Structural connectivity of the precuneus and its relation to resting-state networks.

The precuneus is an association area in the posteromedial cortex (PMC) that is involved in high-order cognitive functions through integrating multi-modal information. Previous studies have shown that the precuneus is functionally heterogeneous and subdivided into several subfields organized by the anterior-posterior and ventral-dorsal axes. Further, the precuneus forms the structural core of brain connectivity as a rich-club hub and overlaps with the default mode network (DMN) as the functional core. This review summarizes recent research on the connectivity and cognitive functions of the precuneus. We then present our recent tractography-based studies of the precuneus and contextual these results here with respect to possible cognitive functions and resting-state networks.

Spatiotemporal expression patterns of ZBP1 in the brain of mouse experimental stroke model.

Z-DNA binding protein 1 (ZBP1) is a cytosolic nucleic acid sensor, functioning as a critical mediator of inflammation and cell death pathways. Since neuroinflammation could occur in response to damage-associated molecular patterns (DAMPs), ZBP1 might be involved in neuroinflammation after stroke. However, the spatiotemporal expression profile of ZBP1 in the post-stroke brain remains to be elucidated. The aim of this study is to demonstrate the spatiotemporal expression patterns of ZBP1 in the post-stroke brain using a mouse photothrombotic stroke model. Real-time PCR assays showed that ZBP1 is induced on days 3-14 post stroke. ZBP1 immunoreactivity was observed in Iba1-positive microglia/macrophages in peri-infarct regions by immunohistochemistry. ZBP1-positive cells were spread in layers surrounding the infarct core by 7-14 days post stroke. Interestingly, ZBP1 immunoreactivity was also detected in CD206-positive border-associated macrophages (BAMs) in the meninges. Furthermore, ZBP1-expressing cells were positive for antibodies against inflammatory mediators such as Toll-like receptor 4 (TLR4), Toll/IL-1R domain-containing adaptor-inducing IFN-ß (TRIF), and receptor-interacting serine/threonine-protein kinase 1 (RIPK1). Morphological analysis with confocal microscopy showed that the co-localization signals of ZBP1 and its adaptor, TRIF, are increased by glucose oxidase (GOx) treatment, which has been reported to induce mitochondrial DNA (mtDNA) release. These results suggest that ZBP1 is induced in peri-infarct microglia/macrophages and may be involved in DAMPs-mediated neuroinflammation involving mtDNA in the post-infarct brain.

Characteristic cortico-cortical connection profile of human precuneus revealed by probabilistic tractography.

It is generally hypothesized that functional connectivity (FC) reflects the underlying structural connectivity (SC). The precuneus is associated with highly integrated cognitive functions. However, our understanding of the structural connections that could underlie them is limited. This study aimed to characterize the cortico-cortical connections by probabilistic tractography. The precuneus corresponds to the five cortical areas (7Am, PCV, 7Pm, 7m, POS2) on the HCP MMP atlas. We first conducted the atlas-based probabilistic tractography. The anterior part (7Am) was strongly connected to the sensorimotor region. The dorsal part (7Am, 7Pm) was highly connected with the adjacent parietal and temporal cortex, while the ventral part (PCV, 7m) showed strong connections with the adjacent posterior cingulate and medial prefrontal cortex. The most posterior part (POS2) was explicitly connected to the visual cortex. In addition, there was a correlation between SC and resting-state fMRI connectivity (Spearman's rank correlation coefficient = 0.322 ± 0.019, p < 0.05 corrected at subject level). Collectively, the current study revealed the characteristic connectional profile of precuneus, which could shed light on the structural heterogeneity for the future functional analyses.

Searching for optimal machine learning model to classify mild cognitive impairment (MCI) subtypes using multimodal MRI data.

The intervention at the stage of mild cognitive impairment (MCI) is promising for preventing Alzheimer's disease (AD). This study aims to search for the optimal machine learning (ML) model to classify early and late MCI (EMCI and LMCI) subtypes using multimodal MRI data. First, the tract-based spatial statistics (TBSS) analyses showed LMCI-related white matter changes in the Corpus Callosum. The ROI-based tractography addressed the connected cortical areas by affected callosal fibers. We then prepared two feature subsets for ML by measuring resting-state functional connectivity (TBSS-RSFC method) and graph theory metrics (TBSS-Graph method) in these cortical areas, respectively. We also prepared feature subsets of diffusion parameters in the regions of LMCI-related white matter alterations detected by TBSS analyses. Using these feature subsets, we trained and tested multiple ML models for EMCI/LMCI classification with cross-validation. Our results showed the ensemble ML model (AdaBoost) with feature subset of diffusion parameters achieved better performance of mean accuracy 70%. The useful brain regions for classification were those, including frontal, parietal lobe, Corpus Callosum, cingulate regions, insula, and thalamus regions. Our findings indicated the optimal ML model using diffusion parameters might be effective to distinguish LMCI from EMCI subjects at the prodromal stage of AD.

Posterior Precuneus is Highly Connected to Medial Temporal Lobe Revealed by Tractography and White Matter Dissection.

