We adapt a mouse global ischemia model to permit rapid induction of ischemia and reperfusion in conjunction with two-photon imaging to monitor the initial ionic, structural, and functional implications of brief interruptions of blood flow (6 -8 min) in vivo. After only 2-3 min of global ischemia, a wide spread loss of mouse somatosensory cortex apical dendritic structure is initiated during the passage of a propagating wave (3.3 mm/min) of ischemic depolarization. Increases in intracellular calcium levels occurred during the wave of ischemic depolarization and were coincident with the loss of dendritic structure, but were not triggered by reperfusion. To assess the role of NMDA receptors, we locally applied the antagonist MK-801 [(ϩ)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate] at concentrations sufficient to fully block local NMDA agonist-evoked changes in intracellular calcium levels in vivo. Changes in dendritic structure and intracellular calcium levels were independent of NMDA receptor activation. Local application of the non-NMDA glutamate receptor antagonist CNQX also failed to block ischemic depolarization or rapid changes in dendrite structure. Within 3-5 min of reperfusion, damage ceased and restoration of synaptic structure occurred over 10 -60 min. In contrast to a reperfusion promoting damage, over this time scale, the majority of spines and dendrites regained their original structure during reperfusion. Intrinsic optical signal imaging of sensory evoked maps indicated that reversible alteration in dendritic structure during reperfusion was accompanied by restored functional maps. Our results identify glutamate receptor-independent ischemic depolarization as the major ionic event associated with disruption of synaptic structure during the first few minutes of ischemia in vivo.

Despite significant developments in optical imaging of superoxide anion (O2(•-)) as the preliminary reactive oxygen species, novel visualizing strategies that offer ultrahigh sensitivity are still imperative. This is mainly because intrinsic concentrations of O2(•-) are extremely low in living systems. Herein, we present the rational design and construction of a new polymer nanoprobe PCLA-O2(•-) for detecting O2(•-) based on chemiluminescence (CL) resonance energy transfer without an external excitation source. Structurally, PCLA-O2(•-) contains two moieties linked covalently, namely imidazopyrazinone that is capable of CL triggered by O2(•-) as the energy donor and conjugated polymers with light-amplifying property as the energy acceptor. Experiment results demonstrate that PCLA-O2(•-) exhibits ultrahigh sensitivity at the picomole level, dramatically prolonged luminescence time, specificity, and excellent biocompatibility. Without exogenous stimulation, this probe for the first time in situ visualizes O2(•-) level differences between normal and tumor tissues of mice. These exceptional features ensure that PCLA-O2(•-) as a self-luminescing probe is an alternative in vivo imaging approach for ultralow level O2(•-).

Viscosity, as a vital microenvironment parameter, is tightly associated with multitudinous cellular processes and diseases. Recently, precise visualization of viscosity has started to arouse more and more interest. However, owing to the complicated character, it is still a huge challenge to directly observe viscosity in living systems. In this regard, mounting fluorescence probes are being increasingly fabricated to map viscosity inside live cells and small animals. In this minireview, the viscosity‐sensitive small molecular fluorescent probes used in bioimaging are comprehensively summarized, mainly focusing on the last three years. Moreover, the current challenges and opportunities for the development of viscosity‐specific fluorescent probes will be discussed.

Hepatic ischemia–reperfusion injury (HIRI) is responsible for postoperative liver dysfunction and liver failure. Precise and rapid navigation of HIRI lesions is critical for early warning and timely development of pretreatment plans. Available methods for assaying liver injury fail to provide the exact location of lesions in real time intraoperatively. HIRI is intimately associated with oxidative stress which impairs lysosomal degradative function, leading to significant changes in lysosomal viscosity. Therefore, lysosomal viscosity is a potential biomarker for the precise targeting of HIRI. Hence, we developed a viscosity-activatable second near-infrared window fluorescent probe (NP-V) for the detection of lysosomal viscosity in hepatocytes and mice during HIRI. A reactive oxygen species–malondialdehyde–cathepsin B signaling pathway during HIRI was established. We further conducted high signal-to-background ratio NIR-II fluorescence imaging of HIRI mice. The contour and boundary of liver lesions were delineated, and as such the precise intraoperative resection of the lesion area was implemented. This research demonstrates the potential of NP-V as a dual-functional probe for the elucidation of HIRI pathogenesis and the direct navigation of HIRI lesions in clinical applications.

Neonicotinoids are synthetic, nicotine-derived insecticides used for agricultural and household pest control. While highly effective at activating insect nicotinic receptors, many neonicotinoids are also capable of directly activating and/or modulating the activation of vertebrate nicotinic receptors. In this study, we have investigated the actions of the neonicotinoids clothianidin (CTD) and imidacloprid (IMI) on human neuronal α4β2 nicotinic acetylcholine receptors. The data demonstrate that the compounds are weak agonists of the human receptors with relative peak currents of 1–4 % of the response to 1 mM acetylcholine (ACh). Coapplication of IMI strongly inhibited currents elicited by ACh. From Schild plot analysis, we estimate that the affinity of IMI to the human α4β2 receptor is 18 µM. The application of low concentrations of CTD potentiated responses to low concentrations of ACh, suggesting that receptors occupied by one ACh and one CTD molecule have a higher gating efficacy than receptors with one ACh bound. Interestingly, subunit stoichiometry affected inhibition by CTD, with (α4)2(β2)3 receptors significantly more strongly inhibited than the (α4)3(β2)2 receptors.