McMurray K M J, Du X, Brownlee M, Palmer A A
Committee on Neurobiology, University of Chicago, Chicago, IL 60637, USA; Department of Human Genetics, University of Chicago, Chicago, IL 60637, USA.
Diabetes Research Center, Department of Medicine and Department of Pathology, Albert Einstein College of Medicine, Bronx, NY, USA.
Behav Brain Res. 2016 Mar 15;301:119-23. doi: 10.1016/j.bbr.2015.12.026. Epub 2015 Dec 19.
GLO1 (Glyoxalase1) is a ubiquitous cellular enzyme that detoxifies methylglyoxal (MG), which is a byproduct of glycolysis. Previously, we showed that ubiquitous overexpression of Glo1 reduced concentrations of MG and increased anxiety-like behavior, whereas systemic injection of MG reduced anxiety-like behavior. We further showed that MG is a competitive partial agonist at GABA-A receptors. Based on those data we hypothesized that modulation of GABAergic signaling by MG underlies Glo1 and MG's effects on anxiety-like behavior. As previous studies used ubiquitous overexpression, we sought to determine whether neuronal Glo1 overexpression was sufficient to increase anxiety-like behavior. We generated ROSA26 knock-in mice with a floxed-stop codon upstream from human Glo1 (FLOXGlo1KI) and bred them with mice expressing CRE recombinase under the direction of the Synapsin 1 promoter (Syn-CRE) to limit overexpression of Glo1 specifically to neurons. Furthermore, since previous administration of MG had been systemic, we sought to determine if direct microinjection of MG into the basolateral amygdala (BLA) was sufficient to reduce anxiety-like behavior. Thus, we performed bilateral microinjections of saline, MG (12μM or 24μM), or the positive control midazolam (4mM) directly into the BLA. FLOXGlo1KIxSyn-CRE mice showed significantly increased anxiety-like behavior compared to their FLOXGLO1xWT littermates. In addition, bilateral microinjection of MG and midazolam significantly decreased anxiety-like behavior compared to saline treated mice. These studies suggest that anatomically specific manipulations of Glo1 and MG are sufficient to induce changes in anxiety-like behavior.
乙二醛酶1(GLO1)是一种普遍存在于细胞中的酶,可对甲基乙二醛(MG)进行解毒,甲基乙二醛是糖酵解的副产物。此前,我们发现GLO1的普遍过表达会降低MG的浓度并增加焦虑样行为,而全身注射MG则会减少焦虑样行为。我们进一步表明,MG是GABA - A受体的竞争性部分激动剂。基于这些数据,我们推测MG对GABA能信号的调节是GLO1和MG对焦虑样行为产生影响的基础。由于先前的研究使用的是普遍过表达,我们试图确定神经元GLO1过表达是否足以增加焦虑样行为。我们构建了一种ROSA26基因敲入小鼠,其在人GLO1上游带有一个可诱导的终止密码子(FLOXGlo1KI),并将它们与在突触素1启动子(Syn - CRE)指导下表达CRE重组酶的小鼠进行杂交,以将GLO1的过表达特异性限制在神经元中。此外,由于之前给予MG是全身性的,我们试图确定直接将MG微量注射到基底外侧杏仁核(BLA)是否足以减少焦虑样行为。因此,我们将生理盐水、MG(12μM或24μM)或阳性对照咪达唑仑(4mM)双侧微量注射到BLA中。与它们的FLOXGLO1xWT同窝小鼠相比,FLOXGlo1KIxSyn - CRE小鼠表现出明显增加的焦虑样行为。此外,与生理盐水处理的小鼠相比,双侧微量注射MG和咪达唑仑显著降低了焦虑样行为。这些研究表明,对GLO1和MG进行解剖学上特定的操作足以引起焦虑样行为的改变。