NAD+-linked isocitrate dehydrogenase in fish tissues

NAD⁺-linked isocitrate dehydrogenase was found in the brain, heart, gills, kidney, liver and muscle of trout, and in the liver and muscle of eel. A complex homogenization buffer containing 1 mM ADP, 5 mM MgSO₄, 5 mM citrate and 40% glycerol is required for retrieval of significant amounts of stable enzyme. The highest activities were found in brain of trout and the lowest in white muscle of trout and eel. The enzyme was partially purified from frozen trout heart to a final activity of 0.04 M/min/mg protein, and the kinetic properties of this partially purified enzyme were studied. The enzyme requires either Mn²⁺ or Mg²⁺ for activity, higher activities being observed with Mn²⁺. Saturation kinetics for DL-isocitrate were sigmoidal, apparent S_0·5=8.2±0.6 mM and n_H=1.8±0.2, in the absence of ADP, changing to hyperbolic, apparent S_0·5=1.4±0.3 mM and n_H=1.0, with 1 mM ADP added. Citrate and Ca²⁺ were found to activate the enzyme to a small extent. NADH strongly inhibited the enzyme, I₅₀=3.7±0.5 M. ATP was also found to be an inhibitor, I₅₀=7.2±1.4 mM. These properties are consistent with the role of the enzyme as a major control site of the tricarboxylic acid cycle.     

Effects of ambient ion concentrations on gill ATPases in fresh water eel,Anguilla anguilla

Branchial activities of Na⁺,K⁺-ATPase (ouabain sensitive), Mg²⁺ ATPase (ouabain insensitive) and kinetic analysis of high and low affinity Ca²⁺ ATPase were measured inAnguilla anguilla that had been acclimated to demineralized water (DW, Ca < 10 M), freshwater (FW, Ca = 2 mM), and Low calcium freshwater (L-Ca, Ca = 0.9 mM). Na⁺,K⁺-ATPase activity decreased while ouabain insensitive activity increased when ambient Ca²⁺ decreased. Two kinetic forms of Ca²⁺ ATPase could be resolved in each environmental condition. The stimulation coefficients of both sites or enzymes were not affected by ambient Ca²⁺ concentrations. The maximal velocity of both the high and the low affinity Ca²⁺ ATPase was increased when external Ca²⁺ was decreased during acclimation. The low affinity Ca²⁺ ATPase and the Mg²⁺ stimulated enzyme could be a non specific enzyme accepting either Ca²⁺ or Mg²⁺. Results are compared with previous results in the literature and in relation to the branchial morphology and ionic exchanges in fish.     

Purification and physicochemical properties of deoxyribonuclease from pyloric caeca of Atlantic cod (Gadus morhuaL.)

A deoxyribonuclease (DNase) of pancreatic origin has been purified from extracts of the pyloric caeca from Atlantic cod (Gadus morhua L.). The crude extract was prepared by mincing frozen caeca tissue in equal volumes of buffer. The enzyme was isolated from the supernatant after streptomycin sulfate precipitation and centrifugation. The purification scheme further included chromatography on Q-Sepharose Fast Flow and hydroxyapatite columns. Affinity adsorption chromatography of the hydroxyapatite fraction on 8-(6-aminohexyl)-amino-5′-AMP-Sepharose, revealed an apparently homogeneous protein with molecular weight of 35,000 Da as judged by NaDodSO₄-PAGE. In sum a 644-fold enzymatic enrichment and 3.5% total enzyme recovery was achieved. The cod enzyme resembles DNase I-type enzymes with an alkaline pH activity optimum and shows dependency for Mg²⁺. The pI of the enzyme is 6.5 as determined by isoelectric focusing and DNase-zymography. Our findings suggest that the nuclease is a member of the cod's digestive enzymes secreted from the connective tissue surrounding the caeca.     

