Characterization and Subcellular Localization of Leucine Aminonaphthylamidase (LAN) in Rainbow Trout (Salmo gairdneri)

1975 ◽  
Vol 32 (8) ◽  
pp. 1289-1295 ◽  
Author(s):  
G. R. Bouck ◽  
P. W. Schneider Jr. ◽  
Janet Jacobson ◽  
R. C. Ball

LAN analyses appear to have diagnostic value in fish pathobiology and studies were undertaken to determine optima for substrate concentration, pH, reaction time, temperature, and buffer ions. Citrate ion did not inhibit LAN at anticoagulant levels, but cyanide, pyrophosphate, and EDTA had an inhibitory effect. Storage of samples at —10 and 1 C resulted in small but significant reductions of LAN activity, while at room temperature enzyme activity was rapidly lost. LAN activity was distributed among liver fractions as follows: microsomes, 12%; mitochondria, 9%; cellular sap, 37%; other, 50%. Three isozymes of LAN were found. Blood plasma contained significant amounts of LAN activity which was significantly higher in cold- than in warm-acclimated fish. However, these LAN levels were comparable when their activity was extrapolated to body temperatures.

1950 ◽  
Vol 28e (3) ◽  
pp. 69-79 ◽  
Author(s):  
R. J. Rossiter ◽  
Esther Wong

Rabbit polymorphonuclear leucocytes contain an enzyme capable of hydrolyzing biosynthetic phenolphthalein mono-β-glucuronide. The concentration of the enzyme in the white cell is some 2000 times the concentration of the enzyme in the blood plasma. Under the conditions of study, the β-glucuronidase activity was proportional to the concentration of the enzyme. The effect of substrate concentration on the enzyme activity was studied and the Michaelis constant, Ks, determined. The course of the reaction was linear with time for the first 12 hr. and then fell off slightly during the next 12 hr. The optimum pH of the enzyme was 4.45 in either 0.2 M acetate or 0.2 M phthalate buffer. It was not inhibited by cyanide, azide, iodoacetate, fluoride, glycine, thiourea, urethane, arsanilic acid, acetophenone, o-cresol or m-cresol, in a final concentration of 0.01 M. The possible function of β-glucuronidase in rabbit polymorphonuclear leucocytes is discussed.


1976 ◽  
Vol 65 (1) ◽  
pp. 157-177 ◽  
Author(s):  
P. W. Webb

The fast-start (acceleration) performance of seven groups of rainbow trout from 9-6 to 38-7 cm total length was measured in response to d.c. electric shock stimuli. Two fast-start kinematic patterns, L- and S-start were observed. In L-starts the body was bent into an L or U shape and a recoil turn normally accompanied acceleration. Free manoeuvre was not possible in L-starts without loss of speed. In S-starts the body was bent into an S-shape and fish accelerated without a recoil turn. The frequency of S-starts increased with size from 0 for the smallest fish to 60–65% for the largest fish. Acceleration turns were common. The radius of smallest turn for both fast-start patterns was proportional to length (L) with an overall radius of 0–17 L. The duration of the primary acceleration stages increased with size from 0–07 s for the group of smallest fish to 0–10 s for the group of largest fish. Acceleration rates were independent of size. The overall mean maximum rate was 3438 cm/s2 and the average value to the end of the primary acceleration movements was 1562 cm/s2. The distance covered and velocity attained after a given time for fish accelerating from rest were independent of size. The results are discussed in the context of interactions between a predator and prey fish following initial approach by the predator. It is concluded that the outcome of an interaction is likely to depend on reaction times of interacting fish responding to manoeuvres initiated by the predator or prey. The prey reaction time results in the performance of the predator exceeding that of the prey at any instant. The predator reaction time and predator error in responses to unpredictable prey manoeuvre are required for prey escape. It is predicted that a predator should strike the prey within 0-1 s if the fish are initially 5–15 cm apart as reported in the literature for predator-prey interactions. These distances would be increased for non-optimal prey escape behaviour and when the prey body was more compressed or depressed than the predator.


1986 ◽  
Vol 43 (10) ◽  
pp. 2017-2024 ◽  
Author(s):  
Pierre Raymond ◽  
Gérard Leduc ◽  
Jack A. Kornblatt

The exposure of rainbow trout (Salmo gairdneri) to 0.01, 0.02 and 0.03 mg HCN∙L−1 for 20 d demonstrated the biotransformation of HCN into thiocyanate which accumulated in the blood plasma which increased the duration of exposure to cyanide. Despite its transformation into thiocyanate, there is still a sufficient quantity of cyanide to inhibit cytochrome oxidase by 60–80% within the first 24 h of exposure. This level of inhibition remained stable during the 20 d of the tests and was the same at the three concentrations tested. This observation permits the prediction of effective concentration of free HCN inside the fish. The measurements of liver glycogen levels illustrate the Pasteur effect in the rainbow trout subjected to cyanide. During cyanide exposure, liver glycogen was rapidly used, particularly at 0.03 mg HCN∙L−1 whereas at 0.01 and 0.02 mg HCN∙L−1 the glycogen levels gradually came back to normal after 7–10 d of exposure.


