Key relationships between non-invasive functional neuroimaging and the underlying neuronal activity.

Functional neuroimaging using MRI relies on measurements of blood oxygen level-dependent (BOLD) signals from which inferences are made about the underlying neuronal activity. This is possible because neuronal activity elicits increases in blood flow via neurovascular coupling, which gives rise to the BOLD signal. Hence, an accurate interpretation of what BOLD signals mean in terms of neural activity depends on a full understanding of the mechanisms that underlie the measured signal, including neurovascular and neurometabolic coupling, the contribution of different cell types to local signalling, and regional differences in these mechanisms. Furthermore, the contributions of systemic functions to cerebral blood flow may vary with ageing, disease and arousal states, with regard to both neuronal and vascular function. In addition, recent developments in non-invasive imaging technology, such as high-field fMRI, and comparative inter-species analysis, allow connections between non-invasive data and mechanistic knowledge gained from invasive cellular-level studies. Considered together, these factors have immense potential to improve BOLD signal interpretation and bring us closer to the ultimate purpose of decoding the mechanisms of human cognition. This theme issue covers a range of recent advances in these topics, providing a multidisciplinary scientific and technical framework for future work in the neurovascular and cognitive sciences. This article is part of the theme issue 'Key relationships between non-invasive functional neuroimaging and the underlying neuronal activity'.

2015 Charles F. Prentice Medal Award Lecture: Neural Organization of Binocular Vision.

During a Research Career Development Award from the National Eye Institute, I spent a year at the University of Cambridge doing research with John Robson. The goal was to use a visual stimulation approach that had not been previously attempted, with the intention of exploring fundamental organization principles of the neural basis of binocular vision. The idea was to use sinusoidal gratings that drifted before both eyes such that the relative phase for one eye was fixed while that of the other was varied. This provided binocular stimuli of variable relative phase, i.e. retinal disparity, to enable testing of binocular response characteristics. We were able to obtain different types of disparity tuning functions for neurons in the primary visual cortex. This work, followed by extended investigations in Berkeley, provided basic information regarding response characteristics of simple and complex cells. We have also shown for monocular deprivation, an approximate model for human amblyopia, that many neurons remain connected to the deprived eye, as demonstrated with dichoptic activation. A selected portion of this work is described here.

Binocular function during unequal monocular input.

The fine task of stereoscopic depth discrimination in human subjects requires a functional binocular system. Behavioral investigations show that relatively small binocular abnormalities can diminish stereoscopic acuity. Clinical evaluations are consistent with this observation. Neurons in visual cortex represent the first stage of processing of the binocular system. Cells at this level are generally acutely sensitive to differences in relative depth. However, an apparent paradox in previous work demonstrates that tuning for binocular disparities remains relatively constant even when large contrast differences are imposed between left and right eye stimuli. This implies a range of neural binocular function that is at odds with behavioral findings. To explore this inconsistency, we have conducted psychophysical tests by which human subjects view vertical sinusoidal gratings drifting in opposite directions to left and right eyes. If the opposite drifting gratings are integrated in visual cortex, as wave theory and neurophysiological data predict, the subjects should perceive a fused stationary grating that is counter-phasing in place. However, this behavioral combination may not occur if there are differences in contrast and therefore signal strength between left and right eye stimuli. As expected for the control condition, our results show fused counter-phase perception for equal inter-ocular grating contrasts. Our experimental tests show a striking retention of counter-phase perception even for relatively large differences in inter-ocular contrast. This finding demonstrates that binocular integration, although relatively coarse, can occur during substantial differences in left and right eye signal strength.

Neural-metabolic coupling in the central visual pathway.