The role of Ca2+ and Na+ membrane transport in brook trout (Salvelinus fontinalis) spermatozoa motility

The role of environmental ion composition and osmolality in Ca²⁺ signaled activation was assessed in spermatozoa of brook trout Salvelinus fontinalis. Milt from ten mature males was obtained by abdominal massage. Spermatozoa motility was evaluated in 0, 100, and 300 mOsm/kg NaCl or sucrose solutions, buffered by 10 mM Tris–HCl pH 8.5. For investigation of spermatozoa reaction to external Ca²⁺ concentration, 2 mM ethylene glycol tetraacetic acid (EGTA) was added to the activation media as a calcium ions chelator. For investigation of the effect of external Na⁺ concentration in conditions of low external Ca²⁺, 100 µM amiloride was added to the EGTA-containing solutions as a Na⁺ transport blocker. Low motility was observed in sucrose (Na⁺ free) solutions containing 2 mM EGTA but not in Na⁺ solutions containing 2 mM EGTA. Addition of amiloride led to significantly increased motility (P < 0.05) compared with sucrose (Na⁺ free) solutions containing 2 mM EGTA. We conclude that Na⁺ transport in Ca²⁺-free solutions plays a regulatory role in brook trout spermatozoa activation. The influence of competitive Na⁺ and Ca²⁺ transport on the control of spermatozoa activation requires further study with respect to its application for improvement of artificial activation and storage media.     

Ionoregulatory and respiratory responses of brown trout,Salmo trutta,exposed to lethal and sublethal aluminium in acidic soft waters

Soft water acclimated (Ca²⁺ 0.02 mM; Na⁺ 0.03 mM; K⁺ 0.01 mM; pH 7.0), cannulated brown trout (Salmo trutta) were exposed to various pH and aluminium (Al) regimes (pH 7.0, pH 5.0, pH 5.0 plus Al: 50, 25, and 12.5 g l^–1) for up to 5 days in order to determine (i) the sublethal concentration of Al at pH 5.0 for this species (ii) their ionoregulatory and respiratory status. No mortality or physiological disturbances were evident at pH 7.0 or pH 5.0. All trout died within 48 h at pH 5.0 in the presence of Al at 50 g l^–1 and 67% died over the 5 day period at pH 5.0 in the presence of Al at 25 g l^–1. Fish at these lethal Al concentrations showed significant decreases in arterial blood oxygen content (C_aO₂) but no changes in plasma osmolarity or the concentrations of plasma Na⁺, K⁺ and Cl^–. Physiological disturbance was more marked at the 50 g l^–1 Al concentration. The surviving fish at 25 g l^–1 showed few signs of physiological recovery while continually exposed to this regime. No fish died during the exposure to water of pH 5.0 containing 12.5 g l^–1 Al, but physiological disturbance was still apparent. These sublethally-stressed trout showed a transient decline in the plasma concentrations of Na⁺ and Cl^–1. Although C_aO₂ decreased, recovery was evident. The data suggest that in the brown trout, environmental Al concentration is as important as pH and calcium concentration in determining the physiological status of the fish.     

Effects of Zn2+ on Ca2+ uptake by mitochondria and endoplasmic reticulum in permeabilized tilapia gill cells

An intracellular ATP-dependent Ca²⁺ pumping mechanism, distinct from mitochondrial Ca²⁺ accumulation, was identified within tilapia gill cells. Cell suspensions treated with 0.003% saponin, which selectively permeabilizes the plasma membrane, were used to characterize the Ca²⁺ sequentering mechanisms as endoplasmic reticulum and mitochondria and to determine the effect of Zn²⁺ on their Ca²⁺ storing activity. Of the Ca²⁺ taken up by the endoplasmic reticulum, 80% was released by IP₃ (10 mol l^–1). The Ca²⁺ pump of the endoplasmic reticulum was 2.5 times less sensitive to Zn²⁺ (IC₅₀=0.05 nmol l^–1) than was the mitochondrial uptake mechanism (IC₅₀=0.20 nmol l^–1). The results indicate that Ca²⁺ is stored predominantly within the endoplasmic reticulum at 0.1 mol l^–1 and that this storing capacity is seriously attenuated by namomolar concentrations Zn²⁺.     

