慢性低压低氧暴露对小鼠海马CA1区神经元树突棘形态及细丝蛋白A表达的影响
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摘要
目的:探讨低压低氧暴露对小鼠海马CA1区神经元树突棘形态及细丝蛋白A表达的影响。方法:6～8周龄C57BL/6雄性小鼠分为常氧暴露7 d组、常氧暴露14 d组、低压低氧暴露7 d组和低压低氧暴露14 d组。低压低氧暴露组置于低压舱模拟6 000 m海拔高原进行低压低氧暴露。Golgi染色法观察小鼠海马CA1区树突的分支数,以及基树突棘和顶树突棘长度和密度的变化; Western blot方法检测小鼠海马细丝蛋白A表达水平的变化;免疫组织荧光染色法检测小鼠海马CA1区细丝蛋白A的表达及分布变化。结果:与常氧暴露组相比,低压低氧暴露后,小鼠海马CA1区树突分支数的差异无统计学显著性,但基树突棘和顶树突棘的长度显著增加(P <0. 05),密度显著降低(P <0. 01)。低压低氧暴露后,小鼠海马细丝蛋白A表达水平低于常氧暴露组(P <0. 01或P<0. 05)。免疫组织荧光染色显示细丝蛋白A在小鼠海马CA1区表达,低压低氧暴露后,海马CA1区细丝蛋白A表达水平降低(P <0. 05)。结论:慢性低压低氧暴露可影响小鼠海马CA1区细丝蛋白A表达,并导致海马CA1区神经元树突棘形态发生改变。
AIM: To investigate the effects of hypobaric hypoxic exposure on the morphological changes of dendritic spines and the expression of filamin-A in the neurons of mouse hippocampal CA1 region. METHODS: C57 BL/6 male mice( 6 ～ 8-week-old) were divided into normoxia 7 d group,normoxia 14 d group,hypobaric hypoxia 7 d group and hypobaric hypoxia 14 d group. The mice in hypobaric hypoxia exposure groups were placed in a hypobaric chamber with hypobaric hypoxia exposure to simulate the plateau at an altitude of 6 000 m. Golgi staining assay was used to observe the branch number of dendrites,and the length and density of basal and apical dendritic spines in the hippocampal CA1 region. The protein expression of filamin-A in the hippocampus of the mice was determined by Western blot. The protein expression and distribution of filamin-A in the hippocampal CA1 region were detected by immunofluorescence staining. RESULTS: Compared with normoxia exposure group,no significant difference of the number of dendritic branches in the hippocampal CA1 region after hypobaric hypoxia exposure was observed. However,the length of basal spines and apical spines was increased significantly( P < 0. 05),and the density of basal spines and apical spines was significantly reduced after hypobaric hypoxia exposure( P < 0. 01). The results of Western blot showed that the protein expression of filamin-A in the hippocampus of the mice after hypobaric hypoxia exposure was lower than that in normoxia exposure group( P < 0. 01 or P < 0. 05). Immunofluorescence staining showed that the filamin-A protein was expressed in the mouse hippocampal CA1 region,and the expression level after hypobaric hypoxia exposure was lower than that in normoxia group. CONCLUSION:Chronic hypobaric hypoxia exposure affects the protein expression level of filamin-A in the mouse hippocampal CA1 region,thus leading to the morphological changes of dendritic spines in the hippocampal CA1 region.
AIM: To investigate the effects of hypobaric hypoxic exposure on the morphological changes of dendritic spines and the expression of filamin-A in the neurons of mouse hippocampal CA1 region. METHODS: C57 BL/6 male mice( 6 ～ 8-week-old) were divided into normoxia 7 d group,normoxia 14 d group,hypobaric hypoxia 7 d group and hypobaric hypoxia 14 d group. The mice in hypobaric hypoxia exposure groups were placed in a hypobaric chamber with hypobaric hypoxia exposure to simulate the plateau at an altitude of 6 000 m. Golgi staining assay was used to observe the branch number of dendrites,and the length and density of basal and apical dendritic spines in the hippocampal CA1 region. The protein expression of filamin-A in the hippocampus of the mice was determined by Western blot. The protein expression and distribution of filamin-A in the hippocampal CA1 region were detected by immunofluorescence staining. RESULTS: Compared with normoxia exposure group,no significant difference of the number of dendritic branches in the hippocampal CA1 region after hypobaric hypoxia exposure was observed. However,the length of basal spines and apical spines was increased significantly( P < 0. 05),and the density of basal spines and apical spines was significantly reduced after hypobaric hypoxia exposure( P < 0. 01). The results of Western blot showed that the protein expression of filamin-A in the hippocampus of the mice after hypobaric hypoxia exposure was lower than that in normoxia exposure group( P < 0. 01 or P < 0. 05). Immunofluorescence staining showed that the filamin-A protein was expressed in the mouse hippocampal CA1 region,and the expression level after hypobaric hypoxia exposure was lower than that in normoxia group. CONCLUSION:Chronic hypobaric hypoxia exposure affects the protein expression level of filamin-A in the mouse hippocampal CA1 region,thus leading to the morphological changes of dendritic spines in the hippocampal CA1 region.
引文
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[14]黄辉,阮怀珍,范晓棠,等.低压低氧对胎鼠海马神经元NMDA受体影响的实验研究[J].中国应用生理学杂志,2002,18(4):321-323.
[15] Kandikattu HK,Deep SN,Razack S,et al. Hypoxia induced cognitive impairment modulating activity of Cyperus rotundus[J]. Physiol Behav,2017,175:56-65.
[16]梁冬施,陈利亚,洪芳芳,等.慢性间歇性低氧对幼鼠脑区AMPK通路的影响[J].中国病理生理杂志,2016,32(7):1200-1207.
