Of Intrinsic Signals Research Articles

Mapping mesoscale cortical connectivity in monkey sensorimotor cortex with optical imaging and microstimulation.

To map in vivo cortical circuitry at the mesoscale, we applied a novel approach to map interareal functional connectivity. Electrical intracortical microstimulation (ICMS) in conjunction with optical imaging of intrinsic signals (OIS) was used map functional connections in somatosensory cortical areas in anesthetized squirrel monkeys. ICMS produced activations that were focal and that displayed responses which were stimulation intensity dependent. ICMS in supragranular layers of Brodmann Areas 3b, 1, 2, 3a, and M1 evoked interareal activation patterns that were topographically appropriate and appeared consistent with known anatomical connectivity. Specifically, ICMS revealed Area 3b connections with Area 1; Area 1 connections with Areas 2 and 3a; Area 2 connections with Areas 1, 3a, and M1; Area 3a connections with Areas M1, 1, and 2; and M1 connections with Areas 3a, 1, and 2. These somatosensory connectivity patterns were reminiscent of feedforward patterns observed anatomically, although feedback contributions are also likely present. Further consistent with anatomical connectivity, intra-areal and intra-areal patterns of activation were patchy with patch sizes of 200-300 μm. In summary, ICMS with OIS is a novel approach for mapping interareal and intra-areal connections in vivo. Comparisons with feedforward and feedback anatomical connectivity are discussed.

Comparison of cortical responses to the activation of retina by visual stimulation and transcorneal electrical stimulation

BackgroundElectrical stimulation has been widely used in many ophthalmic diseases to modulate neuronal activities or restore partial visual function. Due to the different processing pathways and mechanisms, responses to visual and electrical stimulation in the primary visual cortex and higher visual areas might be different. This differences would shed some light on the properties of cortical responses evoked by electrical stimulation. ObjectiveThis study's goal was to directly compare the cortical responses evoked by visual and electrical stimulation and investigate the cortical processing of visual information and extrinsic electrical signal. MethodsOptical imaging of intrinsic signals (OIS) was used to probe the cortical hemodynamic responses in 11 cats. Transcorneal electrical stimulation (TES) through an ERG-jet contact lens electrode was used to activate visual cortices. Full-field and peripheral drifting gratings were used as the visual stimuli. ResultsThe response latency evoked by TES was shorter than that responding to visual stimulation (VS). Cortical responses evoked by VS were retinotopically organized, which was consistent with previous studies. On the other hand, the cortical region activated by TES was preferentially located in the secondary visual cortex (Area 18), while the primary visual cortex (Area 17) was activated by a higher current intensity. Compared with the full-field VS, the cortical response in Area 18 to TES with a current intensity above 1.2 mA was significantly stronger. ConclusionAccording to our results, we provided some evidence that the cortical processing of TES was influenced by the distribution of the electrical field in the retina and the activating threshold of different retinal ganglion cells.

Improved Recordings of the Optical Intrinsic Signals in the Neonatal Rat Barrel Cortex

Optical imaging of intrinsic signals (OIS) is widely used for the functional cortical mapping in vivo. Recently, OIS is also implemented for the functional mapping in the neonatal rat barrel cortex. However, the OIS is characterized by relatively low signal to noise ratio (SNR). Here, we determined parameters for post hoc data analysis that allowed improving OIS mapping and analysis in the developing rat barrel cortex in vivo. We found that application of spatial Gaussian filtering with sigma of 1 px increases the OIS SNR almost twofold. Additional light correction and low-pass temporal filtering with 1 s window size resulted in further improvement of the OIS SNR. Thus, the proposed digital filtering can substantially improve quality of the OIS recordings in the developing somatosensory cortex.

Optical imaging of visual cortical responses evoked by transcorneal electrical stimulation with different parameters.

