Brain-derived neurotrophic factor-tyrosine kinase B pathway mediates NMDA receptor NR2B subunit phosphorylation in the supraoptic nuclei following progressive dehydration.
Brain-derived neurotrophic factor-tyrosine kinase B pathway mediates NMDA receptor NR2B subunit phosphorylation in the supraoptic nuclei following progressive dehydration.
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DOI：
10.1111/j.1365-2826.2011.02209.x
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发表时间：
2011-10
影响因子：
3.2
通讯作者：
Cunningham JT
中科院分区：
文献类型：
--
作者：
Carreño FR;Walch JD;Dutta M;Nedungadi TP;Cunningham JT
本研究探讨了水剥夺对大鼠视上核（SON）TrkB和NMDA受体亚单位磷酸化的影响。采用激光捕获显微切割和qRT-PCR技术检测加压素SON神经元中BDNF和TrkB基因表达。免疫组化证实BDNF染色后叶加压素神经元，而磷酸化的TrkB染色增加水剥夺。含有SON的脑穿孔的Western印迹分析显示，与对照组相比，WD动物中TrkB（pTrkBY 515）的酪氨酸磷酸化、NR 1（pNR 1 S866或pNR 1）的丝氨酸磷酸化和NR 2B亚基（pNR 2BY 1472或pNR 2B）的酪氨酸磷酸化显著增加。接触水2小时将pTrkBY 515含量降低至对照水平，而不影响pNR 1或pNR 2B。需要4小时的再水化以将pNR 1和pNR 2B减少至对照。为了测试本研究中TrkB的磷酸化增加是否由BDNF介导，一组动物装备有与填充有载体或TrkB-Fc融合蛋白的微型渗透泵偶联的右侧SON套管，所述载体或TrkB-Fc融合蛋白防止BDNF与TrkB结合。在左SON对侧的套管，TrkB磷酸化显着增强后WD。对接受TrkB-Fc的右侧SON的单独分析显示，WD后TrkB受体磷酸化显着减弱。虽然WD后增加的pNR 1 S866不受TrkB-Fc局部输注的影响，但pNR 2BY 1472显著降低。免疫共沉淀显示，增加Fyn激酶和NR 2B和TrkB之间的物理相互作用在SON后水剥夺。因此，WD后SON中TrkB的激活可能通过NR 2B亚基的磷酸化影响细胞兴奋性。
We studied the effects of water deprivation on the phosphorylation of TrkB and NMDA receptor subunits in the supraoptic nucleus (SON) of the rat. Laser capture microdissection and qRT-PCR was used to demonstrate BDNF and TrkB gene expression in vasopressin SON neurones. Immunohistochemistry confirmed BDNF staining in vasopressin neurones, while staining for phosphorylated TrkB was increased following water deprivation. Western Blot analysis of brain punches containing the SON revealed that tyrosine phosphorylation of TrkB (pTrkBY515), serine phosphorylation of NR1 (pNR1S866 or pNR1) and tyrosine phosphorylation of NR2B subunits (pNR2BY1472 or pNR2B) were significantly increased in WD animals compared to control. Access to water for 2 h reduced pTrkBY515 content to control levels without affecting pNR1 or pNR2B. Four hours of rehydration was needed to reduce pNR1 and pNR2B to control. To test whether increased phosphorylation of TrkB in this study is mediated by BDNF, a group of animals were instrumented with right SON cannula coupled to mini-osmotic pumps filled with vehicle or TrkB-Fc fusion protein which prevents BDNF binding to TrkB. In the left SON contralateral to the cannula, TrkB phosphorylation was significantly enhanced following WD. Separate analysis of the right SON, which received TrkB-Fc, showed that the TrkB receptor phosphorylation following WD was significantly attenuated. While increased pNR1S866 following WD was not affected by local infusion of TrkB-Fc, pNR2BY1472 was significantly reduced. Co-immunoprecipitation revealed an increased physical interaction between Fyn kinase and NR2B and TrkB in the SON following water deprivation. Thus, activation of TrkB in the SON following WD may affect cellular excitability through the phosphorylation of NR2B subunits.
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DOI：
10.1016/s0169-328x(96)00205-7
发表时间：
1997-03-01
期刊：
MOLECULAR BRAIN RESEARCH
影响因子：
--
作者：
AlGhoul, WM;Meeker, RB;Greenwood, RS
通讯作者：
Greenwood, RS
影响因子：
5.5
作者：
Boudaba, C;Linn, DM;Tasker, JG
通讯作者：
Tasker, JG
DOI：
10.1016/s0165-3806(98)00060-1
发表时间：
1998-07-01
期刊：
DEVELOPMENTAL BRAIN RESEARCH
影响因子：
--
作者：
Currás, MC;Dao, J
通讯作者：
Dao, J
DOI：
10.1016/s0169-328x(99)00129-1
发表时间：
1999-07-05
期刊：
MOLECULAR BRAIN RESEARCH
影响因子：
--
作者：
Currás-Collazo, M;Dao, J
通讯作者：
Dao, J
影响因子：
15.9
作者：
DUNN, FL;BRENNAN, TJ;ROBERTSON, GL
通讯作者：
ROBERTSON, GL