Aix Marseille Univ, INSERM, INMED, 13273 Marseille, France.
Neuroscience Center, 00014 University of Helsinki, Helsinki, Finland.
Brain. 2023 Oct 3;146(10):4247-4261. doi: 10.1093/brain/awad132.
Although the Na-K-Cl cotransporter (NKCC1) inhibitor bumetanide has prominent positive effects on the pathophysiology of many neurological disorders, the mechanism of action is obscure. Attention paid to elucidating the role of Nkcc1 has mainly been focused on neurons, but recent single cell mRNA sequencing analysis has demonstrated that the major cellular populations expressing NKCC1 in the cortex are non-neuronal. We used a combination of conditional transgenic animals, in vivo electrophysiology, two-photon imaging, cognitive behavioural tests and flow cytometry to investigate the role of Nkcc1 inhibition by bumetanide in a mouse model of controlled cortical impact (CCI). Here, we found that bumetanide rescues parvalbumin-positive interneurons by increasing interneuron-microglia contacts shortly after injury. The longitudinal phenotypic changes in microglia were significantly modified by bumetanide, including an increase in the expression of microglial-derived BDNF. These effects were accompanied by the prevention of CCI-induced decrease in hippocampal neurogenesis. Treatment with bumetanide during the first week post-CCI resulted in significant recovery of working and episodic memory as well as changes in theta band oscillations 1 month later. These results disclose a novel mechanism for the neuroprotective action of bumetanide mediated by an acceleration of microglial activation dynamics that leads to an increase in parvalbumin interneuron survival following CCI, possibly resulting from increased microglial BDNF expression and contact with interneurons. Salvage of interneurons may normalize ambient GABA, resulting in the preservation of adult neurogenesis processes as well as contributing to bumetanide-mediated improvement of cognitive performance.
尽管钠-钾-2 氯共转运蛋白(NKCC1)抑制剂布美他尼对许多神经疾病的病理生理学有显著的积极影响,但作用机制尚不清楚。人们对 Nkcc1 作用的关注主要集中在神经元上,但最近的单细胞 mRNA 测序分析表明,皮质中表达 NKCC1 的主要细胞群体是非神经元。我们使用条件性转基因动物、体内电生理学、双光子成像、认知行为测试和流式细胞术,研究布美他尼抑制 NKCC1 在皮质控制冲击(CCI)小鼠模型中的作用。在这里,我们发现布美他尼通过增加损伤后短时间内神经元-小胶质细胞的接触来挽救 PV 阳性中间神经元。布美他尼显著改变了小胶质细胞的纵向表型变化,包括小胶质细胞衍生的 BDNF 的表达增加。这些效应伴随着 CCI 诱导的海马神经发生减少的预防。CCI 后第一周内用布美他尼治疗可显著恢复工作记忆和情景记忆,以及 1 个月后θ带振荡的变化。这些结果揭示了布美他尼神经保护作用的新机制,这种作用是通过加速小胶质细胞激活动力学介导的,导致 CCI 后 PV 阳性中间神经元的存活增加,可能是由于小胶质细胞 BDNF 表达和与中间神经元的接触增加所致。中间神经元的挽救可能使周围 GABA 正常化,从而维持成年神经发生过程,并有助于布美他尼介导的认知表现改善。