Studies are described which are intended to improve our understanding of the primary measurements made in non-invasive neural imaging. The blood oxygenation level-dependent signal used in functional magnetic resonance imaging (fMRI) reflects changes in deoxygenated haemoglobin. Tissue oxygen concentration, along with blood flow, changes during neural activation. Therefore, measurements of tissue oxygen together with the use of a neural sensor can provide direct estimates of neural-metabolic interactions. We have used this relationship in a series of studies in which a neural microelectrode is combined with an oxygen micro-sensor to make simultaneous co-localized measurements in the central visual pathway. Oxygen responses are typically biphasic with small initial dips followed by large secondary peaks during neural activation. By the use of established visual response characteristics, we have determined that the oxygen initial dip provides a better estimate of local neural function than the positive peak. This contrasts sharply with fMRI for which the initial dip is unreliable. To extend these studies, we have examined the relationship between the primary metabolic agents, glucose and lactate, and associated neural activity. For this work, we also use a Doppler technique to measure cerebral blood flow (CBF) together with neural activity. Results show consistent synchronously timed changes such that increases in neural activity are accompanied by decreases in glucose and simultaneous increases in lactate. Measurements of CBF show clear delays with respect to neural response. This is consistent with a slight delay in blood flow with respect to oxygen delivery during neural activation.This article is part of the themed issue 'Interpreting BOLD: a dialogue between cognitive and cellular neuroscience'.

Direction selectivity of neurons in the visual cortex is non-linear and lamina-dependent.

Neurons in the visual cortex are generally selective to direction of movement of a stimulus. Although models of this direction selectivity (DS) assume linearity, experimental data show stronger degrees of DS than those predicted by linear models. Our current study was intended to determine the degree of non-linearity of the DS mechanism for cells within different laminae of the cat's primary visual cortex. To do this, we analysed cells in our database by using neurophysiological and histological approaches to quantify non-linear components of DS in four principal cortical laminae (layers 2/3, 4, 5, and 6). We used a DS index (DSI) to quantify degrees of DS in our sample. Our results showed laminar differences. In layer 4, the main thalamic input region, most neurons were of the simple type and showed high DSI values. For complex cells in layer 4, there was a broad distribution of DSI values. Similar features were observed in layer 2/3, but complex cells were dominant. In deeper layers (5 and 6), DSI value distributions were characterized by clear peaks at high values. Independently of specific lamina, high DSI values were accompanied by narrow orientation tuning widths. Differences in orientation tuning for non-preferred vs. preferred directions were smallest in layer 4 and largest in layer 6. These results are consistent with a non-linear process of intra-cortical inhibition that enhances DS by selective suppression of neuronal firing for non-preferred directions of stimulus motion in a lamina-dependent manner. Other potential mechanisms are also considered.

Neurometabolic coupling between neural activity, glucose, and lactate in activated visual cortex.

Neural activity is closely coupled with energy metabolism but details of the association remain to be identified. One basic area involves the relationships between neural activity and the main supportive substrates of glucose and lactate. This is of fundamental significance for the interpretation of non-invasive neural imaging. Here, we use microelectrodes with high spatial and temporal resolution to determine simultaneous co-localized changes in glucose, lactate, and neural activity during visual activation of the cerebral cortex in the cat. Tissue glucose and lactate concentration levels are measured with electrochemical microelectrodes while neural spiking activity and local field potentials are sampled by a microelectrode. These measurements are performed simultaneously while neurons are activated by visual stimuli of different contrast levels, orientations, and sizes. We find immediate decreases in tissue glucose concentration and simultaneous increases in lactate during neural activation. Both glucose and lactate signals return to their baseline levels instantly as neurons cease firing. No sustained changes or initial dips in glucose or lactate signals are elicited by visual stimulation. However, co-localized measurements of cerebral blood flow and neural activity demonstrate a clear delay in the cerebral blood flow signal such that it does not correlate temporally with the neural response. These results provide direct real-time evidence regarding the coupling between co-localized energy metabolism and neural activity during physiological stimulation. They are also relevant to a current question regarding the role of lactate in energy metabolism in the brain during neural activation. Dynamic changes in energy metabolites can be measured directly with high spatial and temporal resolution by use of enzyme-based microelectrodes. Here, to examine neuro-metabolic coupling during brain activation, we use combined microelectrodes to simultaneously measure extracellular glucose, lactate, and neural responses in the primary visual cortex to visual stimulation. We demonstrate rapid decreases in glucose and increases in lactate during neural activation. Changes in glucose and lactate signals are transient and closely coupled with neuronal firing.

