Biosynthesis of rainbow trout (Salmo gairdnerii) hemoglobins in vitro

Cell free systems were established to analyze the biosynthesis of trout hemoglobins I, II and III. HbI, II and III were synthesized in trout erythroid cell lysates as well as in a mRNA dependent rabbit reticulocyte lysate supplemented with trout erythroid cell polyA-RNA. The newly synthesized hemoglobins apparently contained the N alpha-acetyl modification at their alpha-chain amino terminals since they co-migrated with carrier trout hemoglobins that are known to contain the modification. This observation suggests that the acetylation is determined by the information encoded in the mRNA.     

Organization of the histone genes in the rainbow trout (Salmo gairdnerii)

Summary Twelve clones containing histone genes were isolated from a genomic trout library constructed in the vector Charon 4A. Each of the clones was found to contain a conserved 10.2-kb Eco RI fragment that contained one copy of each of the histones in the order H4-H2B-H1-H2A-H3, all of which are transcribed from the same strand. Genomic Southern blots indicate that these clusters are representative of the vast majority of the histone genes in the trout. Tandemly linked clusters were not found. Approximately 145 copies of this cluster are present in a trout sperm cell. Sequence analysis has shown the genes to be without introns and to show strong selection for codons ending in C or G. Consensus signals similar to those found in other histone genes are present in the flanking regions.     

Influence of dietary composition on gluconeogenesis from L-(U-14C)glutamate in rainbow trout (Salmo gairdneri)

The effect of dietary composition (high-protein, high-carbohydrate and high-fat diets) and starvation on in totum gluconeogenesis from L-(U-14C)glutamate was studied in the rainbow trout. High-fat and high-carbohydrate diets produced a significant hyperglycaemia. Lower blood glucose values were obtained in trout fed on a high-protein diet. Liver glycogen levels were significantly lower in trout fed on carbohydrate-free diets (high-protein and high-fat diets) and in starved fish. Gluconeogenesis from L-(U-14C)glutamate was markedly reduced in fish given the high-carbohydrate diet and significantly enhanced in starved fish. Radioactive liver glycogen was higher in starved fish, although the amount of radioactivity incorporated into glycogen was very low.     

Nutritional regulation of glycolysis in rainbow trout (Salmo gairdneri R.)

1. Some physico-chemical constants and the nutritional regulation of pyruvate kinase (PK), phosphofructokinase (PFK) and hexokinase (HK) from rainbow trout liver was investigated. 2. The maximum activity pH for the three enzymes appears to be in a physiological range. 3. The PK-enzyme shows sigmoid kinetic with respect to PEP with a Hill-coefficient of 3.1; the other two enzymes show michaelian kinetic for their substrates. 4. The nutritional treatments show that HK-enzyme increases its level with high carbohydrate diet and decreases with high protein diet and starvation. 5. PFK-enzyme decreases with high protein diet and starvation. 6. PK-enzyme only shows a decrease in level with starvation conditions.     

Changes in plasma lipoproteins and tissue lipoprotein lipase and salt-resistant lipase activities during spawning in the rainbow trout (Salmo gairdnerii R.)

1. Lipoprotein lipase and salt-resistant lipase activities increased in the ovaries but decreased in the adipose tissue of female trout in the months leading up to spawning. 2. The activity of the plasma cholesterol esterifying enzyme increased significantly immediately prior to spawning. 3. Plasma lipoprotein concentrations decreased during the approach to spawning. 4. These studies suggest that the developing ovaries in the trout receive their nutrients by lipolysis of plasma lipoproteins as well as by vitellogenin uptake; differentiation of the roles of the lipid stores in different tissues is proposed.     

Nucleotide sequence of 5S ribosomal RNA from rainbow trout (Salmo gairdnerii) liver.

Staring from low molecular weight RNA obtained from rainbow trout (Salmo gairdnerii) liver, 5S ribosomal RNA (rRNA) was highly purified by successive chromatography on columns of DEAE-Sephadex A50 and Sephadex G100. Products of complete and partial digestions on this RNA with pancreatic ribonuclease (RNase A) [EC 3.1.4.22] and RNase T [EC 3.1.4.8] were isolated and sequenced by conventional and high-performance liquid chromatography (HPLC) procedures. The nucleotide sequence of this RNA thus established was compared with those of five other vertebrae 5S rRNAs, and the rates of base substitution per site per year were found to be nearly constant in these RNAs. The analyses of the partial digests of the trout 5S rRNA revealed several sites susceptible to RNase attack, which could be accounted for by the secondary structure model for eukaryotic 5S rRNAs proposed by Nishikawa and Takemura (1).     

Effects of partial hepatectomy on various blood and hepatic parameters in the rainbow trout (Salmo gairdnerii, Rich.)

