Kentaro Miyamoto

Functional Connectivity between Anatomically Unconnected Areas Is Shaped by Collective Network-Level Effects in the Macaque Cortex

Coherent spontaneous blood oxygen level-dependent (BOLD) fluctuations have been intensely investigated as a measure of functional connectivity (FC) in the primate neocortex. BOLD-FC is commonly assumed to be constrained by the underlying anatomical connectivity (AC); however, cortical area pairs with no direct AC can also have strong BOLD-FC. On the mechanism generating FC in the absence of direct AC, there are 2 possibilities: 1) FC is determined by signal flows via short connection patterns, such as serial relays and common afferents mediated by a third area; 2) FC is shaped by collective effects governed by network properties of the cortex. In this study, we conducted functional magnetic resonance imaging in anesthetized macaque monkeys and found that BOLD-FC between unconnected areas depends less on serial relays through a third area than on common afferents and, unexpectedly, common efferents, which does not match the first possibility. By utilizing a computational model for interareal BOLD-FC network, we show that the empirically detected AC-FC relationships reflect the configuration of network building blocks (motifs) in the cortical anatomical network, which supports the second possibility. Our findings indicate that FC is not determined solely by interareal short connection patterns but instead is substantially influenced by the network-level cortical architecture.

Functional differentiation of memory retrieval network in macaque posterior parietal cortex.

Human fMRI studies revealed involvement of the posterior parietal cortex (PPC) during memory retrieval. However, corresponding memory-related regions in macaque PPC have not been established. In this monkey fMRI study, comparisons of cortical activity during correct recognition of previously seen items and rejection of unseen items revealed two major PPC activation sites that were differentially characterized by a serial probe recognition paradigm: area PG/PGOp in inferior parietal lobule, along with the hippocampus, was more active for initial item retrieval, while area PEa/DIP in intraparietal sulcus was for the last item. Effective connectivity analyses revealed that connectivity from hippocampus to PG/PGOp, but not to PEa/DIP, increased during initial item retrieval. The two parietal areas with differential serial probe recognition profiles were embedded in two different subnetworks of the brain-wide retrieval-related regions. These functional dissociations in the macaque PPC imply the functional correspondence of retrieval-related PPC networks in macaques and humans.

Dissociable Memory Traces within the Macaque Medial Temporal Lobe Predict Subsequent Recognition Performance

Functional magnetic resonance imaging (fMRI) studies have revealed that activity in the medial temporal lobe (MTL) predicts subsequent memory performance in humans. Because of limited knowledge on cytoarchitecture and axonal projections of the human MTL, precise localization and characterization of the areas that can predict subsequent memory performance are benefited by the use of nonhuman primates in which integrated approach of the MRI- and cytoarchiture-based boundary delineation is available. However, neural correlates of this subsequent memory effect have not yet been identified in monkeys. Here, we used fMRI to examine activity in the MTL during memory encoding of events that monkeys later remembered or forgot. Application of both multivoxel pattern analysis and conventional univariate analysis to high-resolution fMRI data allowed us to identify memory traces within the caudal entorhinal cortex (cERC) and perirhinal cortex (PRC), as well as within the hippocampus proper. Furthermore, activity in the cERC and the hippocampus, which are directly connected, was responsible for encoding the initial items of sequentially presented pictures, which may reflect recollection-like recognition, whereas activity in the PRC was not. These results suggest that two qualitatively distinct encoding processes work in the monkey MTL and that recollection-based memory is formed by the interplay of the hippocampus with the cERC, a focal cortical area anatomically closer to the hippocampus and hierarchically higher than previously believed. These findings will advance the understanding of common memory system between humans and monkeys and accelerate fine electrophysiological characterization of these dissociable memory traces in the monkey MTL.

Causal neural network of metamemory for retrospection in primates

Are you aware how well you remember? Self-monitoring and evaluation of our own memory is a mental process called metamemory. For metamemory, we need access to information about the strength of our own memory traces. The brain structures and neural mechanisms involved in metamemory are completely unknown. Miyamoto et al. devised a test paradigm for metamemory in macaques, in which the monkeys judged their own confidence in remembering past experiences. The authors combined this approach with functional brain imaging to reveal the neural substrates of metamemory for retrospection. A specific region in the prefrontal brain was essential for meta mnemonic decision-making. Inactivation of this region caused selective impairment of metamemory, but not of memory itself. Science, this issue p. 188 Macaques possess brain areas for retrospection whose inactivation causes selective impairment in metamemory. We know how confidently we know: Metacognitive self-monitoring of memory states, so-called “metamemory,” enables strategic and efficient information collection based on past experiences. However, it is unknown how metamemory is implemented in the brain. We explored causal neural mechanism of metamemory in macaque monkeys performing metacognitive confidence judgments on memory. By whole-brain searches via functional magnetic resonance imaging, we discovered a neural correlate of metamemory for temporally remote events in prefrontal area 9 (or 9/46d), along with that for recent events within area 6. Reversible inactivation of each of these identified loci induced doubly dissociated selective impairments in metacognitive judgment performance on remote or recent memory, without impairing recognition performance itself. The findings reveal that parallel metamemory streams supervise recognition networks for remote and recent memory, without contributing to recognition itself.