Transcranial Magnetic Stimulation Changes Response Selectivity of Neurons in the Visual Cortex.

BACKGROUND: Transcranial magnetic stimulation (TMS) is used to selectively alter neuronal activity of specific regions in the cerebral cortex. TMS is reported to induce either transient disruption or enhancement of different neural functions. However, its effects on tuning properties of sensory neurons have not been studied quantitatively. OBJECTIVE/HYPOTHESIS: Here, we use specific TMS application parameters to determine how they may alter tuning characteristics (orientation, spatial frequency, and contrast sensitivity) of single neurons in the cat's visual cortex. METHODS: Single unit spikes were recorded with tungsten microelectrodes from the visual cortex of anesthetized and paralyzed cats (12 males). Repetitive TMS (4 Hz, 4 s) was delivered with a 70 mm figure-8 coil. We quantified basic tuning parameters of individual neurons for each pre- and post-TMS condition. The statistical significance of changes for each tuning parameter between the two conditions was evaluated with a Wilcoxon signed-rank test. RESULTS: We generally find long-lasting suppression which persists well beyond the stimulation period. Pre- and post-TMS orientation tuning curves show constant peak values. However, strong suppression at non-preferred orientations tends to narrow the widths of tuning curves. Spatial frequency tuning exhibits an asymmetric change in overall shape, which results in an emphasis on higher frequencies. Contrast tuning curves show nonlinear changes consistent with a gain control mechanism. CONCLUSIONS: These findings suggest that TMS causes extended interruption of the balance between sub-cortical and intra-cortical inputs.

Spatiotemporal flow of information in the early visual pathway.

The spatial components of a visual scene are processed neurally in a sequence of coarse features followed by fine features. This coarse-to-fine temporal stream was initially considered to be a cortical function, but has recently been demonstrated in the dorsal lateral geniculate nucleus. The goal of this study was to test the hypothesis that coarse-to-fine processing is present at earlier stages of visual processing in the retinal ganglion cells that supply lateral geniculate nucleus (LGN) neurons. To compare coarse-to-fine processing in the cat's visual system, we measured the visual responses of connected neuronal pairs from the retina and LGN, and separate populations of cells from each region. We found that coarse-to-fine processing was clearly present at the ganglion cell layer of the retina. Interestingly, peak and high-spatial-frequency cutoff responses were higher in the LGN than in the retina, indicating that there was a progressive cascade of coarse-to-fine information from the retina to the LGN to the visual cortex. The analysis of early visual pathway receptive field characteristics showed that the physiological response interplay between the center and surround regions was consistent with coarse-to-fine features and may provide a primary role in the underlying mechanism. Taken together, the results from this study provided a framework for understanding the emergence and refinement of coarse-to-fine processing in the visual system.

Development of orientation tuning in simple cells of primary visual cortex.

Orientation selectivity and its development are basic features of visual cortex. The original model of orientation selectivity proposes that elongated simple cell receptive fields are constructed from convergent input of an array of lateral geniculate nucleus neurons. However, orientation selectivity of simple cells in the visual cortex is generally greater than the linear contributions based on projections from spatial receptive field profiles. This implies that additional selectivity may arise from intracortical mechanisms. The hierarchical processing idea implies mainly linear connections, whereas cortical contributions are generally considered to be nonlinear. We have explored development of orientation selectivity in visual cortex with a focus on linear and nonlinear factors in a population of anesthetized 4-wk postnatal kittens and adult cats. Linear contributions are estimated from receptive field maps by which orientation tuning curves are generated and bandwidth is quantified. Nonlinear components are estimated as the magnitude of the power function relationship between responses measured from drifting sinusoidal gratings and those predicted from the spatial receptive field. Measured bandwidths for kittens are slightly larger than those in adults, whereas predicted bandwidths are substantially broader. These results suggest that relatively strong nonlinearities in early postnatal stages are substantially involved in the development of orientation tuning in visual cortex.

