Suryavanshi Pratyush S, Gupta Subhash C, Yadav Roopali, Kesherwani Varun, Liu Jinxu, Dravid Shashank M
Department of Pharmacology, Creighton University, Omaha, Nebraska.
Department of Pharmacology, Creighton University, Omaha, Nebraska
Mol Pharmacol. 2016 Aug;90(2):96-105. doi: 10.1124/mol.116.104786. Epub 2016 May 26.
The delta family of ionotropic glutamate receptors consists of glutamate delta-1 (GluD1) and glutamate delta-2 receptors. We have previously shown that GluD1 knockout mice exhibit features of developmental delay, including impaired spine pruning and switch in the N-methyl-D-aspartate receptor subunit, which are relevant to autism and other neurodevelopmental disorders. Here, we identified a novel role of GluD1 in regulating metabotropic glutamate receptor 5 (mGlu5) signaling in the hippocampus. Immunohistochemical analysis demonstrated colocalization of mGlu5 with GluD1 punctas in the hippocampus. Additionally, GluD1 protein coimmunoprecipitated with mGlu5 in the hippocampal membrane fraction, as well as when overexpressed in human embryonic kidney 293 cells, demonstrating that GluD1 and mGlu5 may cooperate in a signaling complex. The interaction of mGlu5 with scaffold protein effector Homer, which regulates mechanistic target of rapamycin (mTOR) signaling, was abnormal both under basal conditions and in response to mGlu1/5 agonist (RS)-3,5-dihydroxyphenylglycine (DHPG) in GluD1 knockout mice. The basal levels of phosphorylated mTOR and protein kinase B, the signaling proteins downstream of mGlu5 activation, were higher in GluD1 knockout mice, and no further increase was induced by DHPG. We also observed higher basal protein translation and an absence of DHPG-induced increase in GluD1 knockout mice. In accordance with a role of mGlu5-mediated mTOR signaling in synaptic plasticity, DHPG-induced internalization of surface α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor subunits was impaired in the GluD1 knockout mice. These results demonstrate that GluD1 interacts with mGlu5, and loss of GluD1 impairs normal mGlu5 signaling potentially by dysregulating coupling to its effector. These studies identify a novel role of the enigmatic GluD1 subunit in hippocampal function.
离子型谷氨酸受体的δ家族由谷氨酸δ-1(GluD1)和谷氨酸δ-2受体组成。我们之前已经表明,GluD1基因敲除小鼠表现出发育延迟的特征,包括脊柱修剪受损和N-甲基-D-天冬氨酸受体亚基的转换,这些都与自闭症和其他神经发育障碍相关。在此,我们确定了GluD1在调节海马体中代谢型谷氨酸受体5(mGlu5)信号传导方面的新作用。免疫组织化学分析表明,mGlu5与海马体中GluD1点状结构共定位。此外,GluD1蛋白在海马体膜组分中与mGlu5进行了共免疫沉淀,并且在人胚肾293细胞中过表达时也是如此,这表明GluD1和mGlu5可能在信号复合物中协同作用。在基础条件下以及对GluD1基因敲除小鼠给予mGlu1/5激动剂(RS)-3,5-二羟基苯甘氨酸(DHPG)时,mGlu5与调节雷帕霉素作用靶点(mTOR)信号传导的支架蛋白效应分子荷马的相互作用均异常。mGlu5激活下游的信号蛋白磷酸化mTOR和蛋白激酶B的基础水平在GluD1基因敲除小鼠中较高,并且DHPG未诱导其进一步增加。我们还观察到GluD1基因敲除小鼠的基础蛋白翻译较高,且DHPG未诱导其增加。与mGlu5介导的mTOR信号传导在突触可塑性中的作用一致,在GluD1基因敲除小鼠中,DHPG诱导的表面α-氨基-3-羟基-5-甲基-4-异恶唑丙酸受体亚基内化受损。这些结果表明,GluD1与mGlu5相互作用,并且GluD1的缺失可能通过失调与其效应分子的偶联而损害正常的mGlu5信号传导。这些研究确定了神秘的GluD1亚基在海马体功能中的新作用。