尿素通道蛋白B敲除小鼠脑组织水通道蛋白4表达改变与抑郁样行为的关系

袁德智; 侯良绢; 田宽; 胡玲; 李晋芳; 冉建华

doi:10.16098/j.issn.0529-1356.2020.03.003

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解剖学报 ›› 2020, Vol. 51 ›› Issue (3) : 326-331. DOI: 10.16098/j.issn.0529-1356.2020.03.003

神经生物学

尿素通道蛋白B敲除小鼠脑组织水通道蛋白4表达改变与抑郁样行为的关系

袁德智^1,2侯良绢³田宽²胡玲²李晋芳^1*冉建华^2*

作者信息 +

Relationship between depression-like behavior and expression of aquaporin 4 in brain of urea transporter B null mice

YUAN De-zhi^1,2 HOU Liang-juan³ TIAN Kuan² HU Ling² LI Jin-fang^1*RAN Jian-hua^2*

Author information +

文章历史 +

摘要

目的探讨尿素通道蛋白B（UT-B）敲除小鼠海马超微结构改变，以及水通道蛋白4（AQP4）的表达变化，探讨形态改变与小鼠抑郁样行为的关系。方法采用糖水偏好和强迫游泳实验检测野生型与UT-B敲除小鼠（各10只）的行为差异，透射电子显微电镜观察小鼠海马组织的超微结构改变，免疫组织化学观察脑组织AQP4的表达，免疫印迹技术检测AQP4表达水平。结果野生型小鼠与UT-B基因敲除小鼠糖水偏好指数分别为(84.67±4.85)%和(65.67±7.97)%（P<0.001），而强迫游泳实验不动时间分别为(128.56±11.24) s和(209.11±20.99) s（P<0.001）；两种行为学检测结果显示，UT-B敲除小鼠表现出抑郁样行为。电子显微镜结果显示，敲除小鼠出现神经元神经纤维肿胀和退行性改变，血管周围星形胶质细胞终足肿胀。免疫组织化学结果显示，敲除小鼠脑皮质、海马、丘脑等部位AQP4免疫阳性细胞数量明显减少，AQP4免疫阳性细胞主要分布于软脑膜、室管膜及脑血管周围，但脑血管周围免疫染色减弱。免疫印迹法检测提示，海马组织中AQP4表达水平下调27.1%（P<0.05）。结论 UT-B 基因敲除小鼠脑皮质和海马AQP4表达减少，可能通过影响神经发生和脑代谢引起小鼠的抑郁样行为。

Abstract

Objective To investigate the ultrastructural changes of hippocampus in urea transporter B (UT-B) null mice and the alterations of distribution and expression level of aquaporin 4（AQP4）in brain, and to discuss the relationship between AQP4 expression changes and depression-like behaviors in UT-B null mice. Methods Behavior differences of wild-type and UT-B null mice（10 in each group） were detected with sucrose preference and forced swimming test. The ultrastructural changes of hippocampus were observed by transmission electron microscopy (TEM). Immunohistochemistry and Western blotting were performed to detect the distribution and expression level of AQP4 in both genotypes. Results The sucrose preference index of wild-type mice and UT-B null mice were (84.67±1.62)% and (65.67±2.66)%, respectively (P<0.001). The immobility time of forced swimming was (209.1±7.00) seconds and (128.6±3.75) seconds respectively (P<0.001). The two behavioral test results showed that UT-B null mice exhibited depression-like behavior. TEM results displayed the abnormal neurons with swelling of myelinated and unmyelinated fibers and degenerative changes, and perivascular astrocyte endfeet swelling. Immunohistochemistry results showed AQP4-immunoreactive (IR) cells were significantly reduced in cortex, hippocampus and thalamus. AQP4-IR cells were distributed in the pia matter, ependymal and cerebrovascular, but the perivascular immunostaining decreased. Western blotting analysis showed that the expression level of AQP4 in hippocampus was down-regulated by 27.1% (P<0.05). Conclusion Reduced expression of AQP4 in the cerebral cortex and hippocampus of UT-B null mice might induce depressive behaviors by inference neurogenesis and cerebral metabolism.

导出引用

袁德智侯良绢田宽胡玲李晋芳冉建华. 尿素通道蛋白B敲除小鼠脑组织水通道蛋白4表达改变与抑郁样行为的关系[J]. 解剖学报. 2020, 51(3): 326-331 https://doi.org/10.16098/j.issn.0529-1356.2020.03.003

YUAN De-zhi HOU Liang-juan TIAN Kuan HU Ling LI Jin-fang RAN Jian-hua. Relationship between depression-like behavior and expression of aquaporin 4 in brain of urea transporter B null mice[J]. Acta Anatomica Sinica. 2020, 51(3): 326-331 https://doi.org/10.16098/j.issn.0529-1356.2020.03.003

中图分类号： R361+2

参考文献

［1］ Hamilton NB, Attwell D. Do astrocytes really exocytose neurotransmitters?［J］. Nat Rev Neurosci, 2010,11(4):227-238.

