新生儿程序性疼痛破坏脊髓中KCC2的磷酸化

IF 2.3 4区 医学 Q2 DEVELOPMENTAL BIOLOGY
Mathilde Baudat, Elbert A. J Joosten, Sinno H. P. Simons, Daniël L. A. van den Hove, Renzo J. M. Riemens
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引用次数: 0

摘要

新生儿重症监护病房的新生儿程序性疼痛可导致中枢神经系统,特别是脊髓伤害感受网络的长期重塑。临床前研究表明,由于γ-氨基丁酸(GABA)能神经传递减少,脊髓中的抑制性与兴奋性平衡被破坏。在新生儿发育过程中,gaba能发生转移,这是由氯化钾共转运蛋白2 (KCC2)及其催产素受体(OXTR)依赖性丝氨酸940残基磷酸化(pKCC2)调控的。由于Oxtr的DNA甲基化对早期生活逆境(如新生儿程序性疼痛)很敏感,我们假设新生儿程序性疼痛会降低腰椎Oxtr甲基化,从而阻止KCC2和pKCC2的发育增加。采用重复性新生儿程序性疼痛大鼠模型,从出生后第0天(P)至第7天(P),每天4针扎左后爪。在P0和P10收集脊髓样本,通过Western blot分析KCC2和pKCC2的水平。此外,使用亚硫酸氢盐焦磷酸测序对脊柱Oxtr甲基化进行量化。结果表明,新生儿程序性疼痛下调脊柱pKCC2水平,而KCC2水平保持不变。这些发现表明kcc2依赖性氯离子流出中断,并支持新生儿程序性疼痛破坏gaba能转移的假设。在P10动物的同侧脊髓中也观察到pKCC2/KCC2水平的发育降低,这表明在脊髓KCC2的发育调节中参与了其他翻译后机制。考虑到没有检测到Oxtr启动子甲基化的显著变化,Oxtr的甲基化似乎与gaba能转移的干扰无关。总体而言,本研究表明新生儿程序性疼痛破坏脊髓KCC2磷酸化,并支持新生儿程序性疼痛改变脊髓gaba能转移的假设。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Neonatal Procedural Pain Disrupts Phosphorylation of KCC2 in the Spinal Cord

Neonatal Procedural Pain Disrupts Phosphorylation of KCC2 in the Spinal Cord

Neonatal procedural pain experienced in the neonatal intensive care unit can lead to long-lasting remodeling of the central nervous system and, in particular, of the spinal nociceptive network. Preclinical studies indicate a disrupted inhibitory versus excitatory balance in the spinal cord due to reduced γ-aminobutyric acid (GABA) ergic neurotransmission. During neonatal development a GABAergic shift occurs, which is regulated by the potassium-chloride co-transporter 2 (KCC2) and its oxytocin receptor (OXTR)-dependent phosphorylation at the serine 940 residue (pKCC2). As DNA methylation of Oxtr is sensitive to early life adversity, such as neonatal procedural pain, we hypothesized that neonatal procedural pain reduces Oxtr methylation in the lumbar spinal cord and subsequently prevents the developmental increase in KCC2 and pKCC2. Using a rat model of repetitive neonatal procedural pain, four needle pricks were applied to the left hind paw every day from postnatal day (P)0 to P7. Spinal cord samples were collected at P0 and P10 to assess the levels of KCC2 and pKCC2 via Western blot analysis. Additionally, spinal Oxtr methylation was quantified using bisulfite pyrosequencing. The results indicated that neonatal procedural pain downregulates spinal pKCC2 levels, while KCC2 levels remain unchanged. These findings suggest a disrupted KCC2-dependent chloride outflow and support the hypothesis that neonatal procedural pain disrupts the GABAergic shift. A developmental decrease in pKCC2/KCC2 levels was also observed in the ipsilateral spinal cord of P10 animals, indicating the involvement of other post-translational mechanisms in the developmental regulation of spinal KCC2. Methylation of the Oxtr does not seem to be related to the disturbed GABAergic shift, given that no significant changes in Oxtr promoter methylation were detected. Overall, this study demonstrates that neonatal procedural pain disrupts spinal KCC2 phosphorylation and supports the hypothesis that neonatal procedural pain alters the GABAergic shift in the spinal cord.

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来源期刊
Developmental Neurobiology
Developmental Neurobiology 生物-发育生物学
CiteScore
6.50
自引率
0.00%
发文量
45
审稿时长
4-8 weeks
期刊介绍: Developmental Neurobiology (previously the Journal of Neurobiology ) publishes original research articles on development, regeneration, repair and plasticity of the nervous system and on the ontogeny of behavior. High quality contributions in these areas are solicited, with an emphasis on experimental as opposed to purely descriptive work. The Journal also will consider manuscripts reporting novel approaches and techniques for the study of the development of the nervous system as well as occasional special issues on topics of significant current interest. We welcome suggestions on possible topics from our readers.
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