Mitochondrial NAD(P)-malic enzyme from herring skeletal muscle

Mitochondrial NAD(P)-dependent malic enzyme [EC 1.1.1.39, L-malate: NAD⁺ oxidoreductase (decarboxylating)] was purified from herring skeletal muscle to a specific activity of 8.2 mol/min/mg. The purification procedure involved chromatography on DEAE-cellulose, Red Agarose and a Sephacryl S-300 with a final recovery of 38% of enzyme activity. This enzyme catalyzes the oxidative decarboxylation of malate in the presence of either NAD or NADP in the presence of Mn²⁺. Some kinetic characteristics of this enzyme were determined. The pH optimum of activity is 7.0. ATP was shown to be a competitive inhibitor with malate. The inhibition by ATP displayed hyperbolic competitive kinetics with a K_i (ATP) of 0.28 mM in the presence of NAD and 0.75 mM in the presence of NADP. Fumarate reversed ATP inhibition.In vivo, regulation of NAD(P)-dependent malic enzyme might respond to changing levels of mitochondrial ATP and fumarate with the enzyme undergoing kinetic activation by an increase in the concentration of mitochondrial fumarate which could reverse enzyme inhibition by ATP.     

Increased winter temperature did not affect completion of smolting in Atlantic salmon

Four groups of 1+ year-old Atlantic salmon, Salmo salar, pre-smolts were reared under various temperature regimes: constant 10 °C from November onwards; ambient temperature until either 15 December or 1 February, then 10 °C; or ambient temperature throughout (control; 2–3 °C November–March). From 20 May onwards, temperature in all groups was ambient, increasing from 10 °C to 17 °C in late July. Rearing temperature had no significant effect on either the timing of completion of smolting, or the duration of the smolt-window. Mean gill Na⁺-K⁺ ATPase activity was not significantly affected by temperature regime; it increased in all groups from < 2.0 mol Pi mg protein^–1 h^–1 (units) in January to 5–7 units in mid-April, then back to < 2.0 units in July. Survival in 96 h, 37) salinity (S) tolerance tests was similar in all groups, increasing from < 10% in early March, to > 90% from mid-April to mid-June, then decreasing to < 20% by early July. Increased winter temperature significantly increased growth and condition factor compared with the control, but during April–May all four groups exhibited similar temporary decreases in condition factor in association with the completion of smolting.     

Mechanism of Cd<Superscript>2+</Superscript> on DNA cleavage and Ca<Superscript>2+</Superscript> on DNA repair in liver of silver crucian carp

The subject of acute injury, apoptosis and canceration of animals induced by heavy metal ions has been one of the hotspots studied worldwide. However, the exact molecular mechanism of Cd-induced carcinogenicity remains largely unclear, and how to relieve the toxicity in vivo has rarely been reported. For this paper, we have investigated the mechanism of Cd²⁺ on DNA cleavage and Ca²⁺ on DNA repair in the liver of silver crucian carp (Carassius auratus gibelio) by agarose gel electrophoresis methods and by estimating biochemical indexes. Our results show that Cd²⁺ induces the classical laddering degradation of DNA in vivo and that DNA cleavage is repaired after injection with Ca²⁺ under various Cd²⁺ concentrations. DNA cleavage caused by Cd²⁺ is due to the activation of deoxyribonuclease (DNase) and the accumulation of reactive oxygen species (ROS). Furthermore, Cd²⁺ destroys the antioxidant system, which diminishes the activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD), causing an increase of the lipid peroxidation (LPO) level, respectively. However, after the liver is injected with Ca²⁺ under various Cd²⁺ concentrations, the DNase activity, the ROS generating rate and the LPO level are obviously reduced, the activities of SOD, CAT, and POD are greatly increased. At the same time, ROS production and removal recoves its balance. The results show that Ca²⁺ can relieve the toxicity of Cd²⁺ in silver crucian carp.     