The use of phosphenes evoked by transcorneal electrical stimulation (TcES) has been proposed as a means of residual visual function evaluation and candidate selection before implantation of retinal prostheses. Compared to the subjective measures, measurement of neuronal activity in visual cortex can objectively and quantitatively explore their response properties to electrical stimulation. The purpose of this study was to investigate systematically the properties of cortical responses evoked by TcES. The visual cortical responses were recorded using a multiwavelength optical imaging of intrinsic signals (OIS) combining with electrophysiological recording by a multichannel electrode array. The effects of different parameters of TcES on cortical responses, including the changes of hemoglobin oxygenation and cerebral blood volume, were examined. We found consistent OIS activation regions in visual cortex after TcES, which also showed strong evoked field potentials according to electrophysiological results. The OIS response regions were located mainly in cortical areas representing peripheral visual field. The extent of activation areas and strength of intrinsic signals were increased with higher current intensities and longer pulse widths, and the largest responses were acquired in the frequency range 10 to 20 Hz. Use of TcES through the ERG-jet corneal electrode may preferentially activate peripheral retina. Revealing the hemodynamic changes in visual cortex occurred after electrical stimulation can contribute to comprehension of neurophysiological underpinnings underlying prosthetic vision. This study provided an objective foundation for optimizing parameters of TcES and would bring considerable benefits in the application of TcES for assessment and screening in patients.

Flavoprotein autofluorescence imaging of visual system activity in zebra finches and mice.

Large-scale brain activity patterns can be visualized by optical imaging of intrinsic signals (OIS) based on activity-dependent changes in the blood oxygenation level. Another method, flavoprotein autofluorescence imaging (AFI), exploits the mitochondrial flavoprotein autofluorescence, which is enhanced during neuronal activity. In birds, topographic mapping of visual space has been shown in the visual wulst, the avian homologue of the mammalian visual cortex by using OIS. We here applied the AFI method to visualize topographic maps in the visual wulst because with OIS, which depends on blood flow changes, blood vessel artifacts often obscure brain activity maps. We then compared both techniques quantitatively in zebra finches and in C57Bl/6J mice using the same setup and stimulation conditions. In addition to experiments with craniotomized animals, we also examined mice with intact skull (in zebra finches, intact skull imaging is not feasible probably due to the skull construction). In craniotomized animals, retinotopic maps were obtained by both methods in both species. Using AFI, artifacts caused by blood vessels were generally reduced, the magnitude of neuronal activity significantly higher and the retinotopic map quality better than that obtained by OIS in both zebra finches and mice. In contrast, our measurements in non-craniotomized mice did not reveal any quantitative differences between the two methods. Our results thus suggest that AFI is the method of choice for investigations of visual processing in zebra finches. In mice, however, if researchers decide to use the advantages of imaging through the intact skull, they will not be able to exploit the higher signals obtainable by the AFI-method.

Optical imaging of cortical networks via intracortical microstimulation

Understanding cortical organization is key to understanding brain function. Distinct neural networks underlie the functional organization of the cerebral cortex; however, little is known about how different nodes in the cortical network interact during perceptual processing and motor behavior. To study cortical network function we examined whether the optical imaging of intrinsic signals (OIS) reveals the functional patterns of activity evoked by electrical cortical microstimulation. We examined the effects of current amplitude, train duration, and depth of cortical stimulation on the hemodynamic response to electrical microstimulation (250-Hz train, 0.4-ms pulse duration) in anesthetized New World monkey somatosensory cortex. Electrical stimulation elicited a restricted cortical response that varied according to stimulation parameters and electrode depth. Higher currents of stimulation recruited more areas of cortex than smaller currents. The largest cortical responses were seen when stimulation was delivered around cortical layer 4. Distinct local patches of activation, highly suggestive of local projections, around the site of stimulation were observed at different depths of stimulation. Thus we find that specific electrical stimulation parameters can elicit activation of single cortical columns and their associated columnar networks, reminiscent of anatomically labeled networks. This novel functional tract tracing method will open new avenues for investigating relationships of local cortical organization.

Cerebral Artery–Vein Separation Using 0.1-Hz Oscillation in Dual-Wavelength Optical Imaging

