Spawning induces a shift in energy metabolism from glucose to lipid in rainbow trout white muscle

Enzymatic changes that occur in the white somatic muscle of rainbow trout (Oncorhynchus mykiss) in response to spawning were investigated, and the evenness of their distribution across the ventral-dorsal plane of this muscle was assessed. Four enzymes that are involved in energy metabolism were measured (phosphofructokinase: glycolytic capacity, 3-hydroxyacyl-CoA dehydrogenase: -oxidation, citrate synthase: citric acid cycle, cytochrome oxidase: oxidative capacity). The enzyme activities were followed in different parts of the white muscle of non-spawning female rainbow trout from May, four months after their first spawning, until December, at second spawning. Samples were taken from white epaxial muscle along the lateral line, on the dorsum, and in between. Samples were also taken from red muscle of non-spawning fish. The isoforms of myosin heavy chains (MyHC) were electrophoretically identified on 6% SDS-PAGE gel to study possible changes in contractile properties of the muscle.Transformation from the non-spawning to spawning phase was associated with dramatic changes in the activity of the enzymes studied in white muscle: glycolytic capacity decreased to less than half, whereas oxidative metabolism increased about two- to four-fold in all areas. Significant quantitative differences in enzyme activities were found between the three epaxial muscle areas: in the non-spawning fish lateral line samples differed from those taken in the other two areas, whereas in spawning fish the dorsal sample difered from the other two. No difference in the expression of MyHC-isoforms was found between spawning and non-spawning fish. Co-expression of both slow and fast isoforms was found in single fibres isolated from red muscle.The results show that the energy metabolism in white muscle of domestic rainbow trout is altered during spawning; i.e., the metabolism becomes increasingly aerobic, with an increased capacity for fatty acid utilization, concomitant with phenotypic changes associated with sexual maturation. These changes are especially pronounced in ventral, superficially located fibres.     

Influence of growth hormone on the hepatic mixed function oxidase and transferase systems of rainbow trout

The effect of GH treatment on hepatic cytochrome P450 content, aryl hydrocarbon hydroxylase (AHH), aminopyrine-N-demethylase (AND), testosterone hydroxylase, testosterone 5- and 5-reductase, UDP-glucuronyl transferase (UDPGT) and glutathione S-transferase (GST) activities in immature rainbow trout were investigated. Hepatic cytochrome P450 content, AHH and GST activities were measured in both GH implanted and GH injected animals whereas other activities were assayed in GH implanted trout only.GH implants significantly decreased cytochrome P450 content at 15 days compared to the control but no significant effect was observed at 15 or 30 d when GH was injected biweekly. In both cases, AHH activity was significantly decreased by GH treatment compared to the control whereas GST remained unchanged. Compared to the control, GH implanted fish exhibited a pronounced inhibition of AND, a decreased 6 and 16-testosterone hydroxylation, an inhibition of UDPGT with testosterone as substrate and an enhanced 17-testosterone oxidation.     

Cytochrome P-450-dependent metabolism in isolated liver cells from fed and starved rainbow trout,Salmo gairdneri

The cytochrome P-450 content in liver cells from rainbow trout was not affected by starvation for 12 weeks whereas the rate of cytochrome P-450-dependent deethylation of 7-ethoxycoumarin in liver cells from 6 or 12 weeks starved fish was 60% of the rate in fed fish. Treatment of fish with -naphthoflavone increased the 7-ethoxycoumarin metabolism several-fold in both starved and fed fish.Optimal cytochrome P-450 monooxygenation in liver cells from fed or starved fish was not affected by addition of glucose or 2-bromooctanoate, an inhibitor of fatty acid -oxidation which is the main source of metabolic fuel in trout liver. The cellular content of NADPH, an obligatory cofactor for cytochrome P-450 monooxygenation, was not affected by addition of substrate to cytochrome P-450, inhibition of fatty acid -oxidation or inhibition of the oxidative phosphorylation. This indicates a great capacity of rainbow trout liver cells to retain high NADPH/NADP⁺ ratios. These results suggest that the cytochrome P-450 mediated metabolism of xenobiotics in liver cells from fed or starved trout is not limited by the availability of reducing equivalents.     