Purification and characterization of chymotrypsin from viscera of vermiculated sailfin catfish, Pterygoplichthys disjunctivus, Weber, 1991

Pterygoplichthys disjunctivus viscera chymotrypsin was purified by fractionation with ammonium sulfate (30–70 % saturation), gel filtration, affinity, and ion exchange chromatography. Chymotrypsin molecular weight was approximately 29 kDa according to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), shown a single band in zymogram. Electrofocusing study suggested being an anionic enzyme (pI ≈ 3.9), exhibiting maximal activity at pH 9 and 50 °C, using Suc-Ala-Ala-Pro-Phe-p-nitroanilide (SAAPNA) as substrate. Enzyme was effectively inhibited by phenyl methyl sulfonyl fluoride (PMSF) (99 %), and N-tosyl-l-phenylalanine chloromethyl ketone (TPCK) (94 %). Enzyme activity was affected by the following ions in decreasing order: Hg²⁺, Fe²⁺, Cu²⁺, Li¹⁺, Mg²⁺, K¹⁺, Mn²⁺, while Ca²⁺ had no effect. Chymotrypsin activity decreased continuously as NaCl concentration increased (from 0 to 30 %). K _m and V _max values were 0.72 ± 1.4 mM and 1.15 ± 0.06 μmol/min/mg of protein, respectively (SAAPNA as substrate). Results suggest the enzyme has a potential application where low processing temperatures are needed, such as in fish sauce production.     

The effects of epibionts and predators on the growth and mortality rates of Argopecten purpuratus cultures in southern Chile

Epibiont and predator effects on the growth and mortality of the northern scallop Argopecten purpuratus were evaluated in cultures in southern Chile.The most common epibionts were bryozoans, hydrozoans and algae in both study sites, Metri Bay (41°36; 72°43W) and Chidhuapi Channel (41°49S; 73°05W).After 12 months of culture in pearl nets and lantern nets, the scallops size did not show statistically significant differences in cultures with and without epibionts in both study sites nor were there differences in the growth either with or without the presence of predators (decapod crustaceans). The growth rate was higher in Chidhuapi Channel than in Metri Bay.Mortality was concentrated in the initial phase of culture in pearl nets. During the culture phase in lantern nets, the mortality rate was lower than 3%. The mortality rate in scallops with epibionts was higher than when these were removed in the culture phase in pearl nets. In Metri Bay the mortality rate with predators was higher than without predators. Epibionts and predators did not affect mortality in the culture phase in lantern nets.Epibionts and predation are important factors in the early mortality of scallops and therefore in the success of culture. Epibiosis, however, is not important in scallop growth in southern Chile. This is related to the composition of the epibionts and to the low temperatures which probably limit the growth of algae and invertebrates.     

Trypsin Enzyme from Viscera of Common Kilka (Clupeonella cultriventris caspia): Purification,Characterization, and Its Compatibility with Oxidants and Surfactants

The purification of trypsin from the common kilka (Clupeonella cultriventris caspia) viscera (pyloric caeca) resulted in a 28.3-fold increase and 12% yield by ammonium sulfate precipitation (30–60%), Sephadex G-75, and DEAE-cellulose chromatography. Trypsin showed a molecular weight of 23.2 kDa and appeared as a single band on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), native-PAGE, and zymography. The trypsin had optimal activity at pH 8.0 and 60°C for the hydrolysis of α-N-benzoyl-DL-arginine-ρ-nitroanilide hydrochloride (BAPNA) substrate. Trypsin was stable up to 50°C and at pH range of 7.0–10.0. Activity was significantly inhibited by soybean trypsin inhibitor (SBTI) and N-ρ-tosyl-L-lysine-chloromethylketone (TLCK) inhibitors (p < 0.05). The enzyme was relatively stable toward oxidizing agents, retaining 59.7 and 98.0% of its initial activity after 1 h incubation in the presence of 15% H₂O₂ and 1% sodium perborate, respectively. Trypsin was significantly activated by surfactants and Ca²⁺, Mg²⁺, and Mn²⁺ and inactivated by Fe²⁺, Zn²⁺, Cu²⁺, Al³⁺, Ba²⁺, and Co²⁺ (p < 0.05). Nevertheless, Na⁺ and K⁺ had no significant effect on trypsin activity (p > 0.05). The purified trypsin showed significantly higher catalytic efficiency (k_cat/K_m) than porcine pancreatic trypsin against BAPNA and N-α-p-Tosyl-L-arginine methyl ester hydrochloride (TAME) substrates (p < 0.05).     

