Background: Visual cortex neurons often respond to stimuli very differently on repeated trials. This trial-by-trial variability is known to be correlated among nearby neurons. Our long-term goal is to quantitatively estimate neuronal response variability, using multi-channel local field potential (LFP) data from single trials.
Methods: Acute experiments were performed with anesthetized (Remifentanil, Propofol, nitrous oxide) and paralyzed (Gallamine Triethiodide) cats. Computer-controlled visual stimuli were displayed on a gamma-corrected CRT monitor. For the principal experiment, two kinds of visual stimuli were used: drifting sine-wave gratings, and a uniform mean-luminance gray screen. These two stimuli were each delivered monocularly for 100 sec in a random order, for 10 trials. Multi-unit activity (MUA) and LFP signals were extracted from broadband raw data acquired from Area 17 and 18 using A1X32 linear arrays (NeuroNexus) and the OpenEphys recording system. LFP signal processing was performed using Chronux, an open-source MATLAB toolbox. Current source density (CSD) analysis was performed on responses to briefly flashed full-field stimuli using the MATLAB toolbox, CSDplotter. The common response variability (global noise) of MUA was estimated using the model proposed by Scholvinck et al. [2015].
Results: On different trials, a given neuron responded with different firing to the same visual stimuli. Within one trial, a neuron’s firing rate also fluctuated across successive cycles of a drifting grating. When the animal was given extra anesthesia, neurons fired in a desynchronized pattern; with lighter levels of anesthesia, neuronal firing because more synchronized. By examining the cross-correlations of LFP signals recorded from different cortical layers, we found LFP signals could be divided to two groups: those recorded in layer IV and above, and those from layers V and VI. Within each group, LFP signals recorded by different channels are highly correlated. These two groups were observed in lighter and deeper anesthetized animals, also in sine-wave and uniform gray stimulus conditions. We also investigated correlations between LFP signals and global noise. Power in the LFP beta band was highly correlated with global noise, when animals were in deeper anesthesia.
Conclusions: Brain states contribute to variations in neuronal responses. Raw LFP correlation results suggest that we should analyze LFP data according to their laminar organization. Correlation of low-frequency LFP under deeper anesthesia with global noise gives us some insight to predict noise from single-trial data, and we hope to extend this analysis to lighter anesthesia in the future.
Background: The concept of stochastic facilitation suggests that the addition of precise amounts of white noise can improve the perceptibility of a stimulus of weak amplitude. We know from previous research that tactile and auditory noise can facilitate visual perception, respectively. Here we wanted to see if the effects of stochastic facilitation generalise to a reaction time paradigm, and if reaction times are correlated with tactile thresholds. We know that when multiple sensory systems are stimulated simultaneously, reaction times are faster than either stimulus alone, and also faster than the sum of reaction times (known as the race model).
Methods: Five participants were re-tested in five blocks each of which contained a different background noise levels, randomly ordered across sessions. At each noise level, they performed a tactile threshold detection task and a tactile reaction time task.
Results: Both tactile threshold and tactile reaction times were significantly affected by the background white noise. While the preferred amplitude for the white noise was different for every participant, the average lowest threshold was obtained with white noise presented binaurally at 70 db. The reaction times were analysed by fitting an ex-Gaussian, the sum of a Gaussian function and an exponential decay function. The white noise significantly affected the exponential parameter (tau) in a way that is compatible with the facilitation of thresholds.
Conclusions: We therefore conclude that multisensory reaction time facilitation can, at least in part, be explained by stochastic facilitation of the neural signals.
Background: Research suggests that the analysis of facial expressions by a healthy brain would take place approximately 170 ms after the presentation of a facial expression in the superior temporal sulcus and the fusiform gyrus, mostly in the right hemisphere. Some researchers argue that a fast pathway through the amygdala would allow automatic and early emotional treatment around 90 ms after stimulation. This treatment would be done subconsciously, even before this stimulus is perceived and could be approximated by presenting the stimuli quickly on the periphery of the fovea. The present study aimed to identify the neural correlates of a peripheral and simultaneous presentation of emotional expressions through a frequency tagging paradigm.
