Optimization of Acetylcholinesterase and Metabolic Enzyme Activity in Multiple Fish Species

2007 ◽  
Vol 42 (2) ◽  
pp. 101-110 ◽  
Author(s):  
Eugene T. Georgiades ◽  
Douglas A. Holdway

Abstract An examination of acetylcholinesterase (AChE), citrate synthase (CS) and lactate dehydrogenase (LDH) activities in the brains of a range of fish species, occupying different trophic levels, is useful to investigate the full extent of the effects of sublethal or pulse exposures to pesticides. This investigation explores the optimization of microplate procedures for AChE, CS and LDH measurements in the brain tissue of species commonly used in laboratory investigations and those common to Canadian watersheds. A microplate assay was optimized for the measurement of AChE in the brain homogenates of seven fish species. The critical aspects of this assay requiring optimization were pH, substrate concentration and tissue dilution. Incubation with specific cholinesterase inhibitors indicated that enzymatic activity in the brain homogenates of each species was attributed to AChE only. Microplate assays were also optimized for the measurement of the metabolic enzymes, CS and LDH, in the brain homogenates of six fish species. For these assays, low interspecies variability was exhibited between optimized factors including pH, substrate, chromogen and cofactor concentrations. For each assay optimized, enzyme activities in the brain homogenates were stable for 2 to 3 hours post-thaw. Results from the present study will aid future ecotoxicological investigations of the potential impacts of AChE inhibition on neuronal glucose metabolism.

2019 ◽  
Vol 25 (33) ◽  
pp. 3519-3535 ◽  
Author(s):  
Md. Tanvir Kabir ◽  
Md. Sahab Uddin ◽  
Mst. Marium Begum ◽  
Shanmugam Thangapandiyan ◽  
Md. Sohanur Rahman ◽  
...  

: In the brain, acetylcholine (ACh) is regarded as one of the major neurotransmitters. During the advancement of Alzheimer's disease (AD) cholinergic deficits occur and this can lead to extensive cognitive dysfunction and decline. Acetylcholinesterase (AChE) remains a highly feasible target for the symptomatic improvement of AD. Acetylcholinesterase (AChE) remains a highly viable target for the symptomatic improvement in AD because cholinergic deficit is a consistent and early finding in AD. The treatment approach of inhibiting peripheral AChE for myasthenia gravis had effectively proven that AChE inhibition was a reachable therapeutic target. Subsequently tacrine, donepezil, rivastigmine, and galantamine were developed and approved for the symptomatic treatment of AD. Since then, multiple cholinesterase inhibitors (ChEIs) have been continued to be developed. These include newer ChEIs, naturally derived ChEIs, hybrids, and synthetic analogues. In this paper, we summarize the different types of ChEIs which are under development and their respective mechanisms of actions.


2001 ◽  
Vol 58 (8) ◽  
pp. 1585-1593 ◽  
Author(s):  
Jill BK Leonard ◽  
Stephen D McCormick

To evaluate the metabolic differences between Atlantic salmon (Salmo salar) parr and smolts and the effect of rearing environment, we examined metabolic enzyme activity in white muscle, liver, and heart in stream- and hatchery-reared juveniles. Spring increases in gill Na+,K+-ATPase (3.5-fold) and cardiosomatic index (37–69%) and decreases in condition factor (~17%) occurred in smolts, but not in parr. White muscle phosphofructokinase (PFK) increased during spring and was 3.6-fold higher in smolts than in parr by late spring. There were seasonal increases in liver citrate synthase (CS) (~42%), liver β-hydroxyacyl-coenzyme A dehydrogenase (HOAD) (~60%), and heart CS (~23%) and decreases in liver lactate dehydrogenase (LDH) (~28%) in parr and smolts. Activity of liver HOAD was greater in stream-reared smolts (~18%) than in parr or hatchery smolts. Heart PFK activity increased during spring in wild-reared parr and smolts, while it decreased in hatchery-reared smolts. White muscle LDH and PFK increased earlier in spring in hatchery- than in stream-reared smolts. Our results suggest that increased heart size and high white muscle PFK occur during smolting and may be adaptive for downstream and ocean migration. Hatchery- and stream-reared Atlantic salmon differ in the timing of metabolic changes during smolting, which may impact their long-term survival.


1993 ◽  
Vol 265 (6) ◽  
pp. R1344-R1350 ◽  
Author(s):  
J. M. Lawler ◽  
S. K. Powers ◽  
T. Visser ◽  
H. Van Dijk ◽  
M. J. Kordus ◽  
...  

