Acute neuroinflammation induces AIS structural plasticity in a NOX2-dependent manner

Background Chronic microglia-mediated inflammation and oxidative stress are well-characterized underlying factors in neurodegenerative disease, whereby reactive inflammatory microglia enhance ROS production and impact neuronal integrity. Recently, it has been shown that during chronic inflammation, neuronal integrity is compromised through targeted disruption of the axon initial segment (AIS), the axonal domain critical for action potential initiation. AIS disruption was associated with contact by reactive inflammatory microglia which wrap around the AIS, increasing association with disease progression. While it is clear that chronic microglial inflammation and enhanced ROS production impact neuronal integrity, little is known about how acute microglial inflammation influences AIS stability. Here, we demonstrate that acute neuroinflammation induces AIS structural plasticity in a ROS-mediated and calpain-dependent manner. Methods C57BL/6J and NOX2−/− mice were given a single injection of lipopolysaccharide (LPS; 5 mg/kg) or vehicle (0.9% saline, 10 mL/kg) and analyzed at 6 h–2 weeks post-injection. Anti-inflammatory Didox (250 mg/kg) or vehicle (0.9% saline, 10 mL/kg) was administered beginning 24 h post-LPS injection and continued for 5 days; animals were analyzed 1 week post-injection. Microglial inflammation was assessed using immunohistochemistry (IHC) and RT-qPCR, and AIS integrity was quantitatively analyzed using ankyrinG immunolabeling. Data were statistically compared by one-way or two-way ANOVA where mean differences were significant as assessed using Tukey’s post hoc analysis. Results LPS-induced neuroinflammation, characterized by enhanced microglial inflammation and increased expression of ROS-producing enzymes, altered AIS protein clustering. Importantly, inflammation-induced AIS changes were reversed following resolution of microglial inflammation. Modulation of the inflammatory response using anti-inflammatory Didox, even after significant AIS disruption occurred, increased the rate of AIS recovery. qPCR and IHC analysis revealed that expression of microglial NOX2, a ROS-producing enzyme, was significantly increased correlating with AIS disruption. Furthermore, ablation of NOX2 prevented inflammation-induced AIS plasticity, suggesting that ROS drive AIS structural plasticity. Conclusions In the presence of acute microglial inflammation, the AIS undergoes an adaptive change that is capable of spontaneous recovery. Moreover, recovery can be therapeutically accelerated. Together, these findings underscore the dynamic capabilities of this domain in the presence of a pathological insult and provide evidence that the AIS is a viable therapeutic target

Acute neuroinflammation induces AIS structural plasticity in a NOX2-dependent manner

Abstract Background Chronic microglia-mediated inflammation and oxidative stress are well-characterized underlying factors in neurodegenerative disease, whereby reactive inflammatory microglia enhance ROS production and impact neuronal integrity. Recently, it has been shown that during chronic inflammation, neuronal integrity is compromised through targeted disruption of the axon initial segment (AIS), the axonal domain critical for action potential initiation. AIS disruption was associated with contact by reactive inflammatory microglia which wrap around the AIS, increasing association with disease progression. While it is clear that chronic microglial inflammation and enhanced ROS production impact neuronal integrity, little is known about how acute microglial inflammation influences AIS stability. Here, we demonstrate that acute neuroinflammation induces AIS structural plasticity in a ROS-mediated and calpain-dependent manner. Methods C57BL/6J and NOX2−/− mice were given a single injection of lipopolysaccharide (LPS; 5 mg/kg) or vehicle (0.9% saline, 10 mL/kg) and analyzed at 6 h–2 weeks post-injection. Anti-inflammatory Didox (250 mg/kg) or vehicle (0.9% saline, 10 mL/kg) was administered beginning 24 h post-LPS injection and continued for 5 days; animals were analyzed 1 week post-injection. Microglial inflammation was assessed using immunohistochemistry (IHC) and RT-qPCR, and AIS integrity was quantitatively analyzed using ankyrinG immunolabeling. Data were statistically compared by one-way or two-way ANOVA where mean differences were significant as assessed using Tukey’s post hoc analysis. Results LPS-induced neuroinflammation, characterized by enhanced microglial inflammation and increased expression of ROS-producing enzymes, altered AIS protein clustering. Importantly, inflammation-induced AIS changes were reversed following resolution of microglial inflammation. Modulation of the inflammatory response using anti-inflammatory Didox, even after significant AIS disruption occurred, increased the rate of AIS recovery. qPCR and IHC analysis revealed that expression of microglial NOX2, a ROS-producing enzyme, was significantly increased correlating with AIS disruption. Furthermore, ablation of NOX2 prevented inflammation-induced AIS plasticity, suggesting that ROS drive AIS structural plasticity. Conclusions In the presence of acute microglial inflammation, the AIS undergoes an adaptive change that is capable of spontaneous recovery. Moreover, recovery can be therapeutically accelerated. Together, these findings underscore the dynamic capabilities of this domain in the presence of a pathological insult and provide evidence that the AIS is a viable therapeutic target

