigment is active in the melanophore light response of Zacco temmincki.

Animals↗

Influence of blood sample collection on the haematocrit value of two teleosts: rainbow trout (Oncorhynchus mykiss) and European sea bass (Dicentrarchus labrax L.).

1. The effects of the volume of blood extracted, number of extractions, and weight upon the haematocrit value of a freshwater teleost (Oncorhynchus mykiss) and a marine one (Dicentrarchus labrax) have been studied. 2. For trout with the same weight, the increase in the volume of blood extracted results in a significant increase in the haematocrit value. 3. For the same volume of blood extracted--in trout of the same weight--the second extraction results in a significant increase in the haematocrit value. 4. In sea bass, haematocrit variations in relation to the aforementioned parameters are not produced. 5. For both species there is a positive and significant correlation between weight (W) and haematocrit value (Ht), which coincides with the Ht = aWb model, in the range of weights used in this study.

Animals↗