The effect of seawater flow and temperature on metamorphosis and postlarval development in great scallop

Variations in growth and survival of hatchery-reared post-metamorphicjuveniles of great scallop Pecten maximus prompted anexamination of settlement and postlarval development. The effects ofseawater flow and temperature on great scallop metamorphosis andpostlarvae were studied over a 4–5 week period. In allexperiments, and regardless of environmental conditions, great scallopmetamorphosed after a 2–3 week period with values of 35 to70%. Subsequently, spat numbers increased slightly. Spatmortality generally occurred from the third week onward and reachedlevels as high as 30% by the fifth week under standardconditions. At 20 °C, however, 60% mortality levels wererecorded. Differences in spat growth rate, ranging from 37 to 45 mday^–1, were noticed at different seawater flow ratesbut no clear tendency could be discerned. Temperature affected spatgrowth with an increase in size from 24 m day^–1 at15 °C to 35 m day^–1 at 18 °C. Conversely,growth was suppressed at 20 °C (14 m day^–1).For optimal metamorphosis and postlarval development in great scallop, aseawater flow of 4.3 L h^–1 per sieve and a temperatureof 15 °C are recommended.     

Catfish electroreceptor organ functioning during five days exposure to different calcium environments

Catfish, Ictalurus melas, were pre-adapted to artificial tap water with 1.2 mM Ca²⁺ for two weeks, and subsequently transferred to artificial tap water with 0.6, 1.2, or 2.2 mM Ca²⁺ for one week in order to investigate the effect of the environmental Ca²⁺-concentration on stimulus encoding and the frequency characteristics in ampullary electroreceptor organs. Within 30 minutes after transfer, the spontaneous activity of the primary afferents, as well as gain and phase of the stimulus induced responses changed transiently corresponding to the Ca²⁺-concentration. One day after transfer the Ca²⁺-induced changes of the spontaneous activity had disappeared as well as the differences between the sensitivities at frequencies of 2, 8, 12, 16 and 20 Hz in 0.6 and 1.2 mM Ca²⁺, whereas at 16 and 20 Hz in 2.2 mM Ca²⁺ the sensitivity was still elevated. The Ca²⁺-induced phase shift was strongly frequency dependent. At 2 Hz no Ca²⁺ effect on the phase was observed, whereas at 12, 16 and 20 Hz significant effects could be demonstrated up till three days after transfer. The latency was not affected by the transfer.The Ca²⁺-induced effects on the primary afferent spontaneous activity are probably related to a Na⁺/Ca²⁺-exchanger at the basal faces of the receptor cells. The frequency dependent effects on gain and phase are concluded to relate to properties of the apical membrane, most likely to Ca-dependent K-channels. These findings further support the concept that ampullary electroreceptor might serve as chemoreceptor organs.     