Mutual replacement reactions in alkali feldspars II: trace element partitioning and geothermometry

Perthitic alkali feldspar primocrysts in layered syenites in the Klokken intrusion in South Greenland, underwent dissolution–reprecipitation reactions in a circulating post-magmatic aqueous fluid at ∼450°C, and are to a large degree pseudomorphs. These 'mutual replacement' reactions provide a perfect natural experiment with which to study trace element partitioning between sodium and potassium feldspars growing simultaneously. The reactant 'phase' was a cryptoperthitic feldspar consisting of low albite and low microcline in a coherent sub-μm 'braid' intergrowth and the product phases were 'strain-free' incoherent subgrains of low albite and low microcline forming microporous patch perthites on scales up to 200 μm. The driving force for the reaction was reduction of coherency strain energy. The mechanisms of this process are described in Part I. Five mixed braid perthite–patch perthite crystals were analysed for major and trace elements using laser ablation-inductively coupled plasma mass spectrometry with a 19 μm beam diameter. This gave bulk analyses of the braid texture, which were in the range Ab73–54Or45–27An4.3–0.8, but could resolve Ab- and Or-rich patches in patch perthite. The major element bulk compositions of the crystals were retained during the replacement reactions. Major components in patches plot on tielines in the Ab–Or–An ternary system that pass through or very close to the parent braid perthite composition and indicate local equilibrium on the scale of a few tens of mm. Many trace elements, including REE, were lost to the fluid during the deuteric reactions, but the effect is large only for Fe and Ti. Cs, Pb and Sr were added to some crystals. Plots of log distribution coefficient D for Rb, Ba, Pb, Eu2+, La and Ce between Or- and Ab-rich patches against ionic radius are straight lines, assuming eightfold coordination, and to a first approximation are independent of ionic charge. K also lies on these lines, and the smaller ions Na and Ca lie close to them. The best linear fits were obtained using ionic radii for [8]K and [8]Ca, but there is ambiguity as to whether [7]Na or [5]Na is most appropriate. The linear relationship shows that the listed trace elements are in the feldspar M-site rather than in inclusions. Tl is in M although an exact D could not be obtained. The very large Cs ion partitions strongly into the Or-rich phase but its D value appears to be less than predicted by extrapolation. The near-linearity arises because partitioning is occurring between two solids into sites which have similar Young's moduli, so that the parabolas that normally represent trace element partitioning between crystals and liquids (which have negligible shear strength) approximately cancel out. Ga and Be are in T-sites, as well as some of the Fe and Ti present, although part is in oxide inclusions. The site of Sc is unclear, but if structural it is likely to be T. Partitioning on M-sites is a potential geothermometer but because the effective size of the irregular M-site is defined by its K and (Na + Ca) contents, which are controlled by ternary solvus relationships, its calibration is not independent of conventional two-feldspar geothermometers. Trace elements may however provide a useful means of confirming that feldspar pairs are in equilibrium, and of recognising feldspar intergrowths produced by non-isochemical replacement rather than exsolution. Two-feldspar geothermometry for the ternary phases in the low-albite microcline patch perthites gives temperatures above the stability range of microcline, markedly so if a correction is made for Si–Al ordering. This is probably because current geothermometers are too sensitive to low concentrations of An in ordered Or-rich feldspars. This interpretation is supported by two-feldspar assemblages growing at known temperatures in geothermal systems and sedimentary basins