Adaptation changes stereoscopic depth selectivity in visual cortex.

Exposure to specific visual stimuli causes a reduction in sensitivity to similar subsequent stimulation. This adaptation effect is observed behaviorally and for neurons in the primary visual cortex. Here, we explore the effects of adaptation on neurons that encode binocular depth discrimination in the cat's primary visual cortex. Our results show that neuronal preference for binocular depth is altered selectively with appropriate adaptation. At the preferred depth, adaptation causes substantial suppression of subsequent responses. Near the preferred depth, the same procedure causes a shift in depth preference. At the null depth, adaptation has little effect on binocular depth coding. These results demonstrate that prior exposure can change the depth selectivity of binocular neurons. The findings are relevant to the theoretical treatment of binocular depth processing. Specifically, the prevailing notion of binocular depth encoding based on the energy model requires modification.

Neurometabolic coupling differs for suppression within and beyond the classical receptive field in visual cortex.

Neurons in visual cortex exhibit two major types of stimulus elicited suppression. One, cross-orientation suppression, occurs within the classical receptive field (CRF) when an orthogonal grating is superposed on one at optimal orientation. The second, surround suppression, occurs when the size of an optimally oriented grating extends beyond the CRF. Previous proposals suggest that intracortical inhibition is responsible for surround suppression whereas feedforward processes may underlie cross-orientation suppression. To gain more insight concerning these types of suppression, we have included measurements of metabolic function in addition to neural responses. We made co-localized measurements of multiple unit neural activity and tissue oxygen concentrations in the striate cortex of anaesthetized cats while using visual stimuli to activate the two kinds of suppression. Results show that the amplitude of the initial negative oxygen response increases with stimulus size but neural responses decrease as size extends beyond the CRF. This shows that oxygen consumption increases with stimulus size regardless of reduced neural response. On the other hand, amplitudes of both the initial negative oxygen component and the neural responses are simultaneously attenuated by the orthogonal mask in cross-orientation suppression. These different neurometabolic response patterns are consistent with suggestions that the two types of suppressive processes arise from different neural mechanisms.

Neurometabolic coupling in the lateral geniculate nucleus changes with extended age.

Attempts have been made in various studies to identify and trace changes in function in the aging visual system. Some results are conflicting and we report here a unique approach in an attempt to resolve selected issues. We have estimated neurometabolic coupling in the central visual pathway in young and old cats. Our technique provides high resolution simultaneous measurements of neuronal activity and changes in concentration of tissue oxygen in the thalamus of young and old cats. Following visual stimulation, we find shorter latency and time to peak in tissue oxygen responses in old compared with young animals. Estimates of local activity induced initial negative oxygen response show substantial reductions in older animals. Measurements of neural activity in the form of multiple unit activity are similar in the two age groups. To investigate the mechanisms underlying the changes in tissue oxygen response in older animals, we measured vascular capillary density and found it to be substantially lower in old than that in young animals. Together, these findings suggest that the changes in metabolic responses with age may be largely accounted for by alterations in the cerebral microvasculature rather than by changes in neural activity.

Pulfrich phenomena are coded effectively by a joint motion-disparity process.

