Neurophysiology of the BOLD fMRI signal in awake monkeys.

BACKGROUND: Simultaneous intracortical recordings of neural activity and blood-oxygen-level-dependent (BOLD) functional magnetic resonance imaging (fMRI) in primary visual cortex of anesthetized monkeys demonstrated varying degrees of correlation between fMRI signals and the different types of neural activity, such as local field potentials (LFPs), multiple-unit activity (MUA), and single-unit activity (SUA). One important question raised by the aforementioned investigation is whether the reported correlations also apply to alert subjects. RESULTS: Monkeys were trained to perform a fixation task while stimuli within the receptive field of each recording site were used to elicit neural responses followed by a BOLD response. We show -- also in alert behaving monkeys -- that although both LFP and MUA make significant contributions to the BOLD response, LFPs are better and more reliable predictors of the BOLD signal. Moreover, when MUA responses adapt but LFP remains unaffected, the BOLD signal remains unaltered. CONCLUSIONS: The persistent coupling of the BOLD signal to the field potential when LFP and MUA have different time evolutions suggests that BOLD is primarily determined by the local processing of inputs in a given cortical area. In the alert animal the largest portion of the BOLD signal's variance is explained by an LFP range (20-60 Hz) that is most likely related to neuromodulation. Finally, the similarity of the results in alert and anesthetized subjects indicates that at least in V1 anesthesia is not a confounding factor. This enables the comparison of human fMRI results with a plethora of electrophysiological results obtained in alert or anesthetized animals.

Functional magnetic resonance imaging of awake behaving macaques.

In recent years, more and more laboratories have developed functional Magnetic Resonance Imaging (fMRI) for awake non-human primates. This research is essential to provide a link between non-invasive hemodynamic signals recorded in the human brain and the vast body of knowledge gained from invasive electrophysiological studies in monkeys. Given that their brain structure is so closely related to that of humans and that monkeys can be trained to perform complicated behavioral tasks, results obtained with monkey fMRI and electrophysiology can be compared to fMRI results obtained in humans, and provide information crucial to a better understanding of the mechanisms by which different cortical areas perform their functions in the human brain. However, despite that the first publications on fMRI in awake behaving macaques appeared approximately 10 years ago (Logothetis et al. (1999) [1], Stefanacci et al. (1998) [2], Dubowitz et al. (1998) [3]), relatively few laboratories perform such experiments routinely, a sign of the significant technical difficulties that must be overcome. The higher spatial resolution required because of the animal's smaller brain results in poorer signal-to-noise ratios than in human fMRI, which is further compounded by problems due to animal motion. Here, we discuss the specific challenges and benefits of fMRI in the awake monkey and review the methodologies and strategies for scanning behaving macaques.

Fine-scale spatial organization of face and object selectivity in the temporal lobe: do functional magnetic resonance imaging, optical imaging, and electrophysiology agree?

The spatial organization of the brain's object and face representations in the temporal lobe is critical for understanding high-level vision and cognition but is poorly understood. Recently, exciting progress has been made using advanced imaging and physiology methods in humans and nonhuman primates, and the combination of such methods may be particularly powerful. Studies applying these methods help us to understand how neuronal activity, optical imaging, and functional magnetic resonance imaging signals are related within the temporal lobe, and to uncover the fine-grained and large-scale spatial organization of object and face representations in the primate brain.

The effect of labeling parameters on perfusion-based fMRI in nonhuman primates.

