Expression of neuronal migration-related factors and the spatial learning and memory of rats prenatally exposed to tritiated water

Q4 Medicine 中华放射医学与防护杂志 Pub Date : 2019-12-25 DOI:10.3760/CMA.J.ISSN.0254-5098.2019.12.002

Yongsheng Wang, Mingming Wang

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Abstract

Objective To study the expression of neuronal migration-related factors and spatial learning and memory of rats exposed to tritiated water (HTO). Methods Hippocampal neural cells from newborn Sprague-Dawley(SD) rats at postnatal 24 h were primarily cultured in DMEM/F12 medium with 20% of fetal bovine serum for 6 days, followed by subjection to tritiated water(HTO) at concentrations of 3.7×102, 3.7×103, 3.7×104, 3.7×105, 3.7×106 Bq/ml for 24 h, respectively. Western blot and RT-qPCR were used to determine the expression levels of F-actin, α-tubulin, tau, AP2, BDNF mRNA and Reelin mRNA. 16 pregnant SD rats at embryonic (E) day 14 were randomly divided into the tested and control groups (8 rats/ each group). The tested rats were injected with body fluid of HTO (3.7×106 Bq/g) intraperitoneally, while the saline as the control. Morris water maze (MWM) was employed for the spatial learning and memory of rats. Results Compared to the control cells, HTO caused a significant downregulation of expressions of cytoskeletal proteins [F-actin (t=8.898-19.896, P<0.05), α-tubulin (t=3.261-7.900, P<0.05), tau (t=2.274-5.003, P<0.05), and MAP2 (t=2.274-5.003, P<0.05)] and mRNA of BDNF(t=3.580-19.792, P<0.05) and Reelin (t=3.240-39.692, P<0.05) in the tested neural cells in a dose-dependent manner. In addition, the escape latency of irradiated offsprings was significantly prolonged (t=-2.563, P<0.05), the time for offsprings to cross through target quadrant was markedly reduced (t=3.214, P<0.05), and the swimming time in the platform quadrant of irradiated offsprings were obviously shortened (t=3.874, P<0.05) in the MWM trial. Conclusions The results indicate that HTO irradiation in utero downregulates the expressions of neuron migration-related factors and induces brain dysfunction, which may shed a light on a mechanism of the radiation-induced brain impairment. Key words: Neuronal cell; Tritiated water; Neuronal migration related factor; Spatial learning and memory

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氚化水环境下大鼠神经迁移相关因子的表达与空间学习记忆

目的研究氚化水(HTO)对大鼠神经迁移相关因子及空间学习记忆的影响。方法将出生24 h的SD大鼠海马神经细胞先在含20%胎牛血清的DMEM/F12培养基中培养6 d，然后分别用浓度为3.7×102、3.7×103、3.7×104、3.7×105、3.7×106 Bq/ml的氚化水(HTO)培养24 h。Western blot和RT-qPCR检测F-actin、α-微管蛋白、tau、AP2、BDNF mRNA和Reelin mRNA的表达水平。选取胚胎第14天妊娠SD大鼠16只，随机分为试验组和对照组(每组8只)。实验大鼠腹腔注射HTO体液(3.7×106 Bq/g)，生理盐水作为对照组。Morris水迷宫(MWM)用于大鼠空间学习记忆。结果与对照组相比，HTO显著降低了神经细胞骨架蛋白F-actin (t=8.898 ~ 19.896, P<0.05)、α-微管蛋白(t=3.261 ~ 7.900, P<0.05)、tau (t=2.274 ~ 5.003, P<0.05)、MAP2 (t=2.274 ~ 5.003, P<0.05)和BDNF(t=3.580 ~ 19.792, P<0.05)、Reelin (t=3.240 ~ 39.692, P<0.05) mRNA的表达，且呈剂量依赖性。此外，MWM试验显著延长了照射子代的逃避潜伏期(t=-2.563, P<0.05)，显著减少了照射子代穿过目标象限的时间(t=3.214, P<0.05)，显著缩短了照射子代在平台象限的游泳时间(t=3.874, P<0.05)。结论子宫内HTO照射可下调神经元迁移相关因子的表达，引起脑功能障碍，这可能为辐射致脑损伤的机制提供线索。关键词:神经元细胞;氚化水;神经元迁移相关因素;空间学习与记忆

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