解淀粉芽孢杆菌HZ-1510壳聚糖酶基因的克隆、重组表达及其活性

为了研究壳聚糖酶的水解活性,实验进行了解淀粉芽孢杆菌HZ-1510壳聚糖酶基因编码序列的克隆,并构建了谷胱甘肽转移酶(GST)-壳聚糖酶融合蛋白表达质粒,在大肠杆菌中表达后提取、纯化得到重组蛋白,并通过生物信息学对该蛋白的信号肽、三维结构等进行了分析,最后以胶态壳聚糖溶液为底物研究了该重组壳聚糖酶的活性。结果显示,该壳聚糖酶基因的ORF长为837 bp,编码279个氨基酸,分子量为31.45 ku。在其氨基末端具有信号肽,切割点位于36和37位氨基酸之间。氨基酸序列同源性分析表明该壳聚糖酶属于GH46家族糖苷水解酶。酶的水解活性最适温度约为55°C,最适pH为5.5。而金属离子Fe3+、Ag+、Cu~(2+)、Ba~(2+)和K+对其水解活性都起抑制作用,但是Mn~(2+)、Ca~(2+)和Mg~(2+)对其活性起增强作用。0.1 mmol/L的Zn~(2+)对壳聚糖酶的活性起增强作用,2.0 mmol/L的Zn~(2+)对壳聚糖酶的活性起抑制作用。本实验研究了解淀粉芽孢杆菌HZ-1510壳聚糖酶在不同条件下的水解活性,为壳聚糖酶的工业应用奠定了理论基础。     

Variations in growth in haemoglobin genotypes of Atlantic cod

In the present paper are described the growth properties of three different haemoglobin genotypes of juvenile Atlantic cod (Gadus morhua) reared at 7, 10, 13 and 16 °C. In addition one group was reared under temperature steps i.e. moved successively from 16 to 13 and 10 °C. The genotype Hb-I(2/2)displayed the overall highest growth rate in the temperature range 13–16 °C, whereas the Hb- I(1/1)genotype showed the highest overall growth at the lowest temperature (7 °C). Accordingly, we found a significant interaction between genotype and temperature. The differences in growth were largest when cod were reared under the temperature step regime where the Hb-I(2/2)genotype displayed 17 and 24% higher growth than Hb-I(1/1)and Hb-I(1/2),respectively. Optimal temperature for growth (T_opt.G) varied between the genotypes with the genotype Hb-I(1/2)displaying the highest (mean ± SE) T_opt.G (14.5 ± 1:0.8 °C) and Hb-I(1/1)the lowest (12.5 ± 0.2 °C). The biological significance of this link between biochemical genetic variation and physiological properties might be the influences on growth pattern, ultimate size and age at first maturity.     

Feeding Electivity Indices in Surgeonfish (Acanthuridae) of The Florida Keys

Three species of surgeonfish Acanthurus coeruleus (blue tang) (Block and Schneider), A. bahianus (ocean surgeonfish) (Castelnau), and A. chirurgus (doctorfish) (Block) were captured in the waters of the Florida Keys (24°03N, 81°40W). In total, 39 of these fish were captured between March and December of 1999. Items found in the stomachs of these fish were identified and analyzed for percent occurrence. These values were compared to percent occurrence values of the same items from random bottom transects taken at the point of capture to quantify any forage selectivity or avoidance behavior of these fish. A. bahianus selected for sand and chlorophytes and avoided phaeophytes. A. chirurgus also showed selection for sand and chlorophytes, while 58% of those sampled selected for phaeophytes and 42% avoided them. A. coeruleus avoided sand and selected for rhodophytes and chlorophytes. Phaeophytes were also selected for in 80% of A. coeruleus sampled. Understanding food preferences of free-ranging Florida surgeonfish may provide insight to improve nutritional management of captive herbivorous reef fish.     

