Traumatic brain injury (TBI) is traditionally characterized by primary and secondary injury phases, both contributing to pathological and morphological changes. The mechanisms of damage and chronic consequences of TBI remain to be fully elucidated, but synaptic homeostasis disturbances and impaired energy metabolism are proposed to be a major contributor. It has been proposed that an increase of extracellular (eATP) adenosine triphosphate (ATP) in the area immediately surrounding impact may play a pivotal role in this sequence of events. After tissue injury, rupture of cell membranes allows release of intracellular ATP into the extracellular space, triggering a cascade of toxic events and inflammation. ATP is a ubiquitous messenger; however, simple and reliable techniques to measure its concentration have proven elusive. Here, we integrate a sensitive bioluminescent eATP sensor known as pmeLUC, with a controlled cortical impact mouse model to monitor eATP changes in a living animal after injury. Using the pmeLUC probe, a rapid increase of eATP is observed proximal to the point of impact within minutes of the injury. This event is significantly attenuated when animals are pretreated with an ATP hydrolyzing agent (apyrase) before surgery, confirming the contribution of eATP. This new eATP reporter could be useful for understanding the role of eATP in the pathogenesis in TBI and may identify a window of opportunity for therapeutic intervention.

中文翻译：

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创伤性脑损伤小鼠模型中细胞外三磷酸腺苷的体内检测

创伤性脑损伤 (TBI) 传统上以原发性和继发性损伤阶段为特征，两者都会导致病理和形态变化。TBI 的损伤和慢性后果的机制仍有待充分阐明，但突触稳态紊乱和能量代谢受损被认为是主要因素。有人提出，紧邻撞击区域的细胞外 (eATP) 三磷酸腺苷 (ATP) 的增加可能在这一系列事件中起关键作用。组织损伤后，细胞膜破裂允许细胞内 ATP 释放到细胞外空间，引发一连串的毒性事件和炎症。ATP是无处不在的信使；然而，测量其浓度的简单而可靠的技术已被证明是难以捉摸的。这里，我们将称为 pmeLUC 的敏感生物发光 eATP 传感器与受控皮质冲击小鼠模型集成，以监测受伤后活体动物的 eATP 变化。使用 pmeLUC 探针，在受伤后的几分钟内，在撞击点附近观察到 eATP 的快速增加。当动物在手术前用 ATP 水解剂 (apyrase) 预处理时，该事件显着减弱，证实了 eATP 的贡献。这种新的 eATP 报告基因可能有助于了解 eATP 在 TBI 发病机制中的作用，并可能确定治疗干预的机会窗口。在受伤后的几分钟内，在撞击点附近观察到 eATP 的快速增加。当动物在手术前用 ATP 水解剂 (apyrase) 预处理时，该事件显着减弱，证实了 eATP 的贡献。这种新的 eATP 报告基因可能有助于了解 eATP 在 TBI 发病机制中的作用，并可能确定治疗干预的机会窗口。在受伤后的几分钟内，在撞击点附近观察到 eATP 的快速增加。当动物在手术前用 ATP 水解剂 (apyrase) 预处理时，该事件显着减弱，证实了 eATP 的贡献。这种新的 eATP 报告基因可能有助于了解 eATP 在 TBI 发病机制中的作用，并可能确定治疗干预的机会窗口。