fMRI Activity in the Macaque Cerebellum Evoked by Intracortical Microstimulation of the Primary Somatosensory Cortex: Evidence for Polysynaptic Propagation

Simultaneous electrical microstimulation (EM) and functional magnetic resonance imaging (fMRI) is a useful tool for probing connectivity across brain areas in vivo. However, it is not clear whether intracortical EM can evoke blood-oxygenation-level-dependent (BOLD) signal in areas connected polysynaptically to the stimulated site. To test for the presence of the BOLD activity evoked by polysynaptic propagation of the EM signal, we conducted simultaneous fMRI and EM in the primary somatosensory cortex (S1) of macaque monkeys. We in fact observed BOLD activations in the contralateral cerebellum which is connected to the stimulation site (i.e. S1) only through polysynaptic pathways. Furthermore, the magnitude of cerebellar activations was dependent on the current amplitude of the EM, confirming the EM is the cause of the cerebellar activations. These results suggest the importance of considering polysynaptic signal propagation, particularly via pathways including subcortical structures, for correctly interpreting ‘functional connectivity’ as assessed by simultaneous EM and fMRI.

Dynamically allocated hub in task-evoked network predicts the vulnerable prefrontal locus for contextual memory retrieval in macaques

Neuroimaging and neurophysiology have revealed that multiple areas in the prefrontal cortex (PFC) are activated in a specific memory task, but severity of impairment after PFC lesions is largely different depending on which activated area is damaged. The critical relationship between lesion sites and impairments has not yet been given a clear mechanistic explanation. Although recent works proposed that a whole-brain network contains hubs that play integrative roles in cortical information processing, this framework relying on an anatomy-based structural network cannot account for the vulnerable locus for a specific task, lesioning of which would bring impairment. Here, we hypothesized that (i) activated PFC areas dynamically form an ordered network centered at a task-specific “functional hub” and (ii) the lesion-effective site corresponds to the “functional hub,” but not to a task-invariant “structural hub.” To test these hypotheses, we conducted functional magnetic resonance imaging experiments in macaques performing a temporal contextual memory task. We found that the activated areas formed a hierarchical hub-centric network based on task-evoked directed connectivity, differently from the anatomical network reflecting axonal projection patterns. Using a novel simulated-lesion method based on support vector machine, we estimated severity of impairment after lesioning of each area, which accorded well with a known dissociation in contextual memory impairment in macaques (impairment after lesioning in area 9/46d, but not in area 8Ad). The predicted severity of impairment was proportional to the network “hubness” of the virtually lesioned area in the task-evoked directed connectivity network, rather than in the anatomical network known from tracer studies. Our results suggest that PFC areas dynamically and cooperatively shape a functional hub-centric network to reallocate the lesion-effective site depending on the cognitive processes, apart from static anatomical hubs. These findings will be a foundation for precise prediction of behavioral impacts of damage or surgical intervention in human brains.

Conversion of object identity to object-general semantic value in the primate temporal cortex

Faulty remembrance of objects past The primate brain analyzes visual input along the ventral processing stream to extract the identity of an object. The final stage of this stream, the perirhinal cortex, plays a crucial role in object recognition. Tamura et al. systematically biased the judgments of monkeys in an old-new object recognition task by using either optogenetic or electrical stimulation. The monkeys judged an encountered object as familiar when the stimulation site was in a hotspot where memory neurons were clustered. However, at the hotspots fringe region, where neurons lost selective responses to the learned objects, electrical microstimulation led the monkeys to mistakenly judge an object as never seen before. Science, this issue p. 687 Optogenetic activation of perirhinal memory neurons in primates drives a subjective feeling of having previously seen an object. At the final stage of the ventral visual stream, perirhinal neurons encode the identity of memorized objects through learning. However, it remains elusive whether and how object percepts alone, or concomitantly a nonphysical attribute of the objects (“learned”), are decoded from perirhinal activities. By combining monkey psychophysics with optogenetic and electrical stimulations, we found a focal spot of memory neurons where both stimulations led monkeys to preferentially judge presented objects as “already seen.” In an adjacent fringe area, where neurons did not exhibit selective responses to the learned objects, electrical stimulation induced the opposite behavioral bias toward “never seen before,” whereas optogenetic stimulation still induced bias toward “already seen.” These results suggest that mnemonic judgment of objects emerges via the decoding of their nonphysical attributes encoded by perirhinal neurons.