We present a novel artery-vein separation method using 0.1-Hz oscillation at two wavelengths with optical imaging of intrinsic signals (OIS). The 0.1-Hz oscillation at a green light wavelength of 546 nm exhibits greater amplitude in arteries than in veins and is primarily caused by vasomotion, whereas the 0.1-Hz oscillation at a red light wavelength of 630 nm exhibits greater amplitude in veins than in arteries and is primarily caused by changes of deoxyhemoglobin concentration. This spectral feature enables cortical arteries and veins to be segmented independently. The arteries can be segmented on the 0.1-Hz amplitude image at 546 nm using matched filters of a modified dual Gaussian model combining with a single Gaussian model. The veins are a combination of vessels segmented on both amplitude images at the two wavelengths using multiscale matched filters of single Gaussian model. Our method can separate most of the thin arteries and veins from each other, especially the thin arteries with low contrast in raw gray images. In vivo OIS experiments demonstrate the separation ability of the 0.1-Hz based segmentation method in cerebral cortex of eight rats. Two validation studies were undertaken to evaluate the performance of the method by quantifying the arterial and venous length based on a reference standard. The results indicate that our 0.1-Hz method is very effective in separating both large and thin arteries and veins regardless of vessel crossover or overlapping to great extent in comparison with previous methods.

The biological effect of contralateral forepaw stimulation in rat focal cerebral ischemia: a multispectral optical imaging study

Our group has already published the possible neuroprotective effect of contralateral forepaw stimulation in temporary focal ischemia in a study. However, the background is still unclear. In the present study we investigated the possible mechanism by monitoring focal ischemia with multispectral [laser speckle, imaging of intrinsic signals (OIS)] imaging. Sprague–Dawley rats were prepared using 1.2% isoflurane anesthesia. The middle cerebral artery was occluded by photothrombosis (4 mW) and the common carotid artery was ligated permanently. Physiological variables were constantly monitored during the experiment. A 6 × 6 mm area centered 3 mm posterior and 4 mm lateral to Bregma was thinned for laser speckle and OIS imaging. Nine circular regions of interests (0.3 mm in diameter) were evenly spaced on the speckle contrast image for the analysis of peri-infarct flow transients, blood flow, and metabolic changes. Both the sham (n = 7) and forepaw-stimulated animals (n = 7) underwent neurological examinations 24 h after ischemia at which point all animals were sacrificed and the infarct size was determined by triphenyltetrazolium chloride. The physiological variables were in normal range and the experimental protocol did not cause significant differences between groups. Both the neurological scores (sham: 3.6 ± 1.7, stimulated: 4.3 ± 1.4) and the infarct volume (sham: 124 ± 39 mm3, stimulated: 147 ± 47 mm3) did not show significant differences between groups. The forepaw stimulation did not increase the intra-ischemic flow neither over the penumbral or the peri-ischemic area. However, the hemoglobin transients related metabolic load (CMRO2) was significantly lower (p < 0.001) while the averaged number of hyperemic flow transients were significantly (p = 0.013) higher in the forepaw (sham: 3.5 ± 2.2, stimulated: 7.0 ± 2.3) stimulated animals.

Neurovascular uncoupling: Much ado about nothing

The vast majority of human fMRI studies measure Blood-Oxygen-Level-Dependent (BOLD) contrast, which reflects regional changes in cerebral blood flow (CBF), cerebral blood volume (CBV) and blood oxygenation; all three vascular responses reflect local increases in neural activity (Logothetis and Wandell, 2004). Understanding the exact mechanism (often referred to as neurovascular coupling) by means of which changes in neural activity alter hemodynamics is obviously of paramount importance for the meaningful interpretation of fMRI results. Not surprisingly, over the last decade an increasing number of researchers investigated the neurovascular coupling by combining fMRI with electroencephalography (EEG) or magnetoencephalography (MEG) in humans, e.g. (Dale and Halgren, 2001) as well as with intracortical recordings in animals (Logothetis et al., 2001; Goense and Logothetis, 2008; Logothetis, 2008; Rauch et al., 2008). This neurovascular coupling can also be studied with the optical imaging of intrinsic signals (OIS) (Bonhoeffer and Grinvald, 1996), an excellent invasive method of high spatiotemporal resolution that can measure changes in oxygenation and/or blood volume.