Oxidative metabolism in a teleost,Anabas testudineus Bloch: effect of testosterone and estradiol-17β on hepatic enzyme activities

Testosterone (T) administration to maleAnabas testudineus significantly stimulated the activities of cytochrome oxidase, -glycerophosphate dehydrogenase (-GPDH) and Mg²⁺ adenosine triphosphatase (Mg²⁺ ATPase) and inhibited lactate dehydrogenase (LDH), cytosolic and mitochondrial malate dehydrogenases (MDH). The activities of succinate dehydrogenase (SDH), glucose-6-phosphate dehydrogenase (G-6-PDH) and catalase were unaffected by testosterone treatment. Administration of estradiol-17 (E2) in female fish, significantly stimulated cytochrome oxidase activity, inhibited Mg²⁺ ATPase, SDH, catalase and cytosolic and mitochondrial MDH activity, and was without effect on other enzymes studied.The simultaneous injections of actinomycin D or chloramphenicol and T or E2 prevented the hormonal influence on hepatic enzyme activities. The present study demonstrates that inA. testudineus sex steroids influence hepatic oxidative metabolism by a mechanism sensitive to the action of inhibitors of protein synthesis.     

Does the aerobic capacity of fish muscle change with growth rates?

To ascertain whether growth rate modifies the oxidative capacity of fish white muscle, we examined the effects of individual growth rate on the activities of four mitochondrial enzymes in white muscle of the fast growing Atlantic cod,Gadus morhua. Growth rates were individually monitored in cod held at three acclimation temperatures during experiments repeated in four seasons. The size dependence of citrate synthase (CS), cytochrome C oxidase (CCO) and β-hydroxyacyl CoA dehydrogenase (HOAD) activities was established using wild cod ranging from 115 to 17,350 g. Given their negative allometry, CS and CCO activities in the experimental cod were corrected to those expected for a 1.2 kg animal. HOAD activities did not change with size. The specific activities of CCO and CS were positively correlated with growth rate. However, for both enzymes, season explained more of the variability than growth rate or temperature. Season was the only factor to significantly affect the activity of HOAD, while temperature and season interacted to determine glutamate dehydrogenase activity. CS activity was positively correlated with the initial condition of the cod, which differed among the seasons. The other enzymes did not show this relationship. The independent changes of these enzymes suggest that mitochondria undergo qualitative modifications with changes in growth rate, season and size. Although growth rate and the activities of CCO and CS are positively correlated, the activity of the mitochondrial enzymes is more affected by size, physical condition and season.     

Catalytic activity and immunochemical quantification of hepatic cytochrome P-450 in β-naphthoflavone and isosafrol treated rainbow trout (Oncorhynchus mykiss)

In addition to catalytical assays, immunochemical techniques have recently been employed to measure induction of the cytochrome P-450 (P450) monooxygenase system in fish with polyaromatic hydrocarbons (PAH). In the present study, polyclonal antibodies were raised against rainbow trout P450IA1. Levels of rainbow trout P450IA1 determined using protein blotting- and ELISA procedures were compared with levels of 7-ethoxyresorufin-O-deethylase (7-EROD) activity in liver microsomes from rainbow trout. These comparisons showed that values of P450A1 were positively correlated (r=0.99 and r=0.97) with 7-EROD activities. In addition, the effects of isosafrol (ISF) or -naphthoflavone (NF) treatments on P450 levels in rainbow trout liver were investigated using immunochemical and catalytical methods. ISF treatment induced 7-EROD activity as well as 7-methoxycoumarin-O-demethylase-, 7-ethoxycoumarin-O-deethylase-, 7-propoxy-coumarin-O-depropylase and 7-butoxycoumarin-O-debutylase activities, although to a lesser extent, compared with the NF treatment. In contrast, immunochemical quantification of rainbow trout P450IA1 protein revealed a slightly different pattern. ISF appeared to be a weak inducer of P450IA1 in rainbow trout compared with NF. In addition, the degree of inhibition of 7-alkoxycoumarin-O-dealkylase activities in ISF microsomes differed from that measured in control- and NF microsomes. The discrepancies between catalytic and immunochemical estimates of rainbow trout P450IA1 in ISF treated fish in addition to differencs between specific inhibitory pattern by specific polyclonal antibodies raised against rainbow trout P450IA1, indicate that important differences exists between the responses induced by NF- and ISF treatments in the rainbow trout liver.Part of this work was presented at the 6th International Conference on Biochemistry and Biophysics of Cytochrome P-450, Vienna, Austria, July 3–8, 1988.     

