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Effect of Triiodothyronine on the Activity and Sensitivity of Glycosidases to Heavy Metals (Cu, Zn, and Pb) in Juvenile Blue Bream Ballerus Ballerus (L.)

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Effect of Triiodothyronine on the Activity and Sensitivity of Glycosidases to Heavy Metals (Cu, Zn, and Pb) in Juvenile Blue Bream Ballerus Ballerus (L.) See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/319860350 Effect of triiodothyronine on the activity and sensitivity of glycosidases to heavy metals (Cu, Zn, and Pb) in juvenile blue bream Ballerus ballerus (L.) Article in Inland Water Biology · July 2017 DOI: 10.1134/S1995082917030063 CITATION READS 1 40 4 authors: Andrey Filippov Aleksey Bolotovskiy Russian Academy of Sciences Russian Academy of Sciences 39 PUBLICATIONS 113 CITATIONS 13 PUBLICATIONS 55 CITATIONS SEE PROFILE SEE PROFILE Boris Levin Irina Leonidovna Golovanova Russian Academy of Sciences Russian Academy of Sciences 198 PUBLICATIONS 766 CITATIONS 105 PUBLICATIONS 473 CITATIONS SEE PROFILE SEE PROFILE Some of the authors of this publication are also working on these related projects: Spatial distribution and qualitative composition of POPs in freshwaters of Russia View project Method in morphometric studies of Phenetic Diversity View project All content following this page was uploaded by Aleksey Bolotovskiy on 16 October 2017. The user has requested enhancement of the downloaded file. ISSN 1995-0829, Inland Water Biology, 2017, Vol. 10, No. 3, pp. 305–307. © Pleiades Publishing, Ltd., 2017. Original Russian Text © A. Filippov, A.A. Bolotovskiy, B.A. Levin, I.L. Golovanova, 2017, published in Biologiya Vnutrennykh Vod, 2017, No. 3, pp. 62–65. ECOLOGICAL PHYSIOLOGY AND BIOCHEMISTRY OF HYDROBIONTS Effect of Triiodothyronine on the Activity and Sensitivity of Glycosidases to Heavy Metals (Cu, Zn, and Pb) in Juvenile Blue Bream Ballerus ballerus (L.) A. Filippov, A. A. Bolotovskiy, B. A. Levin, and I. L. Golovanova* Papanin Institute for Biology of Inland Waters, Russian Academy of Sciences, Borok, 152742 Russia *е-mail: [email protected] Received October 14, 2016 Abstract⎯The influence of exogenous triiodothyronine (0.25 ng/mL) on the activity glycosidases (maltase, amylolytic activity) and their sensitivity to Cu, Zn and Pb ions (25 mg/L) has been studied in the whole organ- ism of juvenile blue bream Ballerus ballerus (L.). We have found that the treatment of fish with triiodothy- ronine resulted in an increase in amylolytic activity. Maltase activity is not affected. In addition, the exposure to exogenous triiodothyronine results in a decrease in the sensitivity of glycosidase to the in vitro action of the ions of biogenic metals Cu and Zn and in an increase in the sensitivity of the maltase to the action of Cu, Zn, and Pb ions. Keywords: Cyprinidae, blue bream, digestive glycosidases, triiodothyronine, heavy metals, Cu, Zn, Pb DOI: 10.1134/S1995082917030063 INTRODUCTION It was revealed that Т3 may affect the activity of gly- cosidases in the intestine of roach Rutilus rutilus (L.) Thyroid hormones (THs) are one of the most (fam. Cyprinidae) [3]. Another cyprinid species, blue important groups of hormones that start to function in bream Ballerus ballerus (L.), exhibits the lowest level fish since early ontogenesis. These hormones play an of Т3 in the blood plasma among all studied fish spe- important role in the metabolic and morphogenetic cies; i.e., this species is TH-deficient [9]. In this processes (including metamorphosis), in regulating respect, the study on the influence of Т3 on the activity the reproductive system, and in the regeneration of of glycosidases in the TH-deficient species is espe- certain organs and tissues [8, 11, 13]. It is believed that cially interesting. thyroxin is a prohormone, while 3,5,3'- triiodothy- The goal of this paper is to study the effect of ronine (Т3) is a more active form of thyroid hormones formed by the deiodination of thyroxin [8]. triiodothyronine on the activities of glycosidases and their sensitivity to the effects of Cu, Zn, and Pb ions in Aquatic organisms are subject to a range of anthro- juvenile blue bream Ballerus ballerus (L.). pogenic factors, such as salts of heavy metals strongly affecting natural ecosystems. Copper, zinc, and lead are the priority pollutants of ambient waters [5]. MATERIALS AND METHODS Unlike lead, copper and zinc are essential micronutri- Juvenile blue breams were exposed to T3. The gam- ents participating in many biochemical reactions. etes were obtained from one female and three males However, in large quantities the latter two elements are caught by seine net in the Rybinsk Reservoir near toxic to aquatic life. In the areas affected by techno- Borok (Yaroslavl oblast, Russia) in early