Leukocyte trafficking through the choroid plexus as modulator of neonatal focal stroke

摘要 围产期动脉缺血性卒中是一种常见的疾病，会导致严重的发病率和严重的长期并发症 神经缺陷。围产期中风导致显著的发病率和严重的长期神经疾病 和认知缺陷。超过一半的脑性瘫痪儿童是在足月出生的。它已经变成了 明确损伤时大脑的成熟阶段在大脑模式中起着关键作用 围产期卒中不同病理生理学的潜在机制 成人中的中风仍然知之甚少。脉络膜神经丛(CPs)对大脑的保护作用 物理和生化障碍，并通过改善神经炎症。直到最近，CP还在 被忽视的潜在治疗靶点，但现在被认为是 神经退行性疾病的治疗。我们将研究CPS的神经保护作用 用一过性大脑中动脉建立适合年龄的卒中模型预防新生儿卒中 闭塞(TMCAO)在出生后第9天(P9)小鼠。特别是，我们将确定子集如何 单核细胞通过CPS的保护作用是否与Toll样受体2(TLR2)有关 CPS是先天免疫系统的组成部分，在CPS介导的神经保护中起着核心作用。 我们假设单核细胞通过CPS转运保护新生儿大脑免受tMCAO的侵袭。 以依赖于TLR2的方式。 我们将确定通过CPS进入的单核细胞表型与 新生鼠TMCAO后炎症和损伤(Aim1)及其机制的研究 损伤时通过功能或功能障碍的CPS介导的单核细胞转运(目标2)。我们将使用Unique Cx3Cr1GFP/+/CCR2RF/P+小鼠对单核细胞和小胶质细胞的区分及鉴定 单核细胞/小胶质细胞表型及利用双报告基因LUC/GFP-TLR2小鼠进行无创性监测 活体损伤小鼠TLR2实时上调的时空模式。我们有能力 磁共振弥散加权成像(DWI)结合多种成像技术识别新生鼠TMCAO损伤 多色流式细胞术、EMSA和细胞因子复合检测等生化检测 抑制CPS或/和TLR2的功能将增强对CPS作为保护作用的理解 新生儿卒中的屏障，为如何减轻新生儿急性卒中后的损伤提供新的见解 并确定血液-脑脊液屏障的药理靶点。

Abstract Perinatal arterial ischemic stroke is common and produces significant morbidity and severe long-term neurological deficits. Perinatal stroke leads to significant morbidity and severe long-term neurological and cognitive deficits. More than half of all children with cerebral palsy are born at term. It has become clear that the maturational stage of the brain at the time of injury plays a key role in the pattern of brain damage in humans but the underlying mechanisms of the differing pathophysiology of perinatal stroke and stroke in the adult are still poorly understood. The choroid plexuses (CPs) may protect the brain as physical and biochemical barriers and by modifying neuroinflammation. Until recently CPs were neglected as a potential target for therapy but are now considered as pharmacological target for treatment of neurodegenerative diseases. We will investigate the neuroprotective role of the CPs against neonatal stroke induced in an age-appropriate stroke model, a transient middle cerebral artery occlusion (tMCAO) in postnatal day 9 (P9) mice. In particular, we will determine how subsets of monocytes that enter through the CPs protect and whether toll-like receptor 2 (TLR2), one of key components of the innate immune system, plays a central role in CPs-mediated neuroprotection. We hypothesize that monocyte trafficking through CPs protects neonatal brain against tMCAO in a TLR2 dependent manner. We will determine relationships between the monocyte phenotypes entering through CPs and inflammation and injury after tMCAO in neonatal mice (Aim1) and define the mechanisms of TLR2- mediated monocyte trafficking via functional or dysfunctional CPs on injury (Aim 2). We will use unique Cx3Cr1GFP/+/CCR2RF/P+ mice to distinguish monocytes from microglia and characterize monocyte/microglial phenotypes and use double-reporter luc/GFP-TLR2 mice to non-invasively monitor the spatial-temporal pattern of TLR2 upregulation in real time in living injured mice. Our ability to identify injured neonatal mice during tMCAO by diffusion-weighted MRI (DWI) combined with various biochemical assays, such as multi-color flow cytometry, EMSA and cytokine multiplex, in mice with inhibited function of CPs or/and TLR2 will enhance the understanding of the role of CPs as protective barrier in neonatal stroke and provide new insights on how to alleviate injury after acute neonate stroke and identify pharmacological targets at the blood-CSF barrier.