Summertime and early autumn activity of some enzymes in the carbohydrate and fatty acid metabolism of the crucian carp

In June, July, and September the activities of five enzymes involved in the carbohydrate and fatty acid metabolism, namely phosphofructokinase (PFK), lactate dehydrogenase (LDH), citrate synthase (CS), cytochrome oxidase (cyt ox) and 3-hydroxy-acyl-CoA dehydrogenase (HAD), were measured in the heart, liver, red muscle, white muscle, and gill lamellae of crucian carp (Carassius carassius). LDH activity was measured in both reducing (LDHr) and oxidizing (LDHo) directions.The PFK activity in red and white muscle increased significantly between July and September indicating a preparation to winter anoxia by an increased glycylytic capacity in these organs. The HAD activity of the liver had increased significantly (by more than 50%) by September, also a preparation to winter anoxia as HAD is used in the reversed -oxidation (chain elongation of fatty acids). The LDHr and cyt ox activities in the heart and white muscle were highest in July. This shows that both the anaerobic and aerobic capacities are elevated in mid-summer when water temperature is high and oxygen concentration of the water could fluctuate greatly. The LDHo and CS activities in gill lamellae were lowest in July. The results show that the metabolism of crucian carp is under seasonal influence and that a preparation to winter hypoxia/anoxia could be detected in September.     

Cortisol does not affect hepatic α- and β-adrenoceptor properties in rainbow trout (Oncorhynchus mykiss)

Distribution and function of hepatic - and -adrenoceptors were examined in rainbow trout (Oncorhynchus mykiss) injected with slow release hydrogenated coconut oil implants alone (sham) or containing cortisol. - and -Adrenoceptors were assayed on purified hepatic membranes 10–14 days post-implantation using ³H-prazosin () and ³H-CGP (). At 10–14 days, plasma cortisol values were significantly elevated to approximately 220 compared with 35.0 ng ml^-1 in cortisol implanted vs. sham trout. No significant differences were found between any of the experimental groups for either the affinity (K_d) or maximal number of binding sites (B_max) for either receptor type. Epinephrine significantly stimulated glucose release from hepatocytes isolated from sham injected trout, but not from cortisol-treated fish. Epinephrine-induced glucose release was blocked by both - and -antagonists. These studies do not support the hypothesis that rainbow trout exposed to chronic cortisol alter properties of hepatic adrenoceptors.     

Morphometrics and ultrastructure of myocardial tissue in Notothenioid fishes

Antarctic fish of the family Channichthyidae (Icefishes) lack the respiratory pigments haemoglobin and myoglobin. The morphometrics and ultrastructure of the ventricular myocardium of a benthic icefish,Chaenocephalus aceratus has been compared with that of a red-blooded Notothenioid fish,Notothenia neglecta, of similar habit.The mass of ventricular muscle as a percentage of bodyweight is 3 times greater in adultC. aceratus (0.32%) thanN. neglecta (0.11%). Myoglobin concentration in the ventricle ofN. neglecta, 20 nmoles/g, is comparable to that of temperate teleosts with similar activity patterns. The volume and surface densities of mitochondria are 41.5% and 0.32 m^–1 for Icefish and 25% and 0.15 m^–1 forN. neglecta, Cytochrome oxidase activities are similar in the two tissues whilst the volume density of myofibrils is higher forN. neglecta (47%) thanC. aceratus (29.9%).The proliferation of mitochondria in the myocardium of Icefish will reduce the diffusion path-length for oxygen between ventricular lumen and the outer mitochondrial membrane and may compensate for the absence of myoglobin.     

