Department of Molecular and Integrative Physiology, School of Molecular and Cellular Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Neuroscience Program, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Neuropharmacology. 2018 Aug;138:182-192. doi: 10.1016/j.neuropharm.2018.06.011. Epub 2018 Jun 8.
Chronic activity perturbations in neurons induce homeostatic plasticity through modulation of synaptic strength or other intrinsic properties to maintain the correct physiological range of excitability. Although similar plasticity can also occur at the population level, what molecular mechanisms are involved remain unclear. In the current study, we utilized a multielectrode array (MEA) recording system to evaluate homeostatic neural network activity of primary mouse cortical neuron cultures. We demonstrated that chronic elevation of neuronal activity through the inhibition of GABA(A) receptors elicits synchronization of neural network activity and homeostatic reduction of the amplitude of spontaneous neural network spikes. We subsequently showed that this phenomenon is mediated by the ubiquitination of tumor suppressor p53, which is triggered by murine double minute-2 (Mdm2). Using a mouse model of fragile X syndrome, in which fragile X mental retardation protein (FMRP) is absent (Fmr1 knockout), we found that Mdm2-p53 signaling, network synchronization, and the reduction of network spike amplitude upon chronic activity stimulation were all impaired. Pharmacologically inhibiting p53 with Pifithrin-α or genetically employing p53 heterozygous mice to enforce the inactivation of p53 in Fmr1 knockout cultures restored the synchronization of neural network activity after chronic activity stimulation and partially corrects the homeostatic reduction of neural network spike amplitude. Together, our findings reveal the roles of both Fmr1 and Mdm2-p53 signaling in the homeostatic regulation of neural network activity and provide insight into the deficits of excitability homeostasis seen when Fmr1 is compromised, such as occurs with fragile X syndrome.
神经元的慢性活动扰动通过调节突触强度或其他内在特性诱导稳态可塑性,以维持正确的兴奋性生理范围。尽管类似的可塑性也可能发生在群体水平,但涉及哪些分子机制仍不清楚。在本研究中,我们利用多电极阵列 (MEA) 记录系统评估原代小鼠皮质神经元培养物的稳态神经网络活动。我们证明,通过抑制 GABA(A) 受体来慢性升高神经元活动会引发神经网络活动的同步和自发性神经网络尖峰幅度的稳态降低。随后我们表明,这种现象是由肿瘤抑制蛋白 p53 的泛素化介导的,而 p53 的泛素化是由鼠双微体 2 (Mdm2) 触发的。使用脆性 X 综合征的小鼠模型,其中缺乏脆性 X 智力低下蛋白 (FMRP)(Fmr1 敲除),我们发现 Mdm2-p53 信号通路、网络同步以及慢性活动刺激后网络尖峰幅度的降低均受到损害。用 Pifithrin-α 抑制 p53 或通过遗传手段在 Fmr1 敲除培养物中使用 p53 杂合子小鼠强制使 p53 失活,可恢复慢性活动刺激后的神经网络活动同步,并部分纠正神经网络尖峰幅度的稳态降低。总之,我们的发现揭示了 Fmr1 和 Mdm2-p53 信号通路在神经网络活动稳态调节中的作用,并深入了解了 Fmr1 受损时兴奋性稳态的缺陷,如脆性 X 综合征。