Spatio-temporal activation patterns of neuronal population evoked by optostimulation and the comparison to electrical microstimulation.

Optostimulation and electrical microstimulation are well-established techniques that enable to artificially stimulate the brain. While the activation patterns evoked by microstimulation in cortical network are well characterized, much less is known for optostimulation. Specifically, the activation maps of neuronal population at the membrane potential level and direct measurements of these maps were barely reported. In addition, only a few studies compared the activation patterns evoked by microstimulation and optostimulation. In this study we addressed these issues by applying optostimulation in the barrel cortex of anesthetized rats after a short (ShortExp) or a long (LongExp) opsin expression time and compared it to microstimulation. We measured the membrane potential of neuronal populations at high spatial (meso-scale) and temporal resolution using voltage-sensitive dye imaging. Longer optostimulation pulses evoked higher neural responses spreading over larger region relative to short pulses. Interestingly, similar optostimulation pulses evoked stronger and more prolonged population response in the LongExp vs. the ShortExp condition. Finally, the spatial activation patterns evoked in the LongExp condition showed an intermediate state, with higher resemblance to the microstimulation at the stimulation site. Therefore, short microstimulation and optostimulation can induce wide spread activation, however the effects of optostimulation depend on the opsin expression time.

3K3A-Activated Protein C Inhibits Choroidal Neovascularization Growth and Leakage and Reduces NLRP3 Inflammasome, IL-1ß, and Inflammatory Cell Accumulation in the Retina.

3K3A-Activated Protein C (APC) is a recombinant variant of the physiological anticoagulant APC with cytoprotective properties and reduced bleeding risks. We studied the potential use of 3K3A-APC as a multi-target therapeutic option for choroidal neovascularization (CNV), a common cause of vision loss in age-related macular degeneration. CNV was induced by laser photocoagulation in a murine model, and 3K3A-APC was intravitreally injected. The impact of 3K3A-APC treatment on myeloid and microglia cell activation and recruitment and on NLRP3 inflammasome, IL-1ß, and VEGF levels was assessed using cryosection, retinal flat-mount immunohistochemistry and vascular imaging. Additionally, we evaluated the use of fluorescein angiography as a surrogate marker for in vivo evaluation of the efficacy of 3K3A-APC treatment against leaking CNV lesions. Our results demonstrated that 3K3A-APC treatment significantly reduced the accumulation and activation of myeloid cells and microglia in the CNV area and decreased the NLRP3 and IL-1ß levels at the CNV site and the surrounding retina. Furthermore, 3K3A-APC treatment resulted in leakage regression and CNV growth suppression. These findings indicate that the anti-inflammatory activities of 3K3A-APC contribute to CNV inhibition. Our study suggests the potential use of 3K3A-APC as a novel multi-target treatment for CNV.

Spatiotemporal patterns of population response in the visual cortex under isoflurane: from wakefulness to loss of consciousness.

Anesthetic drugs are widely used in medicine and research to mediate loss of consciousness (LOC). Isoflurane is a commonly used anesthetic drug; however, its effects on cortical sensory processing, in particular around LOC, are not well understood. Using voltage-sensitive dye imaging, we measured visually evoked neuronal population response from the visual cortex in awake and anesthetized mice at 3 increasing concentrations of isoflurane, thus controlling the level of anesthesia from wakefulness to deep anesthesia. At low concentration of isoflurane, the effects on neuronal measures were minor relative to the awake condition. These effects augmented with increasing isoflurane concentration, while around LOC point, they showed abrupt and nonlinear changes. At the network level, we found that isoflurane decreased the stimulus-evoked intra-areal spatial spread of local neural activation, previously reported to be mediated by horizontal connections, and also reduced intra-areal synchronization of neuronal population. The synchronization between different visual areas decreased with higher isoflurane levels. Isoflurane reduced the population response amplitude and prolonged their latencies while higher visual areas showed increased vulnerability to isoflurane concentration. Our results uncover the changes in neural activity and synchronization at isoflurane concentrations leading to LOC and suggest reverse hierarchical shutdown of cortical areas.

Spatio-temporal characteristics of population responses evoked by microstimulation in the barrel cortex.

Intra-cortical microstimulation (ICMS) is a widely used technique to artificially stimulate cortical tissue. This method revealed functional maps and provided causal links between neuronal activity and cognitive, sensory or motor functions. The effects of ICMS on neural activity depend on stimulation parameters. Past studies investigated the effects of stimulation frequency mainly at the behavioral or motor level. Therefore the direct effect of frequency stimulation on the evoked spatio-temporal patterns of cortical activity is largely unknown. To study this question we used voltage-sensitive dye imaging to measure the population response in the barrel cortex of anesthetized rats evoked by high frequency stimulation (HFS), a lower frequency stimulation (LFS) of the same duration or a single pulse stimulation. We found that single pulse and short trains of ICMS induced cortical activity extending over few mm. HFS evoked a lower population response during the sustained response and showed a smaller activation across time and space compared with LFS. Finally the evoked population response started near the electrode site and spread horizontally at a propagation velocity in accordance with horizontal connections. In summary, HFS was less effective in cortical activation compared to LFS although HFS had 5 fold more energy than LFS.