Methods: The presentation of emotional facial expressions at a specific frequency induces in the visual cortex a stable and precise response to the presentation frequency [i.e., a steady-state visual evoked potential (ssVEP)] that can be used as a frequency tag (i.e., a frequency-tag to follow the cortical treatment of this stimulus. Here, the use of different specific stimulation frequencies allowed us to label the different facial expressions presented simultaneously and to obtain a reliable cortical response being associated with (I) each of the emotions and (II) the different times of presentations repeated (1/0.170 ms =~5.8 Hz, 1/0.090 ms =~10.8 Hz). To identify the regions involved in emotional discrimination, we subtracted the brain activity induced by the rapid presentation of six emotional expressions of the activity induced by the presentation of the same emotion (reduced by neural adaptation). The results were compared to the hemisphere in which attention was sought, emotion and frequency of stimulation.
Results: The signal-to-noise ratio of the cerebral oscillations referring to the treatment of the expression of fear was stronger in the regions specific to the emotional treatment when they were presented in the subjects peripheral vision, unbeknownst to them. In addition, the peripheral emotional treatment of fear at 10.8 Hz was associated with greater activation within the Gamma 1 and 2 frequency bands in the expected regions (frontotemporal and T6), as well as desynchronization in the Alpha frequency bands for the temporal regions. This modulation of the spectral power is independent of the attentional request.
Conclusions: These results suggest that the emotional stimulation of fear presented in the peripheral vision and outside the attentional framework elicit an increase in brain activity, especially in the temporal lobe. The localization of this activity as well as the optimal stimulation frequency found for this facial expression suggests that it is treated by the fast pathway of the magnocellular layers.
Background: Our national collaborative research initiative is proposing to develop a common infrastructure for Rb research. We are proposing a novel in vivo Rb model using human Rb cells line.
Methods: The rabbit model has advantages over the mouse models: (I) the larger eye size of rabbits, similar to the human infant eye, permits a more accurate injection of the drugs and evaluation of methods of targeted intraocular drug delivery; (II) the rabbit model demonstrated similar fundus appearance and pathologic features to human Rb, including vitreous seeds of viable tumor when the retinal tumor is mid-sized, which are usually found in the late stage in mouse models. The lack of ability to eliminate vitreous seeds is a major reason of current treatment failures in Group C and D tumors; therefore, the rabbit model of Rb may be used as a model to evaluate the effectiveness and various routes of drug delivery.
Results: This is an implementation of an infrastructure for evaluating therapeutic targets. In addition, this finding enables a variety of pharmacokinetic studies, pharmacodynamic and toxicology studies for new therapeutic agents.
Conclusions: This infrastructure meets the growing concern of practitioners and researchers in the field. The common facility is easily accessible to all VHRN members on request, including requests from other sectors.
Background: Pericytes are contractile cells that wrap along the walls of capillaries. In the brain, pericytes play a crucial role in the regulation of capillary diameter and vascular blood flow in response to metabolic demand. During ischemia, it has been suggested that pericytes may constrict capillaries, and that pericytes remain constricted after reperfusion thus resulting in impaired blood flow.
Methods: Here, we used a mouse model of retinal ischemia based on ligation of the central retinal artery to characterize the role of pericytes on capillary constriction. Ischemia was induced in transgenic mice carrying the NG2 promoter driving red fluorescent protein expression to selectively visualize pericytes (line NG2:DsRed).Changes in retinal capillary diameter at 1 hr after ischemia were measured ex vivo in whole-mounted retinas from ischemic and control eyes (n=4–6/group) using a stereological approach. Vessels and pericytes were three-dimensionally reconstructed using IMARIS (Bitplane). Furthermore, we used a novel and minimally invasive two-photon microscopy approach that allowed live imaging of microvasculature changes in the retina.
Results: Our data show a generalized reduction in capillary diameter in ischemic retinas relative to sham-operated controls in all vascular plexus (ischemia: 4.7±0.2 μm, control: 5.2±0.2 μm, student’s t-test, P<0.001). Analysis of the number of capillary constrictions at pericyte locations, visualized in NG2:DsRed mice, demonstrated a substantial increase in ischemic retinas relative to the physiological capillary diameter reductions observed in controls (ischemia: 1,038±277 constrictions at pericyte locations, control: 60±36 constrictions at pericyte locations, student’s t-test, P<0.01). Live imaging using two-photon microscopy confirmed robust capillary constriction at the level of pericytes on retinal capillaries during ischemia (n=6–8/group).
