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Deuterium-Depleted Water Has Stimulating Effects on Long-Term

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Deuterium-Depleted Water Has Stimulating Effects on Long-Term G Model NSL 30844 1–5 ARTICLE IN PRESS Neuroscience Letters xxx (2014) xxx–xxx Contents lists available at ScienceDirect Neuroscience Letters jo urnal homepage: www.elsevier.com/locate/neulet 1 Deuterium-depleted water has stimulating effects on long-term 2 memory in rats a,∗ b,c b a c,d 3 Q1 Cristian Mladin , Alin Ciobica , Radu Lefter , Alexandru Popescu , Walther Bild a 4 University of Craiova, Faculty of Chemistry, Craiova, Romania b 5 Alexandru Ioan Cuza University, 11 Carol I Blvd., 700506 Iasi, Romania c 6 Center of Biomedical Research of the Romanian Academy, Iasi Branch, Romania d 7 Gr. T. Popa University of Medicine and Pharmacy, 16 Universitatii Street, 700115 Iasi, Romania 8 9 h i g h l i g h t s 10 • 11 Deuterium-depleted water resulted in no significant changes in Y-maze. • 12 Reference memory errors were decreased in the DDW group in the radial 8 arms-maze. • 13 Time to finish the radial was reduced in DDW, compared to distilled water-controls. 14 2615 a r t i c l e i n f o a b s t r a c t 16 17 Article history: Deuterium-depleted water (DDW) is a water which has a 6–7-fold less concentration of the naturally 18 Received 24 May 2014 occurring deuterium (20–25 ppm vs. 150 ppm). While administrated for a longer period, it may reduce 19 Accepted 18 September 2014 the concentration of deuterium throughout the body, thus activating cellular mechanisms which are 20 Available online xxx depending on protons (channels, pumps, enzyme proteins). The aim of the present work was to study, 21 for the first time in our knowledge, the possible influence of deuterium-depleted water (DDW) chronic 22 Keywords: administration in normal Wistar rats, as compared to a control group which received distilled water, 23 Deuterium-depleted water on spatial working memory and the locomotor activity (as studied through Y-maze) or both short-term 24 Working memory and long-term spatial memory (assed in radial 8 arms-maze task). Our results presented here showed no 25 Spatial memory significant modifications in terms of spatial working memory (assessed through spontaneous alternation percentage) and locomotor activity (expressed through the number of arm entries) in Y-maze, as a result of DDW ingestion. Also, no significant differences between the DDW and control group were found in terms of the number of working memory errors in the eight-arm radial maze, as a parameter of short-term memory. Still, we observed a significant decrease for the number of reference memory errors in the DDW rats. In this way, we could speculate that the administration of DDW may generate an improvement of the reference memory, as an index of long-term memory. Thus, we can reach the conclusion that the change between the deuterium/hydrogen balance may have important consequences for the mechanisms that govern long-term memory, as showed here especially in the behavioral parameters from the eight-arm radial maze task. © 2014 Published by Elsevier Ireland Ltd. 27 1. Introduction phenomenon called kinetic isotope fractionation [33]. This phe- 33 nomenon represents the separation of molecules during physical 34 28 Water (H2O), the most common compound on Earth and the processes like evaporation or liquefaction according to their molec- 35 29 absolute necessity for life, does not have an identical isotopic ular mass. Heavier molecules, like in the case of water, heavy 36 30 composition throughout all liquid bodies on Earth. Differences water (D2O) or semiheavy water (HDO) will accumulate in the 37 31 may be minute, but nevertheless important from the biological lower part atmosphere, while the lighter H2O molecules will be 38 32 standpoint. Its inhomogeneous isotopic composition is due to a the first to evaporate. This phenomenon is widely used in separat- 39 ing D2O from its lighter counterparts HDO and H2O, for industrial 40 purposes. 41 ∗ It is known that the natural levels of deuterium in terrestrial 42 Corresponding author. Tel.: +40 751218264; fax: +40 232201472. E-mail address: [email protected] (C. Mladin). waters varies from 14 to 150 ppm [14], which means, grosso modo, 43 http://dx.doi.org/10.1016/j.neulet.2014.09.037 0304-3940/© 2014 Published by Elsevier Ireland Ltd. Please cite this article in press as: C. Mladin, et al., Deuterium-depleted water has stimulating effects on long-term memory in rats, Neurosci. Lett. (2014), http://dx.doi.org/10.1016/j.neulet.2014.09.037 G Model NSL 30844 1–5 ARTICLE IN PRESS 2 C. Mladin et al. / Neuroscience Letters xxx (2014) xxx–xxx 44 that one of each 6500 atoms of hydrogen composing the biological Cantacuzino Institute Bucharest, Romania, while the behavioral 104 45 structures is a deuterium one. tasks which we used were