[17] Nimchinsky EA,Sabatini BL,Svoboda K,et al. Structure and function of dendritic spines[J]. Annu Rev Phisiol,2002,64:313-353.
[18]李颖,向瑶,邓惠萍,等.白藜芦醇对花萼海绵诱癌素所致小鼠空间记忆障碍的预防作用[J].中国病理生理杂志,2017,33(5):951-955.
[19] Segura I,Lange C,Knevels E,et al. The oxygen sensor PHD2 controls dendritic spines and synapses via modification of filamin A[J]. Cell Rep,2016,14(11):2653-2667.
[20] Yagi H,Nagano T,Xie MJ,et al. Filamin A-interacting protein(FILIP)is a region-specific modulator of myosin2b and controls spine morphology and NMDA receptor accumulation[J]. Sci Rep,2014,4:6353.
[21] Zhang L,Bartley CM,Gong X,et al. MEK-ERK1/2-dependent FLNA overexpression promotes abnormal dendritic patterning in tuberous sclerosis independent of m TOR[J].Neuron,2014,84(1):78-91.
[22] Zheng L,Michelson Y,Freger V,et al. Drosophila Tenm and filamin affect motor neuron growth cone guidance[J]. PLo S One,2011,6(8):e22956.
[23] Sarkisian MR,Bartley CM,Chi H,et al. MEKK4 signaling regulates filamin expression and neuronal migration[J]. Neuron,2006,52(5):789-801.
[2]黎博,魏红艳,杨焰,等.神经干细胞外泌体对低氧所致神经元凋亡的抑制作用[J].中国病理生理杂志,2018,34(4):717-722.
[3] Bogdanova OV,Abdullah O,Kanekar S,et al. Neurochemical alterations in frontal cortex of the rat after one week of hypobaric hypoxia[J]. Behav Brain Res,2014,263:203-209.
[4] Qaid E,Zakaria R,Sulaiman SF. Insight into potential mechanisms of hypobaric hypoxia-induced learning and memory deficit:lessons from rat studies[J]. Hum Exp Toxicol,2017,36(12):1315-1325.
[5] Holtmaat A,Svoboda K. Experience-dependent structural synaptic plasticity in the mammalian brain[J]. Nat Rev Neurosci,2009,10(9):647-658.
[6] Hota SK,Barhwal K,Singh SB,et al. NR1 and GluR2expression mediates excitotoxicity in chronic hypobaric hypoxia[J]. J Neurosci Res,2008,86(5):1142-1152.
[7] Gipson CD,Olive MF. Structural and functional plasticity of dendritic spines:root or result of behavior[J]. Genes Brain Behav,2017,16(1):101-117.
[8] Nakamura F,Stossel TP,Hartwig JH,et al. The filamins:organizers of cell structure and function[J]. Cell Adh Migr,2011,5(2):160-169.
[9]朱丹,项世龙,薛智敏,等.细丝蛋白A在细胞黏附和迁移中的作用[J].中国细胞生物学学报,2014,36(8):1142-1152.
[10]董小铷,张向楠,李丹,等.红景天苷对低压低氧诱发大鼠脑损伤的保护作用[J].细胞与分子免疫学杂志,2015,31(10):1327-1331.
[11]王殿仕,吕顺艳,洪桢.大鼠海马CA1区锥体神经元树突分支的生后发育[J].解剖学报,2005,26(1):28-31.
[12]李慧,黄景阳,袁中瑞.提高脑组织冰冻切片质量的几点体会[J].中国免疫学杂志,2012,28(11):1019-1022.
[13] Czerniczyniec A,La Padula P,Bustamante J,et al. Mitochondrial function in rat cerebral cortex and hippocampus after short-and long-term hypobaric hypoxia[J]. Brain Res,2015,1598:66-75.
[14]黄辉,阮怀珍,范晓棠,等.低压低氧对胎鼠海马神经元NMDA受体影响的实验研究[J].中国应用生理学杂志,2002,18(4):321-323.
[15] Kandikattu HK,Deep SN,Razack S,et al. Hypoxia induced cognitive impairment modulating activity of Cyperus rotundus[J]. Physiol Behav,2017,175:56-65.
[16]梁冬施,陈利亚,洪芳芳,等.慢性间歇性低氧对幼鼠脑区AMPK通路的影响[J].中国病理生理杂志,2016,32(7):1200-1207.
[17] Nimchinsky EA,Sabatini BL,Svoboda K,et al. Structure and function of dendritic spines[J]. Annu Rev Phisiol,2002,64:313-353.
[18]李颖,向瑶,邓惠萍,等.白藜芦醇对花萼海绵诱癌素所致小鼠空间记忆障碍的预防作用[J].中国病理生理杂志,2017,33(5):951-955.
[19] Segura I,Lange C,Knevels E,et al. The oxygen sensor PHD2 controls dendritic spines and synapses via modification of filamin A[J]. Cell Rep,2016,14(11):2653-2667.
[20] Yagi H,Nagano T,Xie MJ,et al. Filamin A-interacting protein(FILIP)is a region-specific modulator of myosin2b and controls spine morphology and NMDA receptor accumulation[J]. Sci Rep,2014,4:6353.
[21] Zhang L,Bartley CM,Gong X,et al. MEK-ERK1/2-dependent FLNA overexpression promotes abnormal dendritic patterning in tuberous sclerosis independent of m TOR[J].Neuron,2014,84(1):78-91.
[22] Zheng L,Michelson Y,Freger V,et al. Drosophila Tenm and filamin affect motor neuron growth cone guidance[J]. PLo S One,2011,6(8):e22956.
[23] Sarkisian MR,Bartley CM,Chi H,et al. MEKK4 signaling regulates filamin expression and neuronal migration[J]. Neuron,2006,52(5):789-801.