［2］ Martin JL, Magistretti PJ, Allaman Ⅰ. Regulation of neurotrophic factors and energy metabolism by antidepressants in astrocytes［J］. Curr Drug Targets, 2013,14(11):1308-1321.

［3］ Cui W, Mizukami H, Yanagisawa M, et al. Glial dysfunction in the mouse habenula causes depressive-like behaviors and sleep disturbance［J］. J Neurosci, 2014,34(49): 16273-16285.

［4］ Cui Y, Yang Y, Ni Z, et al. Astroglial Kir4.1 in the lateral habenula drives neuronal bursts in depression［J］. Nature, 2018,554(7692): 323-327.

［5］ Westermair AL, Munz M, Schaich A, et al. Association of genetic variation at AQP4 locus with vascular depression［J］. Biomolecules, 2018,8(4): 164.

［6］ Rajkowska G, Miguel-Hidalgo JJ. Glial pathology in major depressive disorder: an approach to investigate the coverage of blood vessels by astrocyte endfeet in human postmortem brain［J］. Methods Mol Biol, 2019,1938:247-254.

［7］ Xia M, Yang L, Sun G, et al. Mechanism of depression as a risk factor in the development of Alzheimer’s disease: the function of AQP4 and the glymphatic system［J］. Psychopharmacology (Berl), 2017,234(3): 365-379.

［8］ Szu JI, Binder DK. The role of astrocytic aquaporin-4 in synaptic plasticity and learning and memory［J］. Front Integr Neurosci,2016,10:8.

［9］ Wu X, Zhang JT, Li D, et al. Aquaporin-4 deficiency facilitates fear memory extinction in the hippocampus through excessive activation of extrasynaptic GluN2B-containing NMDA receptors［J］. Neuropharmacology, 2017,112(Pt A):124-134.

［10］ Medina A, Watson SJ, Bunney W Jr, et al. Evidence for alterations of the glial syncytial function in major depressive disorder［J］. J Psychiatr Res, 2016,72:15-21.

［11］ Li X, Ran J, Zhou H, et al. Mice lacking urea transporter UT-B display depression-like behavior［J］. J Mol Neurosci, 2012,46(2):362-372.

[12］ Levin EJ, Quick M, Zhou M. Crystal structure of a bacterial homologue of the kidney urea transporter［J］. Nature, 2009,462(7274):757-761.

［13］ Pizzagalli DA. Depression, stress, and anhedonia: toward a synthesis and integrated model［J］. Annu Rev Clin Psychol, 2014,10:393-423.

［14］ Porsolt RD, Le Pichon M, Jalfre M. Depression: a new animal model sensitive to antidepressant treatments［J］. Nature, 1977,266(5604):730-732.

［15］ Otte C, Gold SM, Penninx BW, et al. Major depressive disorder［J］. Nat Rev Dis Primers, 2016,2:16065.

［16］ Blier P. Neurobiology of depression and mechanism of action of depression treatments［J］. J Clin Psychiatry, 2016,77(3):e319.

［17］ Wang Zh, Huang YJ, Nai AT, et al. Fluoxetine improves the depression-like behavior in mice induced by chronic restraint stress by up-regulating the expression of bromodomain protein 4 in hippocampus［J］.Acta Anatomica Sinica,2019,50(1): 18-23. (in Chinese)

王贞,黄怡佳,乃爱桃,等.氟西汀通过上调海马内溴结构域蛋白4的表达改善慢性束缚应激所致小鼠的抑郁样行为［J］.解剖学报,2019,50(1):18-23.

［18］ Vadodaria KC, Ji Y, Skime M, et al. Serotonin-induced hyperactivity in SSRI-resistant major depressive disorder patient-derived neurons［J］. Mol Psychiatry, 2019,24(6):795-807.

［19］ Rajkowska G, Hughes J, Stockmeier CA, et al. Coverage of blood vessels by astrocytic endfeet is reduced in major depressive disorder［J］. Biol Psychiatry, 2013,73(7):613-621.

［20］ Di Benedetto B, Malik VA, Begum S, et al. Fluoxetine requires the endfeet protein aquaporin-4 to enhance plasticity of astrocyte processes［J］. Front Cell Neurosci, 2016,10:8.