represented by Y-maze and eight-arm 105 46 When the heavier isotope of hydrogen, deuterium, was discov- radial maze (Coulbourn Instruments). The control groups received 106 47 ered, the investigation of its combinations with hydrogen to form distilled water from the same source mentioned above (Institute of 107 48 deuterated water had begun [25]. The research for establishing the Criogeny and Isotopic Research Râmnicu-Vâlcea). Distilled water 108 49 biological effects of heavy (deuterated) water was extensive. Many was used as a control according to several other studies that used 109 50 studies were made for establishing its biological actions in ani- it on behavioral evaluations, in durations varying from 7 days to 110 51 mals and plants [13]. It has been long-time observed that while the several months [7,28]. 111 52 small amounts (around 145–150 parts per million-ppm) are normal 53 in all surface waters, increased amounts of deuterium, exceeding 2.3. Experimental design 112 54 15% determine structural, metabolic and functional alterations in 55 various degrees [15]. 2.3.1. Treatment 113 56 One of the until recently unstudied byproducts of the separation The animals were randomly divided in two groups: 20 animals 114 57 process for obtaining heavy water is the so-called deuterium- for the control group and 20 for the DDW treatment. 115 58 depleted water (DDW). Initially considered a waste product, the The treatment began 21 days before the behavioral testing. 116 59 water that left the distillation columns after the extraction process Memory functions were assessed through Y-maze and eight-arm 117 60 was discarded, until studies demonstrated that it contained a deu- radial maze tasks and performed during the days 22–37 of treat- 118 61 terium content which was way below the natural levels. Further ment (Y-maze on the 22th day and radial maze in the last 2 weeks). 119 62 extraction lead to obtain water which is composed of almost only Thus, only one set of animals was used for both behavioral tasks. 120 63 H2O, with very small amounts of deuterium (around 20 ppm). The aforementioned duration of treatment was selected consid- 121 64 Compared with the biologic changes determined by the excess ering that a minimum of 21 days DDW treatment was deemed 122 65 deuterium in the water, the effects of reducing its concentration necessary in order to replace all the water in the organism and 123 66 under the normal values are less studied. There is only a small to change the hydrogen/deuterium ratio within the tissues of the 124 67 amount of data published in the literature concerning the inhibi- test animals [20]. 125 68 tion both of the growth rhythm of the fibroblast cultures and of Additionally, the investigator was blind to the type of water 126 69 the development of a tumor transplanted in mice [26], and just ingested by the rats during the behavioral tasks. 127 70 a few other publications which claim antineoplastic [9], antiaging 71 [3], cell-stimulating [5] and radiation-protecting effects [6]. 2.4. Evaluation of memory function 128 72 As the deuterium concentration difference between the intra- 73 cellular and extracellular environments seems to balance rather 2.4.1. Y-maze task 129 74 quickly, a decrease of the deuterium concentration in the cellular Short-term memory was assessed by spontaneous alternation 130 75 microenvironment should produce also a decrease of the intracel- behavior during a single session in the Y-maze task. The Y-maze 131 76 lular deuterium [24]. used in the present study consisted of three arms (35 cm long, 132 77 In this context, the aim of the present work was to study the 25 cm high, and 10 cm wide) and an equilateral triangular cen- 133 78 possible influence of DDW chronic administration in normal Wistar tral area. The maze was placed in exactly the same place during 134 79 rats, as compared to a control group which received distilled water, the procedure for all rats. In this way, the rat was placed at the 135 80 on spatial working memory and the locomotor activity (as studied end of one arm and allowed to move freely through the maze for 136 81 through Y-maze) or both short-term and long-term spatial memory 8 min. An arm entry was counted when the hind paws of the rat 137 82 (assed in radial 8 arms-maze task). were completely within the arm. Also, the maze was cleaned with 138 alcohol-free disinfectant wipes between each trial. Spontaneous 139 83 2. Materials and methods alternation behavior was defined as the entry into all three arms 140 on consecutive choices. The number of maximum spontaneous 141 84 2.1. Animals alternation behaviors was calculated as the total number of arms 142 entered minus 2 and percent spontaneous alternation was calcu- 143 85 Adult male Wistar (n = 40) rats, weighing 200–250 g at the begin- lated as (actual alternations/maximum alternations) × 100.
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