The role of cAMP in regulating the β-adrenergic response of rainbow trout (Oncorhynchus mykiss) red blood cells

The -adrenergic response of teleost red blood cells (RBCs) enables the fish to maintain or even enhance the oxygen affinity of haemoglobin during various stress situations. The role of CAMP in the pronounced -adrenergic response of hypoxic rainbow trout RBCs was studied. Rainbow trout RBCs were incubated with three different -agonists (noradrenaline, adrenaline and isoproterenol, 10^–9 - 10^–4 M) at two oxygen tensions (P_{O 2}, 155 and 8 mmHg), and thereafter cAMP accumulation and cellular water content were measured.The cAMP concentration of non-stimulated trout RBCs was ca. 1200 nmol/kg dw. Of the three -agonists used, isoproterenol was the most effective in formation of cAMP, followed by noradrenaline and adrenaline. Oxygen tension affected the accumulation of cAMP in two ways. At physiological catecholamine levels (1–100 nM) there was either no difference between normoxic and hypoxic cells or a slight increase in the normoxic ones. At high catecholamine concentrations the accumulation of cAMP was greater in the hypoxic than in the normoxic cells. Oxygen tension also affected the magnitude of cell swelling but had no effect on the catecholamine concentrations causing half-maximal swelling (EC₅₀-values). The results indicate that, at physiological catecholamine levels, the -adrenergic response of rainbow trout RBCs is mainly regulated on the level of the Na⁺/H⁺ exchange.     

Seasonal variation of muscle metabolic organization in rainbow trout (Oncorhynchus mykiss)

This study examined how muscle metabolic organization varied during an annual cycle in which rainbow trout (Oncorhynchus mykiss) were held in outdoor holding ponds in which they were exposed to natural changes in temperature (range 0.2 to 15.6°C) and photoperiod. We examined the activities of glycolytic and mitochondrial enzymes in red and white muscle to evaluate whether trout enhance their capacity for lipid and carbohydrate oxidation during cold-acclimization. When assayed at habitat temperature, the enzyme activities generally increased in spring to reach a maximum in summer followed by a decrease in the fall. This led to significantly higher activities at warm than cold periods for all enzymes measured in red muscle and all but one in white muscle. The activities at 10°C provided little evidence for compensatory adjustments of aerobic capacity. Particularly in red muscle, enzyme levels at 10°C were generally lower during cold than warm periods. The variation of enzyme activities throughout the cycle was not due to changes in protein concentration, as the same responses were observed when activities were expressed per g wet mass or per mg protein. Although the aerobic capacity did not increase with cold-acclimatization, the relative capacity for lipid oxidation was higher in winter than in summer trout. In contrast, the relative capacity for aerobic glycolysis was higher in summer than in winter trout. Thus, the metabolic capacities of trout muscle undergo seasonal reorganization.     

Anatomic and metabolic responses to thermal acclimation in the ninespine stickleback,Pungitius pungitius

Male ninespine sticklebacks, Pungitius pungitius, acclimated to 3°C have higher activities of mitochondrial enzymes in their axial muscles than males acclimated to 20°C. Phosphofructokinase and pyruvate kinase activities tended to be higher in cold than warm acclimated males. For females, warm acclimation tended to decrease only mitochondrial enzyme activities. As thermal acclimation did not change the physical condition and most anatomic parameters of the sticklebacks, the enzymatic changes do not seem due to mobilization of somatic reserves. Field acclimatization to warm temperatures led to a marked decrease in physical condition in both males and females. This decrease in physical condition could largely be attributed to atrophy of the carcass mass. Spring males had higher activities of phosphofructokinase, citrate synthase and cytochrome oxidase in the axial muscle than summer males. Again, females showed a less marked response. These data suggest that environmental temperature is a major determinant of muscle aerobic capacity, at least for male ninespine sticklebacks. Thus, these northern temperate zone fish retain the capacity for thermal compensation, much like their temperate zone counterparts.     

