Russell W. Chan

Brain resting-state functional MRI connectivity: Morphological foundation and plasticity

Despite the immense ongoing efforts to map brain functional connections and organizations with resting-state functional MRI (rsfMRI), the mechanisms governing the temporally coherent rsfMRI signals remain unclear. In particular, there is a lack of direct evidence regarding the morphological foundation and plasticity of these rsfMRI derived connections. In this study, we investigated the role of axonal projections in rsfMRI connectivity and its plasticity. Well-controlled rodent models of complete and posterior corpus callosotomy were longitudinally examined with rsfMRI at 7T in conjunction with intracortical EEG recording and functional MRI tracing of interhemispheric neuronal pathways by manganese (Mn(2+)). At post-callosotomy day 7, significantly decreased interhemispheric rsfMRI connectivity was observed in both groups in the specific cortical areas whose callosal connections were severed. At day 28, the disrupted connectivity was restored in the partial callosotomy group but not in the complete callosotomy group, likely due to the compensation that occurred through the remaining interhemispheric axonal pathways. This restoration - along with the increased intrahemispheric functional connectivity observed in both groups at day 28 - highlights the remarkable adaptation and plasticity in brain rsfMRI connections. These rsfMRI findings were paralleled by the intracortical EEG recording and Mn(2+) tracing results. Taken together, our experimental results directly demonstrate that axonal connections are the indispensable foundation for rsfMRI connectivity and that such functional connectivity can be plastic and dynamically reorganized atop the morphological connections.

In vivo visuotopic brain mapping with manganese-enhanced MRI and resting-state functional connectivity MRI

The rodents are an increasingly important model for understanding the mechanisms of development, plasticity, functional specialization and disease in the visual system. However, limited tools have been available for assessing the structural and functional connectivity of the visual brain network globally, in vivo and longitudinally. There are also ongoing debates on whether functional brain connectivity directly reflects structural brain connectivity. In this study, we explored the feasibility of manganese-enhanced MRI (MEMRI) via 3 different routes of Mn(2+) administration for visuotopic brain mapping and understanding of physiological transport in normal and visually deprived adult rats. In addition, resting-state functional connectivity MRI (RSfcMRI) was performed to evaluate the intrinsic functional network and structural-functional relationships in the corresponding anatomical visual brain connections traced by MEMRI. Upon intravitreal, subcortical, and intracortical Mn(2+) injection, different topographic and layer-specific Mn enhancement patterns could be revealed in the visual cortex and subcortical visual nuclei along retinal, callosal, cortico-subcortical, transsynaptic and intracortical horizontal connections. Loss of visual input upon monocular enucleation to adult rats appeared to reduce interhemispheric polysynaptic Mn(2+) transfer but not intra- or inter-hemispheric monosynaptic Mn(2+) transport after Mn(2+) injection into visual cortex. In normal adults, both structural and functional connectivity by MEMRI and RSfcMRI was stronger interhemispherically between bilateral primary/secondary visual cortex (V1/V2) transition zones (TZ) than between V1/V2 TZ and other cortical nuclei. Intrahemispherically, structural and functional connectivity was stronger between visual cortex and subcortical visual nuclei than between visual cortex and other subcortical nuclei. The current results demonstrated the sensitivity of MEMRI and RSfcMRI for assessing the neuroarchitecture, neurophysiology and structural-functional relationships of the visual brains in vivo. These may possess great potentials for effective monitoring and understanding of the basic anatomical and functional connections in the visual system during development, plasticity, disease, pharmacological interventions and genetic modifications in future studies.

Low-frequency hippocampal–cortical activity drives brain-wide resting-state functional MRI connectivity

Significance The hippocampus with its dense reciprocal axonal projections to and from cortex is widely believed to mediate numerous cognitive functions. However, it is unknown whether and how specific hippocampal–cortical activity contributes to the brain-wide functional connectivity. Here, we use optogenetics and fMRI to examine how excitatory neural activity initiated in the dorsal dentate gyrus of the hippocampus propagates and modulates resting-state fMRI (rsfMRI) connectivity. We discover its robust propagation brain-wide at low frequency (1 Hz), which enhances interhemispheric rsfMRI connectivity and cortical and subcortical visual responses. Our findings highlight the important role of slow hippocampal–cortical oscillatory activity in driving brain-wide rsfMRI connectivity and mediating sensory processing. The hippocampus, including the dorsal dentate gyrus (dDG), and cortex engage in bidirectional communication. We propose that low-frequency activity in hippocampal–cortical pathways contributes to brain-wide resting-state connectivity to integrate sensory information. Using optogenetic stimulation and brain-wide fMRI and resting-state fMRI (rsfMRI), we determined the large-scale effects of spatiotemporal-specific downstream propagation of hippocampal activity. Low-frequency (1 Hz), but not high-frequency (40 Hz), stimulation of dDG excitatory neurons evoked robust cortical and subcortical brain-wide fMRI responses. More importantly, it enhanced interhemispheric rsfMRI connectivity in various cortices and hippocampus. Subsequent local field potential recordings revealed an increase in slow oscillations in dorsal hippocampus and visual cortex, interhemispheric visual cortical connectivity, and hippocampal–cortical connectivity. Meanwhile, pharmacological inactivation of dDG neurons decreased interhemispheric rsfMRI connectivity. Functionally, visually evoked fMRI responses in visual regions also increased during and after low-frequency dDG stimulation. Together, our results indicate that low-frequency activity robustly propagates in the dorsal hippocampal–cortical pathway, drives interhemispheric cortical rsfMRI connectivity, and mediates visual processing.