The blood oxygenation level-dependent (BOLD) signal is the most commonly used modality of functional magnetic resonance imaging (fMRI) today. Although easy to implement, it is an ambiguous signal since it results from a combination of several hemodynamic factors. Functional cerebral blood flow changes, as measured by using arterial spin labeling (ASL), typically occur in the parenchyma and have been demonstrated to be more closely coupled to neural activation compared with BOLD. However, the intrinsically low signals from ASL techniques have hindered its widespread application to fMRI for basic research and even more so for clinical applications. Here, we report the first implementation of continuous ASL in the anaesthetized macaque at high magnetic field of 7 T. The technique was optimized to permit maximum signal-to-noise ratio of functional perfusion-based images at high spatial resolution. The effect of labeling parameters, such as label time and post-label delay (PLD), on functional cerebral blood flow (fCBF) in the visual cortex was evaluated. Functional cerebral blood flow maps did not change with increasing label time after 2,000 ms, indicating that a label time of 2,000 ms is sufficient for reliable mapping of fCBF. The percent changes obtained using fCBF were better localized to gray matter, than those obtained with BOLD. A short PLD of 200 ms revealed significantly higher fCBF changes at the cortical surface, indicating large-vessel contamination, than a long PLD of 800 ms. However, the effect of the PLD on fCBF was smaller than on baseline CBF. These results are of importance for high-resolution applications, and when accurate quantification is required for studies in monkeys as well as in humans.

High-resolution fMRI of macaque V1.

To understand the physiological mechanisms underlying the blood-oxygenation-level-dependent (BOLD) signal, the acquisition of data must be optimized to achieve the maximum possible spatial resolution and specificity. The term "specificity" implies the selective enhancement of signals originating in the parenchyma, and thus best reflecting actual neural activity. Such spatial specificity is a prerequisite for imaging aimed at the elucidation of interactions between cortical micromodules, such as columns and laminae. In addition to the optimal selection of functional magnetic resonance imaging pulse sequences, accurate superposition of activation patterns onto corresponding anatomical scans, preferably acquired during the same experimental session, is necessary. At high resolution, exact functional-to-structural registration is of critical importance, because even small differences in geometry, that arise when different sequences are used for functional and anatomical scans, can lead to misallocation of activation and erroneous interpretation of data. In the present study, we used spin-echo (SE) echo planar imaging (EPI) for functional scans, since the SE-BOLD signal is sensitive to the capillary response, together with SE-EPI anatomical reference scans. The combination of these acquisition methods revealed a clear spatial colocalization of the largest fractional changes with the Gennari line, suggesting peak activity in Layer IV. Notably, this very same layer coincided with the largest relaxivity changes as observed in steady-state cerebral blood volume measurements, using the intravascular agent monocrystalline iron oxide nanoparticles (MION).

Laminar specificity in monkey V1 using high-resolution SE-fMRI.

The lamination of mammalian neocortex is widely used as reference for describing a wide range of anatomical and physiological data. Its value lies in the observation that in all examined species, cortical afferents, intrinsic cells and projection neurons organize themselves with respect to the laminae. The comprehension of the computations, carried out by the neocortical microcircuits, critically relies on the study of the interlaminar connectivity patterns and the intralaminar physiological processes in vivo. High-resolution functional neuroimaging, enabling the visualization of activity in individual cortical laminae or columns, may greatly contribute in such studies. Yet, the BOLD effect, as measured with the commonly used GE-EPI, contains contributions from both macroscopic venous blood vessels and capillaries. The low density of the cortical veins limits the effective spatial specificity of the fMRI signal and yields maps that are weighted toward the macrovasculature, which thus can be significantly different from the actual site of increased neuronal activity. Spin-echo (SE) sequences yielding apparent T2-weighted BOLD images have been shown to improve spatial specificity by increasing the sensitivity of the signal to spins of the parenchyma, particularly at high magnetic fields. Here we used SE-fMRI at 4.7 T to examine the specificity and resolution of functional maps obtained by stimulating the primary visual cortex of monkeys. Cortical layers could be clearly visualized, and functional activity was predominantly localized in cortical layer IV/Duvernoy layer 3. The choice of sequence parameters influences the fMRI signal, as the SE-EPI is by nature sensitive to T2* in addition to its T2 dependency. Using parameters that limit T2* effects yielded higher specificity and better visualization of the cortical laminae. Because the demands of high-spatial resolution using SE severely decreases temporal resolution, we used a stimulus protocol that allows sampling at higher effective temporal resolution. This way, it was possible to acquire high-spatial and high-temporal resolution SE-fMRI data.