The cytochrome P450 system of Atlantic salmon (Salmo salar): II. Variations in hepatic catalytic activities and isozyme patterns during an annual reproductive cycle

A group of Atlantic salmon (Salmo salar) was followed through their first year of maturation and spawning. At monthly intervals, starting with juvenile fish in December, 5–7 fish of each sex were killed, and liver and plasma were sampled. The last sampling point was of spawning fish in November a year later. Variables in the cytochrome P450 (P450) system were studied in hepatic microsomes, and estradiol 17 was measured in the plasma of females to assess the maturational status. The P450 1A1-mediated 7-ethoxyresorufin O-deethylase (EROD) started at high levels in winter, but decreased to non-detectable activities in pre-spawning females. Decreases, but not to the same extent, were also observed during this period in total cytochrome P450, cytochrome b₅, NADPH-cytochrome P450 reductase, and in the content of two immunochemically determined P450 isozymes. At the same time, LSI levels increased in maturing females (starting in July), and GSI levels increased in both sexes (starting in May). Sex specific differences were observed in pre-spawning fish in September and October, with levels of total P450, b₅, NADPH-cytochrome P450 reductase, EROD and P450 isozymes significantly lower in females. At the same time, plasma estradiol-17 levels reached peak values in females. The results point to the important role of sex steroids such as estradiol-17 as major factors in the regulation of final sexual maturation. However, this study also indicates that there may be estradiol-17 independent events of equal importance in the early stages of gonadal maturation that may involve the P450 system. The changes observed in the P450 system (as a major drug and steroid metabolizing system) of Atlantic salmon during sexual maturation may be of importance both in the endogenous transduction of hormonal signals, and as a pharmacological basis for designing therapeutic treatment of diseases in the aquaculture industry.Parts of this work were presented at the 5th International Symposium on Responses of Marine Organisms to Pollutants, April 1989 in Plymouth, United Kingdom (Larsen and Goks\/oyr 1989).     

Cytochrome P-450 mediated O-dealkylation of 7-alkoxycoumarins in liver microsomes from rainbow trout (Salmo gairdneri)

Evidence has recently been presented for variation in the inducibility of various 7-alkoxycoumarin-O-dealkylase activities in liver microsomes from a number of mammalian species by -naphthoflavone (NF). In the present study we have investigated the inducibility of hepatic microsomal 7-methoxycoumarin-O-demethylase, 7-ethoxycoumarin-O-deethylase, 7-propoxycoumarin-O-depropylase and 7-butoxycoumarin-O-debutylase activities in rainbow trout by NF. O-demethylase activity was increased approximately 17-fold, O-deethylase and O-depropylase activities approximately 9-fold and O-debutylase activity approximately 25-fold. The kinetics of the various hepatic microsomal 7-alkoxycoumarin-O-dealkylase activities were investigated in control and NF-treated rainbow trout. The O-demethylase-, O-depropylase- and O-debutylase activities exhibited monophasic Michaelis-Menten kinetics in liver microsomes from both control and NF-treated rainbow trout, whereas the O-deethylase activity exhibited biphasic Michaelis-Menten kinetics in control liver microsomes and monophasic Michaelis-Menten kinetics in liver microsomes from NF-treated rainbow trout.     

Plasticity of the properties of mitochondria from rainbow trout red muscle with seasonal acclimatization