Trial-by-trial relationship between neural activity, oxygen consumption, and blood flow responses

Trial-by-trial variability in local field potential (LFP), tissue partial pressure of oxygen (PO2), cerebral blood flow (CBF), and deoxyhemoglobin-weighted optical imaging of intrinsic signals (OIS) were tested in the rat somatosensory cortex while fixed electrical forepaw stimulation (1.0-ms pulses with amplitude of 1.2 mA at a frequency of 6 Hz) was repeatedly applied. The changes in the cerebral metabolic rate of oxygen (CMRO2) were also evaluated using a hypotension condition established by our group based on the administration of a vasodilator. Under normal conditions, CBF, PO2, and OIS showed positive signal changes (48%, 32%, and 0.42%, respectively) following stimulation. Over multiple trials, the CBF responses were well correlated with the integral of the LFP amplitudes (∑LFP) (Rmean=0.78), whereas a lower correlation was found between PO2 and ∑LFP (Rmean=0.60) and between OIS and ∑LFP (Rmean=0.54). Under the hypotension condition the LFP responses were preserved, but the CBF responses were suppressed and the PO2 and OIS changes were negative (−12% and −0.28%, respectively). In this condition, the trial-by-trial variations in PO2 and OIS were well correlated with the variability in ∑LFPs (Rmean=−0.77 and −0.76, respectively), indicating a single trial coupling between CMRO2 changes and ∑LFP. These findings show that CBF and CMRO2 signals are more directly correlated with neural activity compared to blood oxygen-sensitive methods such as OIS and BOLD fMRI.

Perfusion‐based fMRI: Insights from animal models

Modern functional neuroimaging techniques, including functional magnetic resonance imaging (fMRI), positron emission tomography (PET), and optical imaging of intrinsic signals (OIS), rely on a tight coupling between neural activity and cerebral blood flow (CBF) to visualize brain activity using CBF as a surrogate marker. Because CBF is a uniquely defined physiological parameter, fMRI techniques based on CBF contrast have the advantage of being specific to tissue signal change, and the potential to provide more direct and quantitative measures of brain activation than blood oxygenation level-dependent (BOLD)- or cerebral blood volume (CBV)-based techniques. The changes in CBF elicited by increased neural activity are an excellent index of the magnitude of electrical activity. Increases in CBF are more closely localized to the foci of increased electrical activity, and occur more promptly to the stimulus than BOLD- or CBV-based contrast. In addition, CBF-based fMRI is less affected by confounds from venous drainage common to BOLD. Animal studies of brain activation have yielded considerable insights into the advantages of CBF-based fMRI. Based on results provided by animal studies, CBF fMRI may offer a means of better assessing the magnitude, spatial extent, and temporal response of neural activity, and may be more specific to tissue state. These properties are expected to be particularly useful for longitudinal and quantitative fMRI studies.

Isofrequency Band-like Zones of Activation Revealed by Optical Imaging of Intrinsic Signals in the Cat Primary Auditory Cortex

Neurons of similar frequency preference are arranged in isofrequency bands (IFBs) across the primary auditory cortex (AI) of many mammals. Across the AI of the cat, one of the most frequently studied species for auditory anatomy and function, we demonstrate IFB-like responses using optical imaging of intrinsic signals (OIS). Optically defined activations were extensively elongated along the dorsoventral axis of AI (the ratio of the major and minor axes was approximately 2:1), and systematically shifted as a function of stimulus frequency. The elongation of this IFB-like zone was more conspicuous at higher frequencies. In the ventral sector of the imaged field, the IFB-like zones of activation evoked at different pure tone frequencies tended to overlap extensively. Electrophysiological recording from loci within the optically defined zones of activation revealed matched responses to the frequencies used for optical imaging at 65% of these loci. The dorsoventral orientation of these zones of activation was also closely matched with the orientation of tangentially spreading intrinsic axon terminals, as revealed anatomically. The visualization of IFB-like architecture and tonotopic organization by OIS provides a basic framework for investigating the relationships of different spectral channels and between multiple acoustic parameters at a neuronal population level.

Precise placement of multiple electrodes into functionally predefined cortical locations

Current research on topics such as effective connectivity, neuronal coding strategy or signal propagation in the central nervous system requires simultaneous recordings from multiple sites within functionally grouped but topologically distributed neuronal clusters. We have addressed this issue by characterization of the cortical functional architecture using optical imaging of intrinsic signals (OI) and subsequent placement of multiple, individually adjustable electrodes into pre-selected domains. In order to achieve maximum precision and flexibility for the positioning of electrodes, a plastic cylinder containing channels of an extremely high aspect ratio (density >20 channels/mm 2) was fixed above the cortex and individual channel positions were superimposed onto the functional maps of orientation columns obtained previously with OI. Subsequently, channels corresponding to the desired locations in the functional map were used as guide tubes for electrode insertion. The spatial precision of this approach was in the range of 100 μm and experiments in cat primary visual cortex revealed a close correlation between the desired and the actually recorded orientation preferences of the targeted columns. The method is applicable to all cortical areas in which OI is feasible and offers a high degree of flexibility with respect to the number and geometry of applicable probes. It is, thus, an excellent tool for studying distributed codes and interactions between multiple predefined recording sites.