Remapping of memory encoding and retrieval networks: insights from neuroimaging in primates.

Advancements in neuroimaging techniques have allowed for the investigation of the neural correlates of memory functions in the whole human brain. Thus, the involvement of various cortical regions, including the medial temporal lobe (MTL) and posterior parietal cortex (PPC), has been repeatedly reported in the human memory processes of encoding and retrieval. However, the functional roles of these sites could be more fully characterized utilizing nonhuman primate models, which afford the potential for well-controlled, finer-scale experimental procedures that are inapplicable to humans, including electrophysiology, histology, genetics, and lesion approaches. Yet, the presence and localization of the functional counterparts of these human memory-related sites in the macaque monkey MTL or PPC were previously unknown. Therefore, to bridge the inter-species gap, experiments were required in monkeys using functional magnetic resonance imaging (fMRI), the same methodology adopted in human studies. Here, we briefly review the history of experimentation on memory systems using a nonhuman primate model and our recent fMRI studies examining memory processing in monkeys performing recognition memory tasks. We will discuss the memory systems common to monkeys and humans and future directions of finer cell-level characterization of memory-related processes using electrophysiological recording and genetic manipulation approaches.

Molecular hydrogen suppresses activated Wnt/β-catenin signaling

Molecular hydrogen (H2) is effective for many diseases. However, molecular bases of H2 have not been fully elucidated. Cumulative evidence indicates that H2 acts as a gaseous signal modulator. We found that H2 suppresses activated Wnt/β-catenin signaling by promoting phosphorylation and degradation οf β-catenin. Either complete inhibition of GSK3 or mutations at CK1- and GSK3-phosphorylation sites of β-catenin abolished the suppressive effect of H2. H2 did not increase GSK3-mediated phosphorylation of glycogen synthase, indicating that H2 has no direct effect on GSK3 itself. Knock-down of adenomatous polyposis coli (APC) or Axin1, which form the β-catenin degradation complex, minimized the suppressive effect of H2 on β-catenin accumulation. Accordingly, the effect of H2 requires CK1/GSK3-phosphorylation sites of β-catenin, as well as the β-catenin degradation complex comprised of CK1, GSK3, APC, and Axin1. We additionally found that H2 reduces the activation of Wnt/β-catenin signaling in human osteoarthritis chondrocytes. Oral intake of H2 water tended to ameliorate cartilage degradation in a surgery-induced rat osteoarthritis model through attenuating β-catenin accumulation. We first demonstrate that H2 suppresses abnormally activated Wnt/β-catenin signaling, which accounts for the protective roles of H2 in a fraction of diseases.

Fluoxetine ameliorates cartilage degradation in osteoarthritis by inhibiting Wnt/β-catenin signaling

Abnormal activation of the Wnt/β-catenin signaling is implicated in the osteoarthritis (OA) pathology. We searched for a pre-approved drug that suppresses abnormally activated Wnt/β-catenin signaling and has a potency to reduce joint pathology in OA. We introduced the TOPFlash reporter plasmid into HCS-2/8 human chondrosarcoma cells to estimate the Wnt/β-catenin activity in the presence of 10 μM each compound in a panel of pre-approved drugs. We found that fluoxetine, an antidepressant in the class of selective serotonin reuptake inhibitors (SSRI), down-regulated Wnt/β-catenin signaling in human chondrosarcoma cells. Fluoxetine inhibited both Wnt3A- and LiCl-induced loss of proteoglycans in chondrogenically differentiated ATDC5 cells. Fluoxetine increased expression of Sox9 (the chondrogenic master regulator), and decreased expressions of Axin2 (a marker for Wnt/β-catenin signaling) and Mmp13 (matrix metalloproteinase 13). Fluoxetine suppressed a LiCl-induced increase of total β-catenin and a LiCl-induced decrease of phosphorylated β-catenin in a dose-dependent manner. An in vitro protein-binding assay showed that fluoxetine enhanced binding of β-catenin with Axin1, which is a scaffold protein forming the degradation complex for β-catenin. Fluoxetine suppressed LiCl-induced β-catenin accumulation in human OA chondrocytes. Intraarticular injection of fluoxetine in a rat OA model ameliorated OA progression and suppressed β-catenin accumulation.