Cold-acclimation of rainbow trout brings only limited changes in muscle metabolic capacities, but marked modifications in membrane composition. Thus, we examined whether the functional properties of mitochondria from trout red muscle were modified by seasonal temperature acclimatization. Mitochondria from fall-acclimatized trout had higher maximal capacities (state 3 rates) for the oxidation of pyruvate and acyl carnitines at 12 and 20 °C than mitochondria isolated from summer-acclimatized trout. For these substrates, the increased oxidative capacity completely compensated for the seasonal drop in temperature. Pyruvate and palmitoyl carnitine were consistently the preferred substrates, while decanoyl and octanoyl carnitine were oxidized at higher rates than glutamine, particularly in fall trout. State 4 rates of oxygen uptake (obtained when all ADP has been converted to ATP) differed less among substrates, but varied seasonally. State 4 rates at 12 and 20 °C were higher in mitochondria isolated from fall than summer trout. At low temperatures, the Q₁₀ of both maximal and state 4 rates of substrate oxidation tended to be higher for mitochondria from fall trout. The apparent Arrhenius activation energy (E_a) for mitochondrial pyruvate oxidation was higher in fall than summer trout whereas the E_as for palmitoyl carnitine and decanoyl carnitine oxidation did not change. The fatty acids of mitochondrial phospholipids from fall trout were more polyunsaturated than those from summer trout, with 12% more double bonds occurring than in summer trout, suggesting that membrane restructuring may be involved in the observed compensatory responses.     

Effects of food deprivation on white muscle energy reserves in rainbow trout (Oncorhynchus mykiss): the relationships with body size and temperature

The relationship between body size and the depletion of white muscle metabolites (e.g., PCr, ATP, glucose and glycogen) was examined in two sizes (30 or 700 g) of rainbow trout deprived of food for one, four or seven days at either 5 or 15 °C. Following 7 days of food deprivation at 15 °C, the levels of muscle glycogen decreased by approximately 50% in small fish relative to control values (i.e., day 1). In comparison, small fish acclimated to cold temperatures did not exhibit a significant reduction of muscle glycogen over the seven day fasting regime. In contrast to small fish, the levels of white muscle glycogen in large fish remained unchanged after food deprivation, regardless of acclimation temperature. A seven day deprivation of food also resulted in a 50% depletion of white muscle glucose concentrations in small and large fish acclimated to warm temperatures, but there were no significant changes in this variable in fish acclimated to cold temperatures. In contrast to the negative effects of food deprivation on white muscle glycogen and glucose levels, the concentrations of white muscle PCr and ATP were not greatly affected by food deprivation under any of the experimental conditions. Taken together, these results clearly show that food deprivation can have an important influence on the storage of energy metabolites for anaerobic energy production, particularly in small fish at warm temperatures. In the future, it may be very important to consider the physiological effects of short-term food deprivation when interpreting results from studies in which fish have been fasted prior to treatments such as exercise.     

The effects of hypoxia,in vivo,on red blood cell β-adrenoceptors in the rainbow trout,Oncorhynchus mykiss

Rainbow trout, Oncorhynchus mykiss, were exposed to 48h of environmental hypoxia (water partial pressure of oxygen = 8.0 kPa) at either 5 or 15°C. Blood was sampled during hypoxia via a dorsal aorta cannula to measure arterial blood partial pressure of oxygen and plasma catecholamine concentrations. After 48h, the number (B_max) and dissociation constant (K_d) of red blood cell surface -adrenoceptors were determined using a radioligand-displacement binding assay. At 5°C, plasma catecholamine levels were elevated at 24h whereas at 15°C, levels were elevated at 48h. At either temperature, following 48h of hypoxia, there was no change in B_max or K_d of red blood cell surface -adrenoceptors, compared to normoxic control fish. This study demonstrates that chronic exposure to moderate hypoxia does not affect the number or affinity of cell surface -adrenoceptors on trout red blood cells.     

Role of exogenous glutathione in teleost fish and its effects on antioxidant defense responses in rainbow trout exposed to 3,3',4,4'-tetrachlorobiphenyl

Intraperitoneal injection of glutathione (GSH) is shown to effectively increase tissue glutathione content in two teleost species, the rainbow trout and the American eel. GSH uptake into teleost tissues is a distinct piscine feature of GSH biochemistry compared with mammalian species where little or not uptake occurs of exogenously added GSH. Enhancement and depletion of tissue GSH status was used as a tool to examine the effects of altered GSH content on other cellular antioxidants in 3,3,4,4-tetrachlorobiphenyl (TCB) exposed rainbow trout. -Tocopherol content was unchanged and thiobarbituric acid reactive substances (TBARS), an indicator of lipid peroxidation, changed little in liver and muscle of trout exposed to TCB and the various GSH treatments. However, -tocopherol levels in muscle and superoxide dismutase activities in the various tissues were significantly higher in controls and TCB-saline treated fish than in GSH deficient and lipoate supplemented TCB exposed trout. GSH status thus appears to affect some cellular antioxidant defenses in TCB exposed trout.     

