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Purification and Properties of White Muscle Lactate Dehydrogenase from the Anoxia-Tolerant Turtle, the Red-Eared Slider, Trachemys scripta elegans.

ABSTRACT: Lactate dehydrogenase (LDH; E.C. 1.1.1.27) is a crucial enzyme involved in energy metabolism in muscle, facilitating the production of ATP via glycolysis during oxygen deprivation by recycling NAD(+). The present study investigated purified LDH from the muscle of 20 h anoxic and normoxic T. s. elegans, and LDH from anoxic muscle showed a significantly lower (47%) K m for L-lactate and a higher V max value than the normoxic form. Several lines of evidence indicated that LDH was converted to a low phosphate form under anoxia: (a) stimulation of endogenously present protein phosphatases decreased the K m of L-lactate of control LDH to anoxic levels, whereas (b) stimulation of kinases increased the K m of L-lactate of anoxic LDH to normoxic levels, and (c) dot blot analysis shows significantly less serine (78%) and threonine (58%) phosphorylation in anoxic muscle LDH as compared to normoxic LDH. The physiological consequence of anoxia-induced LDH dephosphorylation appears to be an increase in LDH activity to promote the reduction of pyruvate in muscle tissue, converting the glycolytic end product to lactate to maintain a prolonged glycolytic flux under energy-stressed anoxic conditions.

SUBMITTER: Dawson NJ

PROVIDER: S-EPMC3594981 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Background:How species colonize new environments is still a fundamental question in ecology and evolution, assessable by evaluating range characteristics of invasive species. Here we propose a model approach to evaluate environmental conditions and species features to predict niche changes in non-equilibrium contexts. It incorporates potentially range-limiting processes (fundamental niche), hence allowing for better predictions of range shifts, differentiation of analog and non-analog conditions between the native and non-native (invaded) ranges, and identification of environmental conditions not currently available but likely in the future. We apply our approach with the worldwide invasive slider-turtle Trachemys scripta. Methods:We estimated the native and non-native realized niches of T. scripta and built its fundamental niche based on key features of the turtle's temperature physiological tolerance limits and survival-associated factors. We next estimated response functions adjusted to the physiological predictor variables and estimated habitat suitability values, followed by a comprehensive set of analyses and simulations to compare the environmental conditions occupied by T. scripta (at its native and non-native ranges). Results:Climatic space analysis showed that the T. scripta's non-native realized niche is 28.6% greater than the native one. Response curves showed that it does not use its entire range of temperature tolerances (density curves for native: 5.3-23.7 °C and non-native: 1.7-28.4 °C ranges). Whether considering the mean temperature of the warmest or the coldest quarter, it occupies a wider range of temperatures along its non-native distribution. Results of the response curves for worldwide (global) and across Mexico (regional) comparisons showed it occupies analog and non-analog conditions between its native and invaded ranges, exhibiting also unoccupied suitable climatic conditions. Discussion:We demonstrate that T. scripta occupies a wider subset of its fundamental niche along its non-native range (within its physiological tolerances), revealing that the species observed niche shift corresponds to a different subset of its fundamental niche (niche unfilling). We also identified suitable environmental conditions, globally and regionally, where the slider turtle could potentially invade. Our approach allows to accurately predict niche changes in novel or non-equilibrium contexts, which can improve our understanding about ecological aspects and geographic range boundaries in current and potential invasions.

Project description:Aquatic animals have developed various mechanisms to live in either hyperionic or hypoionic environments, and, as such, not many species are capable of surviving in both. The red-eared slider turtle, Trachemys scripta elegans, a well-known freshwater species, has recently been found to invade and inhabit brackish water. Herein, we focus on some of the metabolic adaptations that are required to survive and cope with salinity stress. The regulation of the adenosine monophosphate (AMP)-activated protein kinase (AMPK), a main cellular "energy sensor," and its influence on lipid metabolism were evaluated with a comparison of three groups of turtles: controls in freshwater, and turtles held in water of either 5‰ salinity (S5) or 15‰ salinity (S15) with sampling at 6, 24, and 48 h and 30 days of exposure. When subjected to elevated salinities of 5 or 15‰, AMPK mRNA levels and AMPK enzyme activity increased strongly. In addition, the high expression of the peroxisome proliferator activated receptor-α (PPARα) transcription factor that, in turn, facilitated upregulation of target genes including carnitine palmitoyltransferase (CPT) and acyl-CoA oxidase (ACO). Furthermore, the expression of transcription factors involved in lipid synthesis such as the carbohydrate-responsive element-binding protein (ChREBP) and sterol regulatory element-binding protein 1c (SREBP-1c) was inhibited, and two of their target genes, acetyl-CoA carboxylase (ACC) and fatty acid synthase (FAS), were significantly decreased. Moreover, exposure to saline environments also increased plasma triglyceride (TG) content. Interestingly, the content of low-density lipoprotein cholesterol (LDL-C) and total cholesterol (TC) in plasma was markedly higher than the control in the S15 group after 30 days, which indicated that lipid metabolism was disrupted by chronic exposure to high salinity. These findings demonstrate that activation of AMPK might regulate lipid metabolism in response to salinity stress through the inhibition of lipid synthesis and promotion of lipid oxidation in the liver of T. s. elegans. This may be an important component of the observed salinity tolerance of these turtles that allow for invasion of brackish waters.

Dataset Information

Purification and Properties of White Muscle Lactate Dehydrogenase from the Anoxia-Tolerant Turtle, the Red-Eared Slider, Trachemys scripta elegans.

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