Inhibition of metabolic enzyme activity has been associated with free radical stress in locomotor muscle with prolonged or intense exercise. However, it is not known whether such alterations with acute exercise in skeletal muscle are influenced by muscle fiber type or age. Twenty 4-mo-old and twenty 24-mo-old female Fischer-344 rats were divided at random into young exercised (YE; n = 10), old exercised (OE; n = 10), young control (YC; n = 10), and old control (OC; n = 10) groups. Animals in both YE and OE groups ran on a treadmill (10% uphill grade) for 40 min at approximately 75% of each age-group's maximal O2 consumption. Immediately after the treadmill run, white gastrocnemius (WG), red gastrocnemius (RG), and soleus (SOL) muscles were removed and quick-frozen in liquid nitrogen. Malondialdehyde was significantly increased (P < 0.05) in RG of YE vs. YC rats. Glutathione peroxidase activity was significantly elevated (P < 0.05) in the WG of YE rats. Analysis of variance revealed a significant over-all increase in superoxide dismutase activity with exercise. Activities of phosphofructokinase (PFK), citrate synthase, succinate dehydrogenase, and 3-hydroxyacyl-CoA dehydrogenase were unchanged (P > 0.05) with acute exercise in the SOL. However, PFK activity was decreased in the WG by 60% in OE but only 33% in YE, and in the RG by 41% in OE but only 21% in YE. We conclude that maximal glycolytic flux in the gastrocnemius may be adversely affected by acute exercise, and this effect was more pronounced in the 24-mo-old group.


2020 ◽  
Vol 16 ◽  
Author(s):  
Anupama Sharma ◽  
Renu Bist ◽  
Hemant Pareek

Background:: Current study evaluated the protective potential of quercetin against lindane induced toxicity in mice brain. For investigation, mice were allocated into four groups; First group was control. Second group was administered with oral dose of lindane (25 mg/kg bw) for 4 consecutive days. Third group was exposed to quercetin (40 mg/kg bw) and in fourth group, quercetin was administered 1 hour prior to the exposure of lindane. Objective:: Two major objectives were decided for study. First was to create lesions in the brain by lindane and; second was to evaluate the neuroprotective potential of quercetin. Methods:: To study oxidative responses, level of thiobarbituric acid reactive substances (TBARS), protein carbonyl content (PCC), reduced glutathione (GSH), superoxide dismutase (SOD), Catalase (CAT), and glutathione peroxidase (GPx) were measured in brain homogenates. Three key step regulating enzymes of tricarboxylic acid (TCA) cycle viz citrate synthase (CS), pyruvate dehydrogenase (PDH) and fumarase were also assayed. Results:: Lindane treatment significantly enhanced the levels of TBARS (P<0.001),PCC (P<0.001), GPx (P<0.001), SOD (P<0.05), PDH (P<0.05) and fumarase (P<0.001) in brains of mice compared to control. Meanwhile, it alleviated GSH, CAT and CS (P<0.05) activity. Conclusion:: Pretreatment with quercetin in lindane treated group not only restored, previously altered biochemical parameters after lindane treatment and also significantly improved them too which suggests that quercetin is not only invulnerable rather neuroprotective against lindane intoxication.


2021 ◽  
Vol 166 ◽  
pp. 112172
Author(s):  
Tiphaine Mille ◽  
Amaëlle Bisch ◽  
Nathalie Caill-Milly ◽  
Pierre Cresson ◽  
Jonathan Deborde ◽  
...  

1995 ◽  
Vol 269 (1) ◽  
pp. R64-R72 ◽  
Author(s):  
C. M. Bishop ◽  
P. J. Butler ◽  
S. Egginton ◽  
A. J. el Haj ◽  
G. W. Gabrielsen