Long-range projections coordinate distributed brain-wide neural activity with a specific spatiotemporal profile

Significance What makes the brain tick? A simple yet challenging question that has captivated our minds for centuries. This sentiment was fittingly reflected in the launch of The BRAIN Initiative 3 years ago, spurred by the rapid advancement of noninvasive brain imaging and neuronal mapping technologies that have advanced our understanding of neural networks, which are central to brain functions and behavior. Here, we study the patterns of large-scale brain-wide interactions mediated by thalamo-cortical networks through optogenetics and functional MRI. We found that the thalamus can recruit long-range cortical and subcortical networks and initiate their interactions in a spatiotemporally specific manner. This finding provides a fresh impetus to study the mysteries of the brain. One challenge in contemporary neuroscience is to achieve an integrated understanding of the large-scale brain-wide interactions, particularly the spatiotemporal patterns of neural activity that give rise to functions and behavior. At present, little is known about the spatiotemporal properties of long-range neuronal networks. We examined brain-wide neural activity patterns elicited by stimulating ventral posteromedial (VPM) thalamo-cortical excitatory neurons through combined optogenetic stimulation and functional MRI (fMRI). We detected robust optogenetically evoked fMRI activation bilaterally in primary visual, somatosensory, and auditory cortices at low (1 Hz) but not high frequencies (5–40 Hz). Subsequent electrophysiological recordings indicated interactions over long temporal windows across thalamo-cortical, cortico-cortical, and interhemispheric callosal projections at low frequencies. We further observed enhanced visually evoked fMRI activation during and after VPM stimulation in the superior colliculus, indicating that visual processing was subcortically modulated by low-frequency activity originating from VPM. Stimulating posteromedial complex thalamo-cortical excitatory neurons also evoked brain-wide blood-oxygenation-level–dependent activation, although with a distinct spatiotemporal profile. Our results directly demonstrate that low-frequency activity governs large-scale, brain-wide connectivity and interactions through long-range excitatory projections to coordinate the functional integration of remote brain regions. This low-frequency phenomenon contributes to the neural basis of long-range functional connectivity as measured by resting-state fMRI.

BOLD fMRI study of ultrahigh frequency encoding in the inferior colliculus

Many vertebrates communicate with ultrahigh frequency (UHF) vocalizations to limit auditory detection by predators. The mechanisms underlying the neural encoding of such UHF sounds may provide important insights for understanding neural processing of other complex sounds (e.g. human speeches). In the auditory system, sound frequency is normally encoded topographically as tonotopy, which, however, contains very limited representation of UHFs in many species. Instead, electrophysiological studies suggested that two neural mechanisms, both exploiting the interactions between frequencies, may contribute to UHF processing. Neurons can exhibit excitatory or inhibitory responses to a tone when another UHF tone is presented simultaneously (combination sensitivity). They can also respond to such stimulation if they are tuned to the frequency of the cochlear-generated distortion products of the two tones, e.g. their difference frequency (cochlear distortion). Both mechanisms are present in an early station of the auditory pathway, the midbrain inferior colliculus (IC). Currently, it is unclear how prevalent the two mechanisms are and how they are functionally integrated in encoding UHFs. This study investigated these issues with large-view BOLD fMRI in rat auditory system, particularly the IC. UHF vocalizations (above 40kHz), but not pure tones at similar frequencies (45, 55, 65, 75kHz), evoked robust BOLD responses in multiple auditory nuclei, including the IC, reinforcing the sensitivity of the auditory system to UHFs despite limited representation in tonotopy. Furthermore, BOLD responses were detected in the IC when a pair of UHF pure tones was presented simultaneously (45 & 55kHz, 55 & 65kHz, 45 & 65kHz, 45 & 75kHz). For all four pairs, a cluster of voxels in the ventromedial side always showed the strongest responses, displaying combination sensitivity. Meanwhile, voxels in the dorsolateral side that showed strongest secondary responses to each pair of UHF pure tones also showed the strongest responses to a pure tone at their difference frequency, suggesting that they are sensitive to cochlear distortion. These BOLD fMRI results indicated that combination sensitivity and cochlear distortion are employed by large but spatially distinctive neuron populations in the IC to represent UHFs. Our imaging findings provided insights for understanding sound feature encoding in the early stage of the auditory pathway.