Changes in mitochondrial fatty acid composition following temperature acclimation of carp and their possible effects on FoF1-ATPase activity

Carp undergo temperature acclimation of respiratory function by altering mitochondrial ATP synthase (F_oF₁-ATPase) both quantitatively and qualitatively (Itoi et al. 2003). To address such acclimation temperature-dependent changes of F_oF₁-ATPase activity, we investigated in this study the correlation between the fatty acid composition and F_oF₁-ATPase activity in fast muscle of thermally acclimated carp. The quantities of saturated fatty acids of mitochondria from carp acclimated to 10 °C were significantly lower than those of carp acclimated to 30 °C. While mono- and poly-unsaturated fatty acids tended to increase with cold acclimation of carp, the molar concentration of 16:0 aldehyde in mitochondria from the 10 °C-acclimated carp were less than those from the 30 °C-acclimated fish. The specific activities of F_oF₁-ATPase in the 10 °C- and 30 °C-acclimated fish mitochondria were calculated to be 167±22 and 56±10 nmol/min ⋅ mg mitochondrial protein, respectively, the difference being significant at P<0.005. Taken together, the increase in F_oF₁-ATPase activity in fast muscle mitochondria of carp after cold temperature acclimation may be closely related to the increase of unsaturated fatty acids in mitochondria. Abbreviations: BSA - bovine serum albumin; DHA - docosahexaenoic acid; EGTA - ethyleneglycol bis(2-aminoethylether)tetraacetic acid; EPA - eicosapentaenoic acid; F_oF₁-ATPase - mitochondrial ATP synthase; α-F₁-ATPase - F_oF₁-ATPase α-subunit; β-F₁-ATPase - F_oF₁-ATPase β-subunit; HEPES - 2-[4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid; SDS - sodium dodecyl sulfate; SDS-PAGE - SDS-polyacrylamide gel electrophoresis. This revised version was published online in July 2006 with corrections to the Cover Date.     

Involvement of gonadal steroids in final oocyte maturation of white perch (Morone americana) and white bass (M. chrysops): in vivo and in vitro studies

Plasma estradiol-17 (E₂), testosterone (T), 17,20-dihydroxy-4-pregnen-3-one (DHP) and 17,20,21-tri-hydroxy-4-pregnen-3-one (20-S) levels were measured by radioimmunoassay (RIA) in white perch (Morone americana) and white bass (M. chrysops) that were induced to undergo final oocyte maturation (FOM) with human chorionic gonadotropin (hCG). Plasma DHP levels increased in females of both species in association with oocyte germinal vesicle migration (GVM) and germinal vesicle breakdown (GVBD) and decreased thereafter. Plasma 20-S levels also increased with oocyte GVM in white bass, but were several-fold lower than DHP levels. Circulating E₂ and T levels were greatest during GVM and GVBD in both species and decreased to low levels during oocyte hydration and ovulation. Follicles from white perch and white bass which received a priming injection of hCG in vivo, produced both DHP and 20-S in vitro after exposure to hCG and their oocytes underwent GVBD. Ovarian incubates from unprimed fish of either species produced only E₂ and T and their oocytes did not complete GVBD. Oocytes from unprimed bass, but not perch, matured when follicles were exposed to hCG in vitro. Both trilostane and cycloheximide blocked in vitro production of DHP and 20-S and oocyte GVBD by white perch follices. DHP and 20-S were equipotent inducers of FOM in the GVBD bioassay. None of several other structurally-related steroids tested were effective within a physiological range of concentrations. These results indicate a role for DHP and 20-S in the control of FOM in white perch and white bass.