Preflight development of the goslings was typified by rapid increases in the mitochondrial enzymes of the semimembranosus and heart ventricular muscles resulting in near-adult values by 3 wk of age. In contrast, aerobic capacity of the pectoralis muscle initially developed slowly but showed a rapid increase between 5 and 7 wk of age, in preparation for becoming airborne. Activities of glycolytic enzymes in the pectoralis muscle showed similar patterns of development as those found for the aerobic enzymes, except for hexokinase, which was low at all ages, indicating an adaptation for catabolism of both intracellular glycogen and plasma fatty acids in preference to plasma glucose. Muscle mass specific activity of citrate synthase in the pectoralis increased by only 33% from goslings during the first few days of flight, compared with premigratory geese. Activities of anaerobic glycolytic enzymes in the ventricles were low, but values for hexokinase, which is involved in the phosphorylation of plasma glucose, developed rapidly. Values for lactate dehydrogenase were also high, reflecting the capacity of the heart to catabolize plasma lactate. Substrate flux supplied by carnitine palmitoyltransferase and oxoglutarate dehydrogenase (OGD), in the pectoralis muscles of the premigratory geese, appears to have the smallest excess capacities to meet the requirements of sustained aerobic flight. The average maximum oxygen uptake for premigratory geese during flight, as indicated by values for OGD, is calculated to be 484 ml O2/min (or 208 ml O2.min-1.kg-1).


2018 ◽  
Vol 2018 ◽  
pp. 1-10 ◽  
Author(s):  
Martina Reutzel ◽  
Rekha Grewal ◽  
Carmina Silaidos ◽  
Jens Zotzel ◽  
Stefan Marx ◽  
...  

Aging represents a major risk factor for developing neurodegenerative diseases such as Alzheimer’s disease (AD). As components of the Mediterranean diet, olive polyphenols may play a crucial role in the prevention of AD. Since mitochondrial dysfunction acts as a final pathway in both brain aging and AD, respectively, the effects of a mixture of highly purified olive secoiridoids were tested on cognition and ATP levels in a commonly used mouse model for brain aging. Over 6 months, female NMRI mice (12 months of age) were fed with a blend containing highly purified olive secoiridoids (POS) including oleuropein, hydroxytyrosol and oleurosid standardized for 50 mg oleuropein/kg diet (equivalent to 13.75 mg POS/kg b.w.) or the study diet without POS as control. Mice aged 3 months served as young controls. Behavioral tests showed deficits in cognition in aged mice. Levels of ATP and mRNA levels of NADH-reductase, cytochrome-c-oxidase, and citrate synthase were significantly reduced in the brains of aged mice indicating mitochondrial dysfunction. Moreover, gene expression of Sirt1, CREB, Gap43, and GPx-1 was significantly reduced in the brain tissue of aged mice. POS-fed mice showed improved spatial working memory. Furthermore, POS restored brain ATP levels in aged mice which were significantly increased. Our results show that a diet rich in purified olive polyphenols has positive long-term effects on cognition and energy metabolism in the brain of aged mice.


2021 ◽  
Vol 9 ◽  
Author(s):  
Paula Evelyn Rubira Pereyra ◽  
Gustavo Hallwass ◽  
Mark Poesch ◽  
Renato Azevedo Matias Silvano

Trophic levels can be applied to describe the ecological role of organisms in food webs and assess changes in ecosystems. Stable isotopes analysis can assist in the understanding of trophic interactions and use of food resources by aquatic organisms. The local ecological knowledge (LEK) of fishers can be an alternative to advance understanding about fish trophic interactions and to construct aquatic food webs, especially in regions lacking research capacity. The objectives of this study are: to calculate the trophic levels of six fish species important to fishing by combining data from stable isotopes analysis and fishers’ LEK in two clear water rivers (Tapajós and Tocantins) in the Brazilian Amazon; to compare the trophic levels of these fish between the two methods (stable isotopes analysis and LEK) and the two rivers; and to develop diagrams representing the trophic webs of the main fish prey and predators based on fisher’s LEK. The fish species studied were Pescada (Plagioscion squamosissimus), Tucunaré (Cichla pinima), Piranha (Serrasalmus rhombeus), Aracu (Leporinus fasciatus), Charuto (Hemiodus unimaculatus), and Jaraqui (Semaprochilodus spp.). A total of 98 interviews and 63 samples for stable isotopes analysis were carried out in both rivers. The average fish trophic levels did not differ between the stable isotopes analysis and the LEK in the Tapajós, nor in the Tocantins Rivers. The overall trophic level of the studied fish species obtained through the LEK did not differ from data obtained through the stable isotopes analysis in both rivers, except for the Aracu in the Tapajós River. The main food items consumed by the fish according to fishers’ LEK did agree with fish diets as described in the biological literature. Fishers provided useful information on fish predators and feeding habits of endangered species, such as river dolphin and river otter. Collaboration with fishers through LEK studies can be a viable approach to produce reliable data on fish trophic ecology to improve fisheries management and species conservation in tropical freshwater environments and other regions with data limitations.


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