Effects of noise bandwidth and amplitude modulation on masking in frog auditory midbrain neurons.

Natural auditory scenes such as frog choruses consist of multiple sound sources (i.e., individual vocalizing males) producing sounds that overlap extensively in time and spectrum, often in the presence of other biotic and abiotic background noise. Detection of a signal in such environments is challenging, but it is facilitated when the noise shares common amplitude modulations across a wide frequency range, due to a phenomenon called comodulation masking release (CMR). Here, we examined how properties of the background noise, such as its bandwidth and amplitude modulation, influence the detection threshold of a target sound (pulsed amplitude modulated tones) by single neurons in the frog auditory midbrain. We found that for both modulated and unmodulated masking noise, masking was generally stronger with increasing bandwidth, but it was weakened for the widest bandwidths. Masking was less for modulated noise than for unmodulated noise for all bandwidths. However, responses were heterogeneous, and only for a subpopulation of neurons the detection of the probe was facilitated when the bandwidth of the modulated masker was increased beyond a certain bandwidth - such neurons might contribute to CMR. We observed evidence that suggests that the dips in the noise amplitude are exploited by TS neurons, and observed strong responses to target signals occurring during such dips. However, the interactions between the probe and masker responses were nonlinear, and other mechanisms, e.g., selective suppression of the response to the noise, may also be involved in the masking release.

fMRI of the temporal lobe of the awake monkey at 7 T.

Increasingly 7 T scanners are used for fMRI of humans and non-human primates, promising improvements in signal-to-noise, spatial resolution and specificity. A disadvantage of fMRI at 7 T, but already at 3 T, is that susceptibility artifacts from air-filled cavities like the ear canal and nasal cavity cause signal loss and distortion. This limits the applicability of fMRI in these areas, thereby limiting study of these areas, but it also limits study of processes that span large-scale cortical networks or the entire brain. Our goal is to study the inferior temporal (IT) lobe in awake monkeys because of its importance in object perception and recognition, but the functional signal is degraded by strong susceptibility gradients. To allow fMRI of this region, we used an optimized SE-EPI, which recovers signal lost with GE-EPI and we corrected for susceptibility-induced image distortion. SE-EPI has the added advantage that, in contrast to GE-EPI, where the functional signal derives to a large extent from veins, the SE-EPI signal arises from the microvasculature, and hence it better represents the neural activation. We show fMRI at 7 T of the entire visual pathway in the awake primate with robust and widespread activation in all ventral areas of the brain, including areas adjacent to the ear canal. This allows fMRI of areas that normally suffer from artifact and thus more reliable whole-brain studies.

Seasonal changes in frequency tuning and temporal processing in single neurons in the frog auditory midbrain.

Frogs rely on acoustic signaling to detect, discriminate, and localize mates. In the temperate zone, reproduction occurs in the spring, when frogs emerge from hibernation and engage in acoustically guided behaviors. In response to the species mating call, males typically show evoked vocal responses or other territorial behaviors, and females show phonotactic responses. Because of their strong seasonal behavior, it is possible that the frog auditory system also displays seasonal variation, as evidenced in their vocal control system. This hypothesis was tested in male Northern leopard frogs by evaluating the response characteristics of single neurons in the torus semicircularis (TS; a homolog of the inferior colliculus) to a synthetic mating call at different times of the year. We found that TS neurons displayed a seasonal change in frequency tuning and temporal properties. Frequency tuning shifted from a predominance of TS units sensitive to intermediate frequencies (700-1200 Hz) in the winter, to low frequencies (100-600 Hz) in the summer. In winter and early spring, most TS neurons showed poor, or weak, time locking to the envelope of the amplitude-modulated synthetic call, whereas in late spring and early summer the majority of TS neurons showed robust time-locked responses. These seasonal differences indicate that neural coding by auditory midbrain neurons in the Northern leopard frog is subject to seasonal fluctuation.