Pulfrich phenomena are a class of depth illusions generated by an interocular time delay. This may be demonstrated with continuously moving stimuli, stroboscopic displays undergoing apparent motion, or dynamic noise patterns. Previous studies suggest that neurons jointly tuned to motion and disparity may be responsible for the phenomena. Model cells with such joint coding can explain all Pulfrich phenomena in a unified way (N. Qian & R. A. Andersen, 1997). However, the joint-coding idea has been challenged by recent models (J. C. Read & B. G. Cumming, 2005a, 2005c) that focus on the S shaped functions of perceived disparity in stroboscopic Pulfrich effect (M. J. Morgan, 1979). Here we demonstrate fundamental problems with the recent models in terms of causality, physiological plausibility, and definitions for joint and separate coding, and we compare the two coding schemes under physiologically plausible assumptions. We show that joint coding of disparity and either unidirectional or bidirectional motion selectivity can account for the S curves, but unidirectional selectivity is required to explain direction-depth contingency in Pulfrich effects. In contrast, separate coding can explain neither the S curves nor the direction-depth contingency. We conclude that Pulfrich phenomena are logically accounted for by joint encoding of unidirectional-motion and disparity.

State-dependent variability of neuronal responses to transcranial magnetic stimulation of the visual cortex.

Electrical brain stimulation is a promising tool for both experimental and clinical applications. However, the effects of stimulation on neuronal activity are highly variable and poorly understood. To investigate the basis of this variability, we performed extracellular recordings in the visual cortex following application of transcranial magnetic stimulation (TMS). Our measurements of spiking and local field potential activity exhibit two types of response patterns which are characterized by the presence or absence of spontaneous discharge following stimulation. This variability can be partially explained by state-dependent effects, in which higher pre-TMS activity predicts larger post-TMS responses. These results reveal the possibility that variability in the neural response to TMS can be exploited to optimize the effects of stimulation. It is conceivable that this feature could be utilized in real time during the treatment of clinical disorders.

Contrast sensitivity is enhanced by expansive nonlinear processing in the lateral geniculate nucleus.

The firing rates of neurons in the central visual pathway vary with stimulus strength, but not necessarily in a linear manner. In the contrast domain, the neural response function for cells in the primary visual cortex is characterized by expansive and compressive nonlinearities at low and high contrasts, respectively. A compressive nonlinearity at high contrast is also found for early visual pathway neurons in the lateral geniculate nucleus (LGN). This mechanism affects processing in the visual cortex. A fundamentally related issue is the possibility of an expansive nonlinearity at low contrast in LGN. To examine this possibility, we have obtained contrast-response data for a population of LGN neurons. We find for most cells that the best-fit function requires an expansive component. Additionally, we have measured the responses of LGN neurons to m-sequence white noise and examined the static relationship between a linear prediction and actual spike rate. We find that this static relationship is well fit by an expansive nonlinear power law with average exponent of 1.58. These results demonstrate that neurons in early visual pathways exhibit expansive nonlinear responses at low contrasts. Although this thalamic expansive nonlinearity has been largely ignored in models of early visual processing, it may have important consequences because it potentially affects the interpretation of a variety of visual functions.

High-resolution neurometabolic coupling in the lateral geniculate nucleus.

The relationships between neural and metabolic processes in activated brain regions are central to the interpretation of noninvasive imaging. To examine this relationship, we have used a specialized sensor to measure simultaneously tissue oxygen changes and neural activity in colocalized regions of the cat's lateral geniculate nucleus (LGN). Previous work with this sensor has shown that a decrease or increase in tissue oxygen can be elicited by selective control of the location and extent of neural activation in the LGN. In the current study, to evaluate the temporal integration and homogeneity of neurometabolic coupling, we have determined the relationship between multiunit extracellular neural activity and tissue oxygen responses to visual stimuli of various durations and contrasts. Our results show that the negative but not the positive oxygen response changes in an approximately linear manner with stimulus duration. The relationship between the negative oxygen response and neural activity is relatively constant with stimulus duration. Moreover, both negative and positive oxygen responses saturate at high stimulus contrast levels. Coupling between neural activity and negative oxygen responses is well described by a power law function. These results help elucidate differences between the initial negative and subsequent positive metabolic responses and may be directly relevant to questions concerning brain mapping with functional magnetic resonance imaging.