After 36% of hepatic mass removal , rainbow trout recovered its initial liver weight in 20-30 days, i.e., with a regeneration rate clearly lower than in mammals. During early regeneration process hematocrit index and hemoglobin content were slightly decreased, but both parameters rapidly reached their normal values. The evolution of both glycaemia and hepatic glycogen content supported the idea of the existence of a late regeneration wave, which, in this case, could begin at about the 20th post-operative day.     

Viability control and oxygen consumption of isolated hepatocytes from thermally acclimated rainbow trout (Salmo gairdnerii)

Hepatocytes were isolated by collagenase perfusion of the liver from rainbow trout acclimated to 10 and 20 degrees C. The suitability of the stimulation of cellular respiration by succinate as criterion of viability was examined and discussed. Endogenous respiration rates of the hepatocytes were a function of cell size to the power of 0.8. Specific oxygen consumption of the hepatocytes and respiratory control ratios of the mitochondria in situ were independent of acclimation temperature.     

Biliary excretion of [14C]taurocholate by rainbow trout (Salmo gairdneri) is stimulated at warmer acclimation temperature

Sexually immature Shasta strain rainbow trout (Salmo gairdneri) of either sex were acclimated at 10, 14 or 18 degrees C for at least 4 weeks and plasma pharmacokinetics and biliary excretion of i.p. injected [14C]taurocholate (TC) examined in spinally transected or free-swimming fish, respectively. Plasma elimination half-lives but not absorption rate constants for [14C]TC (10 mumol/kg) were about two-fold reduced in 18 as compared to 10 or 14 degrees C acclimated fish. Distribution of [14C]TC to tissues other than plasma, liver, bile and small intestine was not different in 10, 14 or 18 degrees C acclimated free-swimming fish at 1 or 4 hr post-injection. Biliary excretion of [14C]TC (7.5-10 mumol/kg) at 1 hr post-injection was significantly higher in 14 and 18 as compared to 10 degrees C acclimated fish.     

Phosphofructokinase during haemopoiesis of the rainbow trout (Salmo gairdneri R.): its isoenzymatic forms

Phosphofructokinase (EC 2.7.1.11) from trout haemopoietic cells and erythrocytes exhibits biphasic behaviour, with respect to fructose 6-phosphate and MgATP in extracts filtered through Sephadex G-25. Two different values of Hill coefficient and S0.5 have been found for each cellular population. Two forms of the enzyme with high and low affinity for the substrates, were obtained after affinity chromatography in each cellular population. The kinetic behaviour of these forms is different and may be due to a distinct composition and/or proportion and contribution of phosphofructokinase isoenzymes in both types of cells.     

Hormonal effects on gluconeogenesis from (U-14C)glutamate in rainbow trout (Salmo gairdneri)

Blood glucose was significantly decreased by insulin (4 I.U./kg). Glucagon (1 mg/kg) and Cortisol (5 mg/kg) administration produced a significant hyperglycaemia. Insulin administration did not modify liver glycogen levels. Glucagon showed a marked liver glycogen mobilization. Cortisol stimulated liver glycogen deposition. Insulin and Glucagon showed a significant inverse effect on gluconeogenesis from (U-14C)glutamate, decreasing and increasing 14C-glucose formation respectively. Hormonal treatments did not influence the very low levels of incorporation of (U-14C)glutamate into liver and muscle glycogen.     

Na transport and enzyme activities in the intestine of the freshwater and sea-water adapted trout (Salmo gairdnerii R.)

Unidirectional fluxes of Na+ obtained in perfused preparation and mucosal enzyme equipment (alkaline phosphatase, ouabain-sensitive Na+, K+-ATPase) have been determined in the middle and posterior intestine of freshwater (FW) and sea-water (SW) adapted trout. In FW, influxes and outfluxes were higher in the middle than in the posterior intestine, although net fluxes were similar. SW adaptation induced an increase of influxes and net fluxes mainly in the posterior intestine. SW adaptation decreased the alkaline phosphatase activity only in the posterior intestine. Na+,K+-ATPase activity was always higher in the middle than in the posterior intestine in FW and SW and increased in both parts by SW adaptation. Thus, it seems that SW adaptation of rainbow trout modifies Na intestinal absorption principally in its posterior part and in relation with the Na+, K+-ATPase activity.     

In vivo protein synthesis in different tissues and the whole body of rainbow trout (Salmo gairdnerii R.). Influence of environmental temperature

The fractional protein synthesis rate (FSR) of tissue (liver, digestive tract, muscle and whole fish) proteins was measured in rainbow trout acclimated to 9 and 18 degrees C after a pulse injection of [U-14C] L-leucine. In each of the tissues two FSRs were calculated based on a different estimate of the specific radioactivity of leucine in the precursor compartment for protein synthesis. Whole fish protein synthesis (WFPS) was estimated to be 7 and 7.6 g protein per kg body weight and per day respectively at 10 and 18 degrees C. Muscle and digestive tract contributed the most (more than 30%) to WFPS. The rate of protein turnover in whole fish was very low, as in the muscle, when compared to liver and digestive tract.