1980 ◽  
Vol 37 (1) ◽  
pp. 116-120 ◽  
Author(s):  
Gerald R. Bouck

The levels of soluble protein (Biuret-reactive) and levels of leucine aminonaphthylamidase (LAN, 3.4.1.1) were determined quantitatively in extracts of 10 organs from rainbow trout (Salmo gairdneri). Soluble protein had a very discrete distribution and averaged 5.2 times more concentrated in liver than in blood plasma (3.5 g/100 mL). LAN was present in the extract of every tissue except saline-washed brain. LAN activity was highest in hindgut, 36 more times more concentrated than in the plasma of healthy fish. Wasting disorders such as caused by stress, would likely increase the level of necrosis which in turn would increase the amount of LAN in plasma. I propose that a "tissue equivalent" unit of LAN to provide a means of relating increased LAN in plasma to an amount of liver destruction in vitro that would render a similar amount of LAN.Key words: plasma enzymes, stress, plasma protein, rainbow trout, leucine aminonaphthylamidase, fish physiology


1979 ◽  
Vol 36 (4) ◽  
pp. 366-376 ◽  
Author(s):  
S. A. Spigarelli ◽  
M. M. Thommes

Body temperatures of rainbow trout (Salmo gairdneri) caught from a thermal discharge into Lake Michigan were used to determine selected temperatures and to estimate acclimation temperatures of this species under field conditions. Approximately 65–75% of the variability in body temperatures was related to water temperature (direct) and fish weight (inverse). Body temperatures increased with increases in discharge temperature, but the difference between body and ambient water temperatures decreased at high water temperatures. The modal body temperature of small fish was 19 °C, the final preferred temperature predicted for rainbow trout by some lab studies. The modal body temperature of large fish was 15 °C. Estimates of acclimation temperatures indicate that the majority of rainbow trout caught in this discharge area were acclimated to plume temperatures. Estimated acclimation temperatures exceeded ambient acclimation by as much as 10 °C for individual fish while group means ranged between 2 and 6 °C over ambient acclimation. Key words: temperature selection, acclimation, thermal plume, rainbow trout, Lake Michigan, body temperature, size effect


1968 ◽  
Vol 25 (7) ◽  
pp. 1465-1474 ◽  
Author(s):  
Ulf H. M. Fagerlund ◽  
J. R. McBride ◽  
Edward M. Donaldson

Metopirone (SU 4885), an inhibitor of 11β-hydroxylation of adrenocorticosteroids, was administered intramuscularly to adult castrated sockeye salmon (Oncorhynchus nerka) and intact rainbow trout (Salmo gairdneri). Marked hypertrophy of interrenal cells pointed to an increase in the activity of the interrenal tissue of both species. Increased cortisol concentrations in trout at low dosage levels of metopirone suggested that the drug was stressful. However, decreased concentrations at higher dosage levels in trout, and decreased cortisol concentrations in all salmon indicated that the drug was also exerting a marked inhibitory effect on 11β-hydroxylation. In salmon, a repository ACTH preparation (acthar) brought about interrenal hypertrophy similar to that produced by metopirone. Metopirone also induced degranulation and hyperplasia of the cells of the palisade-like layer in the rostral region of the pars distalis in both species of fish. These cells were the only cell type of the pars distalis that were stained with lead-haematoxylin. The evidence indicates that they are the corticotrops.


1967 ◽  
Vol 24 (8) ◽  
pp. 1701-1715 ◽  
Author(s):  
T. Nakano ◽  
N. Tomlinson

In rainbow trout (Salmo gairdneri) blood plasma concentrations of adrenaline (AD) and noradrenaline (NAD), and liver and heart concentrations of AD increased in response to severe physical disturbance. Skeletal muscle and anterior kidney concentrations of AD and NAD did not change detectably. Maximum plasma concentrations observed during disturbance of the fish were 0.20–0.36 μg AD and 0.05–0.09 μg NAD/ml. These plasma concentrations decreased relatively rapidly during recovery of the fish.Plasma glucose concentrations increased in response to disturbance, the magnitude and duration of the hyperglycemia being greater in those fish with initially high liver glycogen reserves. Liver glycogen concentrations in those fish with initially high (ca. 7%) concentrations apparently decreased in response to disturbance and increased during recovery of the fish, but no change was detected in liver glycogen concentrations in fish in which they were initially low (ca. 2.5%). Heart and skeletal muscle glycogen concentrations decreased in response to disturbance and increased during recovery.In skeletal muscle, the concentration of adenosine 3′,5′-phosphate and the proportion of phosphorylase in the a form increased in response to disturbance of the fish and decreased thereafter.


1975 ◽  
Vol 10 (1) ◽  
pp. 118-125 ◽  
Author(s):  
Gérard Leduc ◽  
Kenneth K.S. Chan

Abstract The effects of cyanide on iono- and osmoregulation are soon established and last long after the period of exposure to the toxicant is over. The effects are of the order of 4 to 8 percent changes in blood plasma osmolality and chloride ions, changes which are indicative of serious physiological impairment, having costly energetic implications. The deleterious effects of cyanide were detected at concentrations as low as 0.01 mg/l HCN in water of pH 7.5. Juvenile rainbow trout, Salmo gairdneri, were first exposed to various concentrations of cyanide ranging from 0.01 to 0.037 mg/1 HCN for 28 days in fresh water in flow-through aquaria at 10?C. After this exposure period the fish together with untreated fresh water controls were transferred into 18.9 ppt salt water for 260 hours during which blood plasma osmolality and chloride were monitored. By the end of the salinity tolerance tests the poisoned fish had experienced greater loss of water than the controls but no change of chloride were observed. Upon return to fresh water for 100 hours all the fish lost chloride ions, the effect being much greater in the cyanide exposed fish. When the fish were first adapted to salt water then transferred to fresh water cyanide they also experienced loss of chloride and dilution of plasma greater than the controls did.


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