Neurometabolic coupling in cerebral cortex reflects synaptic more than spiking activity.

In noninvasive neuroimaging, neural activity is inferred from local fluctuations in deoxyhemoglobin. A fundamental question of functional magnetic resonance imaging (fMRI) is whether the inferred neural activity is driven primarily by synaptic or spiking activity. The answer is critical for the interpretation of the blood oxygen level-dependent (BOLD) signal in fMRI. Here, we have used well-established visual-system circuitry to create a stimulus that elicits synaptic activity without associated spike discharge. In colocalized recordings of neural and metabolic activity in cat primary visual cortex, we observed strong coupling between local field potentials (LFPs) and changes in tissue oxygen concentration in the absence of spikes. These results imply that the BOLD signal is more closely coupled to synaptic activity.

Transcranial magnetic stimulation elicits coupled neural and hemodynamic consequences.

Transcranial magnetic stimulation (TMS) is an increasingly common technique used to selectively modify neural processing. However, application of TMS is limited by uncertainty concerning its physiological effects. We applied TMS to the cat visual cortex and evaluated the neural and hemodynamic consequences. Short TMS pulse trains elicited initial activation (approximately 1 minute) and prolonged suppression (5 to 10 minutes) of neural responses. Furthermore, TMS disrupted the temporal structure of activity by altering phase relationships between neural signals. Despite the complexity of this response, neural changes were faithfully reflected in hemodynamic signals; quantitative coupling was present over a range of stimulation parameters. These results demonstrate long-lasting neural responses to TMS and support the use of hemodynamic-based neuroimaging to effectively monitor these changes over time.

Spatial frequency-specific contrast adaptation originates in the primary visual cortex.

Adaptation to a high-contrast grating stimulus causes reduced sensitivity to subsequent presentation of a visual stimulus with similar spatial characteristics. This behavioral finding has been attributed by neurophysiological studies to processes within the visual cortex. However, some evidence indicates that contrast adaptation phenomena are also found in early visual pathways. Adaptation effects have been reported in retina and lateral geniculation nucleus (LGN). It is possible that these early pathways could be the physiological origin of the cortical adaptation effect. To study this, we recorded from single neurons in the cat's LGN. We find that contrast adaptation in the LGN, unlike that in the visual cortex, is not spatial frequency specific, i.e., adaptation effects apply to a broad range of spatial frequencies. In addition, aside from the amplitude attenuation, the shape of spatial frequency tuning curves of LGN cells is not affected by contrast adaptation. Again, these findings are unlike those found for cells in the visual cortex. Together, these results demonstrate that pattern specific contrast adaptation is a cortical process.

Analysis of oxygen metabolism implies a neural origin for the negative BOLD response in human visual cortex.

The sustained negative blood oxygenation level-dependent (BOLD) response in functional MRI is observed universally, but its interpretation is controversial. The origin of the negative response is of fundamental importance because it could provide a measurement of neural deactivation. However, a substantial component of the negative response may be due to a non-neural hemodynamic artifact. To distinguish these possibilities, we have measured evoked BOLD, cerebral blood flow (CBF), and oxygen metabolism responses to a fixed visual stimulus from two different baseline conditions. One is a normal resting baseline, and the other is a lower baseline induced by a sustained negative response. For both baseline conditions, CBF and oxygen metabolism responses reach the same peak amplitude. Consequently, evoked responses from the negative baseline are larger than those from the resting baseline. The larger metabolic response from negative baseline presumably reflects a greater neural response that is required to reach the same peak amplitude as that from resting baseline. Furthermore, the ratio of CBF to oxygen metabolism remains approximately the same from both baseline states (approximately 2:1). This tight coupling between hemodynamic and metabolic components implies that the magnitude of any hemodynamic artifact is inconsequential. We conclude that the negative response is a functionally significant index of neural deactivation in early visual cortex.