May 2015. genic pollution, concentrations of these metals reach Fish eggs were fertilized by the dry method, placed on several milligrams per 1 L of water [4]. Entering organ- pieces of glass, and incubated for 8 days in a 3-L plastic ism with water and food, copper, zinc, and lead may container at a water temperature of 11.2–15.8°С. Fol- affect fish digestive enzymes both directly and indi- lowing hatching, the larvae were transferred into a 60-L rectly [1, 7]. The lysosomal hydrolases of tissues of aquarium. The larvae and fries were maintained at food organisms (prey) may considerably contribute to natural regimes of temperature fluctuations from 16.5 the processes of digestion in fish [10]. This is why it is to 24.1°С and illumination (12 h light : 12 h dark). necessary to study the effects of heavy metals not only After switching to exogenous feeding, the fries were fed upon the digestive hydrolases of a consumer, but also ad libitum with Sera Micron dry food for aquarium upon the enzymes in the prey organisms. fishes, live Artemia sp. nauplii, and frozen Chironomus sp. 305 306 FILIPPOV et al. Table 1. Activity of glycosidases in the whole organism of solutions of substrates (18 g/L starch and 50 mM/L juvenile blue bream of control and test (Т3) groups upon maltose) were prepared using the same Ringer’s solu- exposure to Cu, Zn and Pb ions in vitro tion. The solutions of enzymatically active prepara- Metal ions Activity of glycosidases, μM/(g min) tions and substrates were incubated for 20–30 min at 20°С, рН 7.4 at constant mixing. To study the effects (25 mg/L) amylolytic maltase of heavy metals, the homogenates were preliminarily incubated with salts of copper (CuSO4 ⋅ 5H2O), zinc 1.34± 0.01а 2.16± 0.05а Absence (ZnSO4 ⋅ 7H2O), and lead (Pb(NO3)2 for 1 h. The con- 1.53± 0.07а 2.14± 0.02а centrations of Cu, Zn, and Pb ions, as calculated by ± c ± а the content of metal in salt, was 25.0 mg/L. The amy- Cu 0.92 0.02 1.93 0.15 c b lolytic activity reflecting the sum activities of the 1.09± 0.02 1.98± 0.05 enzymes hydrolyzing starch, α-amylase EC 3.2.1.1, ± b ± a glucoamylase EC 3.2.1.3, and maltase EC 3.2.1.20, Zn 1.13 0.03 2.23 0.03 a b was assessed by the gain of hexoses following modified 1.39± 0.04 1.95± 0.03 Nelson’s technique [6]. To determine the maltase ± b ± a Pb 1.17 0.03 2.31 0.04 activity by the glucose oxidase method, the Photoglu- 1.25± 0.04b 1.89± 0.02c cosa (OOO Impact, Russia) test for clinical biochem- istry was used. The optical density was read on a Means and errors of means are given; different letters superscript indicate statistically significant differences between the parame- Lambda 25 (PerkinElmer, United States) spectropho- ters in a row (ANOVA, LSD-test), p < 0.05. Control is on top; test tometer at wavelengths of 670 nm (amylolytic activity) is on bottom. and 505 nm (maltase activity). The activities of enzymes were expressed in micromolar of the reaction product formed for 1 min incubation per 1 g wet tissue once a day. At the age of 22 days post fertilization, the weight (μM/(g ⋅ min)). fish were split into two groups, with 35 individuals in each group. The fish of the first group were maintained The results are given as means and errors of means in an aquarium with clean water (control); those of the (M ± m). The normality of distribution of studied second group were in an aquarium with triiodothy- parameters was assessed using Shapiro–Wilk test. Sta- ronine (Т ) at a concentration of 0.25 ng/mL (test vari- tistical significance of differences was assessed using 3 single-factor analysis (ANOVA followed by LSD-test) ant). The conditions (temperature, illumination, and at ≤ 0.05. feeding) were the same for both groups. Every day, p 1/3–1/2 of the water volume in the aquarium was changed, with the Т3 concentration kept constant. RESULTS AND DISCUSSION The duration of the experiment was 85 days (fish age of 107 days after fertilization). No differences in either length or weight in juvenile fish of two experimental groups were revealed by the After the termination of exposure, Т3 was extracted end of experiment. Body length and weight in control from the whole fish (12 individuals from control and test fish were 29.8 ± 0.7 mm and 0.30 ± 0.02 g, respec- groups) tissue homogenates using lysing buffer (0.1 M tively; in the test group it was 31.8 ± 1.4 mm and 0.41 ± phosphate buffered saline pH 7.4, 0.1% Triton X-100, 0.05, respectively. By the end of experiment, the con- 1 mM propylthiouracil – Sigma Aldrich). The sam- centration of Т3 in the tissues of test-group fish was ple-to-buffer ratio was 1 : 1 by weight. After homoge- higher by a factor of three (0.75 ± 0.05 ng/mL) than in nization and centrifuging (for 30 min at 10000 g) of the control (0.25 ± 0.05 ng/mL).
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