Evaluation of coupling between optical intrinsic signals and neuronal activity in rat somatosensory cortex

We investigated the coupling between perfusion-related brain imaging signals and evoked neuronal activity using optical imaging of intrinsic signals (OIS) at 570 and 610 nm. OIS at 570 nm reflects changes in cerebral blood volume (CBV), and the 610 nm response is related to hemoglobin oxygenation changes. We assessed the degree to which these components of the hemodynamic response were coupled to neuronal activity in rat barrel, hindpaw, and forepaw somatosensory cortex by simultaneously recording extracellular evoked field potentials (EPs) and OIS while varying stimulation frequency. In all stimulation paradigms, 10 Hz stimulation evoked the largest optical and electrophysiological responses. Across all animals, the 610 late phase and 570 responses correlated linearly with ΣEP ( P < 0.05) during both whisker deflection and electrical hindpaw stimulation, but the 610 early phase did not (whisker P = 0.27, hindpaw P = 0.28). The signal-to-noise ratio (SNR) of the 610 early phase (whisker 3.1, hindpaw 5.3) was much less than that for the late phase (whisker 14, hindpaw 51) and 570 response (whisker 11, hindpaw 46). During forepaw stimulation, however, the 610 early phase had a SNR (17) higher than that during hindpaw stimulation and correlated well with neuronal activity ( P < 0.05). We conclude that the early deoxygenation change does not correlate consistently with neuronal activity, possibly because of its low SNR. The robust CBV-related response, however, has a high SNR and correlates well with evoked cortical activity.

Intraoperative optical intrinsic signal imaging: a clinical tool for functional brain mapping

Optical imaging of intrinsic signals (OIS) is a well-established neuroimaging modality by which functional cortical activity is mapped by detecting activity-related changes in cortical light reflectance. Light reflectance changes are detected by a charged-coupled device camera that captures images of the exposed cortex both at rest and during activity. Although to date OIS has only been used for research purposes, intraoperative OIS (iOIS) holds promise as a clinical mapping tool. In general, iOIS demonstrates good spatial correlation with electrocortical stimulation mapping (ECSM) and other electrophysiological modalities. Additionally, iOIS offers high spatial resolution (in microns), does not make contact with the surface of the brain, and introduces no potentially harmful compounds. Moreover, mapping is relatively rapid. The authors review the potential contribution of iOIS to the intraoperative environment. Specifically, they review iOIS methodology, discuss signal origin, compare OIS with other functional mapping modalities, and explain its potential benefits and limitations. They propose that iOIS may, in the future, be used in conjunction with ECSM to improve the resolution and accuracy of intraoperative mapping, decrease total time of intraoperative mapping, and possibly improve neurological outcomes. Additional studies will be required to quantify the sensitivity and specificity of optical maps relative to ECSM before it can be implemented clinically.

Brief visual stimulation allows mapping of ocular dominance in visual cortex using fMRI.

We have used high spatial resolution (0.55 mm x 0.55 mm) functional magnetic resonance imaging (fMRI) to show that when stimulus duration is brief (<6 sec), the hyperoxic hemodynamic response to neural activity can resolve the columnar architecture of ocular dominance within the primary visual cortex of humans. Our fMRI maps of ocular dominance columns are strikingly similar in appearance, size, and orientation to those reported in the literature using optical imaging of intrinsic signals (OIS) in animal cortex and histology of post-mortem human specimens. We also demonstrate that under brief visual stimulation conditions, our results are consistent over repeated experiments. This is not the case for long duration stimuli (> or = 10 sec). A simulated random data set exhibited the same response properties as maps obtained when using these prolonged visual stimuli. Our results suggest that brief visual stimulation is essential for fMRI to successfully resolve ocular dominance columns using the hyperoxic phase of the hemodynamic response to neural activity at our prescribed spatial resolution.