Interaction of low density lipoproteins with liver cells in rainbow trout

Liver is the main catabolic tissue for low density lipoprotein in rainbow trout (Gjøen and Berg 1992). We have investigated the interaction of LDL with isolated trout liver cells and liver membranes. ¹²⁵I-TC labelled trout LDL bound to isolated trout liver cells in a time dependent and saturable manner with an apparant K_d of 20.1 g/ml, suggesting the existence of a specific binding site on the surface of these cells. The binding was Ca²⁺ dependent assessed by the 50% reduction obtained by 5 mM EDTA. Saturable binding to isolated trout liver membranes could also be demonstrated, but with lower affinity as compared to intact cells. Degradation of ¹²⁵I-TC-LDL in hepatocytes was also saturable as degradation could be inhibited about 60% by a 100 fold surplus of unlabelled LDL. The rate of degradation increased with temperature up to 20°C. Both cell association (binding + uptake) and degradation were reduced down to 20% of control in the presence of microtubular and lysosomal inhibitors. Hepatic catabolism of trout LDL therefore seems to depend on receptormediated endocytosis, followed by lysosomal degradation.Abbreviations TC tyramine cellobiose - LDL low density lipoproteins - MeLDL methylated low density lipoproteins - VLDL very low density lipoproteins - HDL high density lipoproteins - VTG vitellogenin - EDTA ethylenediamine tetraacetic acid - PBS phospate buffered saline - SDS-PAGE sodium dodecyl sulphatepolyacrylamide gel electrophoresis - DMPC L--phosphatidylcholine-dimyristoyl     

The effect of K+, Ca2+, and Mg2+ on sperm motility in the perch,Perca fluviatilis

This study investigated the effect of 0.25–5 mM K⁺, Ca²⁺, and Mg²⁺ on sperm motility in the perch, Perca fluviatilis. In 75 mM NaCl, the used motility-activating solution, motility rate, and swimming velocity decreased within the first 4 min after activation, and the rate of locally motile sperm increased. Thereafter, the motility parameters remained constant for periods >20 min. Based on the decrease in sperm motility, two types of semen samples could be distinguished. Semen samples of type I retained a high motility rate of >65 % after 20 min, and the rate of locally motile sperm was <20 %. In semen samples of type II, the motility rate decreased to values <30 % after 20 min, and the rate of locally motile sperm exceeded >50 %. Ca²⁺ and Mg²⁺ concentrations of 0.25–0.5 mM had no effect on the sperm motility parameters 10 s after activation, while 0.25 mM K⁺ increased the swimming velocity. K⁺, Ca²⁺, and Mg²⁺ concentrations ≥1.5 mM had suppressive effects on the sperm motility 10 s after activation. No differences were found between the two semen types. Twenty minutes after activation, type I semen was not affected by the tested cations. On the contrary, 0.25–2.5 mM K⁺, 0.25 mM Mg²⁺, and 0.25–2.5 mM Ca²⁺ significantly increased the sperm motility rate and/or sperm velocity of type II semen. Therefore, supplementation of saline solution with cations might stabilize the motility of perch sperm, which can be a benefit for experimental purposes and for specific handling procedures in aquaculture.     

Actions of epinephrine on the contractility of fast and slow skeletal muscle fibres in teleosts

Fast and slow muscle fibers were isolated from the myotomes of atlantic cod (Gadus morhua L.) and sculpin (Myoxocephalus scorpius L.). Epinephrine was found to have no effect on twitch or sub-tetanic contractions in fast muscle fibres. Isoprenaline (10^–6M) had no effect on the contractility of slow muscle fibres. In contrast, epinephrine elicited a dose-dependent decrease in the half-time for twitch relaxation (t_1/2r), and in most cases a decrease in twitch amplitude. The maximum decrease in t_1/2r was around 5–20% of control values (at 10^–6M epinephrine), with a half maximal response at about 30 nmol l^–1. Responses to epinephrine were unaffected by propranolol and reversed by phentolamine, consistent with the stimulation of -adrenoreceptors. 10^–6M epinephrine produced a rise in cAMP levels from 1.8 to 3.1 pmol mg dry wt^–1 in cod slow fibres. However, the cellular mechanism underlying the action of epinephrine is unclear since forskolin, a potent activator of adenylate cyclase activity, where it has been investigated, was found to increase not decrease twitch duration and amplitude. The responses of fast and slow fibres to epinephrine and its antagonists were similar in summer (13°C) and winter acclimatized (5–6°C) sculpin.It is suggested that epinephrine may act to modulate the active state of slow muscle fibres at high cruising speeds and thereby increase swimming performance.