Structural and Functional Brain Remodeling during Pregnancy with Diffusion Tensor MRI and Resting-State Functional MRI

Although pregnancy-induced hormonal changes have been shown to alter the brain at the neuronal level, the exact effects of pregnancy on brain at the tissue level remain unclear. In this study, diffusion tensor imaging (DTI) and resting-state functional MRI (rsfMRI) were employed to investigate and document the effects of pregnancy on the structure and function of the brain tissues. Fifteen Sprague-Dawley female rats were longitudinally studied at three days before mating (baseline) and seventeen days after mating (G17). G17 is equivalent to the early stage of the third trimester in humans. Seven age-matched nulliparous female rats served as non-pregnant controls and were scanned at the same time-points. For DTI, diffusivity was found to generally increase in the whole brain during pregnancy, indicating structural changes at microscopic levels that facilitated water molecular movement. Regionally, mean diffusivity increased more pronouncedly in the dorsal hippocampus while fractional anisotropy in the dorsal dentate gyrus increased significantly during pregnancy. For rsfMRI, bilateral functional connectivity in the hippocampus increased significantly during pregnancy. Moreover, fractional anisotropy increase in the dentate gyrus appeared to correlate with the bilateral functional connectivity increase in the hippocampus. These findings revealed tissue structural modifications in the whole brain during pregnancy, and that the hippocampus was structurally and functionally remodeled in a more marked manner.

Longitudinal metabolic changes in the hippocampus and thalamus of the maternal brain revealed by proton magnetic resonance spectroscopy

Pregnancy is accompanied by dramatic hormonal changes, which are essential for the display of maternal behaviors. Reproductive hormones have been shown to remodel the neuronal structure and function of the female brain. However, most previous studies have examined the structural and functional changes elicited by transient fluctuations in reproductive hormones. The impact of naturally elevated and more sustained hormonal alterations during pregnancy and lactation are not fully understood. Further alterations in neurochemistry, which may result in substantial changes in the structure and function of neurons that are associated with behavioral modifications in the maternal female, are difficult to capture in a longitudinal and non-invasive manner. In this study, neurobiological alterations during pregnancy and motherhood were investigated longitudinally using non-invasive proton magnetic resonance spectroscopy ((1)H MRS) at 7T in regions related to learning and memory, such as the hippocampus, and in structures involved in alertness and attention, such as the thalamus. Pregnant primiparous rats (N=15) were studied at three days before mating, gestational day 17, lactation day 7 and post-weaning day 7. Age-matched nulliparous female rats (N=9) served as non-pregnant controls. Significantly higher N-acetylaspartate (NAA) levels were observed in the hippocampus and thalamus of rats at gestational day 17. These increases may be associated with increased dendritic sprouting, synaptogenesis or neurogenesis, thereby facilitating supporting behaviors that involve spatial learning and memory and alleviating fear and stress. The (1)H MRS detection of ongoing neurochemical changes induced by pregnancy, especially in the hippocampus, can shed light on the neurochemical underpinnings of behavioral modifications, including the improvement in spatial learning and memory, during pregnancy.

Resting-State fMRI Using Passband Balanced Steady- State Free Precession

Objective Resting-state functional MRI (rsfMRI) has been increasingly used for understanding brain functional architecture. To date, most rsfMRI studies have exploited blood oxygenation level-dependent (BOLD) contrast using gradient-echo (GE) echo planar imaging (EPI), which can suffer from image distortion and signal dropout due to magnetic susceptibility and inherent long echo time. In this study, the feasibility of passband balanced steady-state free precession (bSSFP) imaging for distortion-free and high-resolution rsfMRI was investigated. Methods rsfMRI was performed in humans at 3 T and in rats at 7 T using bSSFP with short repetition time (TR = 4/2.5 ms respectively) in comparison with conventional GE-EPI. Resting-state networks (RSNs) were detected using independent component analysis. Results and Significance RSNs derived from bSSFP images were shown to be spatially and spectrally comparable to those derived from GE-EPI images with considerable intra- and inter-subject reproducibility. High-resolution bSSFP images corresponded well to the anatomical images, with RSNs exquisitely co-localized to the gray matter. Furthermore, RSNs at areas of severe susceptibility such as human anterior prefrontal cortex and rat piriform cortex were proved accessible. These findings demonstrated for the first time that passband bSSFP approach can be a promising alternative to GE-EPI for rsfMRI. It offers distortion-free and high-resolution RSNs and is potentially suited for high field studies.

Optogenetic auditory fMRI reveals the effects of visual cortical inputs on auditory midbrain response

Sensory cortices contain extensive descending (corticofugal) pathways, yet their impact on brainstem processing – particularly across sensory systems – remains poorly understood. In the auditory system, the inferior colliculus (IC) in the midbrain receives cross-modal inputs from the visual cortex (VC). However, the influences from VC on auditory midbrain processing are unclear. To investigate whether and how visual cortical inputs affect IC auditory responses, the present study combines auditory blood-oxygenation-level-dependent (BOLD) functional MRI (fMRI) with cell-type specific optogenetic manipulation of visual cortex. The results show that predominant optogenetic excitation of the excitatory pyramidal neurons in the infragranular layers of the primary VC enhances the noise-evoked BOLD fMRI responses within the IC. This finding reveals that inputs from VC influence and facilitate basic sound processing in the auditory midbrain. Such combined optogenetic and auditory fMRI approach can shed light on the large-scale modulatory effects of corticofugal pathways and guide detailed electrophysiological studies in the future.