Conclusions: Collectively, our data demonstrate that ischemia promotes rapid pericyte constriction on retinal capillaries causing major microvascular dysfunction in this tissue. To identify the molecular mechanisms underlying the pathological response of pericytes during ischemia, we are currently carrying out experiments in mice and zebrafish to modulate signaling pathways involved in calcium dynamics leading to contractility in these cells.
Background: Decrease of ocular blood flow has been linked to the pathogenesis of ocular diseases such as glaucoma and age-related macular degeneration. Current methods that measure the pulsatile blood flow have major limitations, including the assumption that ocular rigidity is the same in all eyes. Our group has recently developed a new method to measure the pulsatile choroidal volume change by direct visualization of the choroid with OCT imaging and automated segmentation. Our goal in this study is to describe the distribution of PCBF in a healthy Caucasian population.
Methods: Fifty-one subjects were recruited from the Maisonneuve-Rosemont Hospital Ophthalmology Clinic and underwent PCBF measurement in one eye. The distribution of PCBF in healthy eyes was assessed.
Results: The distribution of PCBF among the healthy eyes was found to be 3.94±1.70 μL with this technique.
Conclusions: This study demonstrates the normal range of PCBF values obtained in a healthy Caucasian population. This technique could be used for further investigation of choroid pulsatility and to study glaucoma pathophysiology.
Background: Zellweger spectrum disorder (ZSD) is an autosomal recessive disease caused by mutations in any one of 13 PEX genes whose protein products are required for peroxisome assembly. Retinopathy leading to blindness is one of the major handicaps faced by affected individuals, but treatment for this is supportive only. To test whether we could improve visual function in ZSD, we performed a proof-of-concept trial for PEX1 gene augmentation therapy using the Pex1-G844D mouse model, which bears the equivalent to a common human mutation. This model exhibits a gradual decline in scotopic ffERG response, an always residual photopic ffERG response, diminished visual acuity, and cone and bipolar cell anomalies.
Methods: We administered subretinal injections of a PEX1-containing viral vector (AAV8.CMV.hPEX1.HA) to 2 mouse cohorts of 5 or 9 weeks of age. A GFP-containing vector was used as a control in the contralateral eye of each animal. Efficient expression of the virus was confirmed by retinal histology/immunohistochemistry, and its ability to recover peroxisome import was confirmed in vitro. Preliminary ffERG and optokinetic (OKN) analyses were performed on a subset of animals at 8, 16, and 20 weeks after gene delivery. Final ffERG and OKN measures were performed when each cohort reached 32 weeks of age (23 or 27 weeks post injection).
Results: Preliminary ffERG and OKN analyses at 8 weeks post injection showed mildly better retinal response and visual acuity, respectively, in the PEX1-injected eyes, as did ffERG analysis when each cohort reached 25 weeks of age (16 or 20 weeks after gene delivery). This effect was more pronounced in the cohort treated at 5 weeks of age, when ffERG response is highest in Pex1-G844D mice. At 32 weeks of age, the ffERG response in the PEX1-injected eyes was double that of GFP-injected eyes, on average, though there was no change in OKN. Furthermore, in PEX1-injected eyes the photopic ffERG response improved over time, and the decline in scotopic b-wave amplitude was ameliorated compared to un-injected eyes.
Conclusions: AAV8.CMV.hPEX1.HA was subretinally delivered into the left eye of 5- and 9-week-old Pex1-G844D retina. Successful expression of the protein with no gross histologic side effect was observed. Neither the injection, nor exposure to the AAV8 capsid or the transgenic protein negatively altered the ERG or OKN response. At 5–6 months after gene delivery, therapeutic vector-treated eyes showed improved ERG compared to control eyes, on average, in both the “prevention” and “recovery” cohorts. This implies clinical potential of gene delivery to improve vision in patients with ZSD. Retinal immunohistochemistry (to visualize retinal cell types) and biochemical analyses will be performed on treated and untreated retinas, and may inform the mechanism of ERG improvement.