The precuneus, involved in various cognitive processes, is considered to form the midline core of the default mode network (DMN), while the medial temporal lobe (MTL) is a subsystem of the DMN. Until now, the anatomical study of the precuneus-MTL connection is limited in humans. One possible reason is the precuneus' territory of the posteromedial cortex (PMC) is inconsistent across studies. The primary purpose of this study is to investigate the structural connectivity (SC) of precuneus-MTL, focusing on its anatomical organization using the Human Connectome Project Multi-modal Parcellation (HCP MMP) atlas. We first conducted the quantitative tractography analyses using the HCP dataset. The major streamlines originated from the posterior precuneus and were projected to the MTL extensively. Next, to complement the tractography data, we conducted the white matter dissection in the post-mortem human brain. We observed the major fiber bundles arise from the posterior precuneus extending to the anterior parahippocampal gyrus, which could support our tractography results. Then we analyzed the relationship between SC and resting-state functional connectivity (rsFC) of the precuneus-MTL. Although the SC-rsFC correlation was scarce on the whole, the posterior precuneus (POS2, 7Pm, 7m) showed a relatively high correlation (r = 0.38349, p < 0.05) with the posterior MTL (PreS, H, ProS, PHA1, PHA2). Our findings suggest the posterior precuneus is highly connected to MTL structurally, which could have an effect on the resting-state functional connectivity. In addition, the precuneus might consist of the heterogeneous connectivity-based subdivisions.

Gene expression profiling of the spinal cord at the chronic pain phase identified CDKL5 as a candidate gene for neural remodeling.

BACKGROUND: Chronic pain is a highly refractory and complicated condition that persists even without nociception. Several genome-wide gene expression analyses have shown that the immune response and inflammatory cytokines affect chronic pain establishment in the acute pain phase. However, compared with the acute phase, the chronic phase has a poorly elucidated gene expression profile. This study aimed to determine the gene expression profile in the spinal cord of a neuropathic pain mouse model in the chronic phase to elucidate the chronic pain characteristics. METHODS: We established a sciatic nerve cuff mouse model as a neuropathic pain model by placing a 2-mm section of a split PE-20 polyethylene tube around the sciatic nerve. The spinal cord was harvested at the L4-6 level at 28 postoperative days. Next, we examined differentially expressed genes (DEGs) through RNA sequencing (RNA-seq) compared with the sham group; moreover, we conducted enrichment analyses of the expressed genes. To reveal the chronic pain characteristics, we compared the gene expression profiles of the spinal cord between the acute and chronic phases in the neuropathic pain model. Among the chronic pain-related genes categorized in the dendrites, we focused on cyclin-dependent kinase-like 5 (CDKL5). We analyzed CDKL5 expression and function using real-time polymerase chain reaction (PCR), immunohistochemistry, and neurite extension assay in Neuro 2a (N2a) cells. We used three types of CDKL5 plasmids: wild type, nuclear localization signal-attached, and K42R kinase-dead CDKL5. RESULTS: We identified 403 DEGs, including 104 upregulated and 43 downregulated genes (false discovery rate < 0.01). Rather than inflammation or immune response, the most enriched terms in the chronic phase were "regulation of plasma membrane-bounded cell projection organization" and "dendrite." Real-time PCR assay confirmed increased CDKL5 expression in the ipsilateral dorsal horn. CDKL5 was broadly expressed in the ipsilateral dorsal horn across all layers. The neurite extension assay revealed that the cytoplasmic kinase function of CDKL5 was necessary for neurite outgrowth in N2a cells. CONCLUSION: RNA-seq of the spinal cord revealed that the most enriched genes during the chronic pain phase were involved in regulating axon and dendrite morphogenesis, including CDKL5. Our findings suggest that neural remodeling affects chronic pain establishment. Since patients with CDKL5 mutations have shown reduced pain perception, our findings suggest that CDKL5 in the spinal cord could result in neural remodeling during the chronic pain phase through cytoplasmic kinase activity.

Identification of a distinct association fiber tract "IPS-FG" to connect the intraparietal sulcus areas and fusiform gyrus by white matter dissection and tractography.

The intraparietal sulcus (IPS) in the posterior parietal cortex (PPC) is well-known as an interface for sensorimotor integration in visually guided actions. However, our understanding of the human neural network between the IPS and the cortical visual areas has been devoid of anatomical specificity. We here identified a distinctive association fiber tract "IPS-FG" to connect the IPS areas and the fusiform gyrus (FG), a high-level visual region, by white matter dissection and tractography. The major fiber bundles of this tract appeared to arise from the medial bank of IPS, in the superior parietal lobule (SPL), and project to the FG on the ventral temporal cortex (VTC) in post-mortem brains. This tract courses vertically at the temporo-parieto-occipital (TPO) junction where several fiber tracts intersect to connect the dorsal-to-ventral cortical regions, including the vertical occipital fasciculus (VOF). We then analyzed the structural connectivity of this tract with diffusion-MRI (magnetic resonance imaging) tractography. The quantitative tractography analysis revealed the major streamlines of IPS-FG interconnect the posterior IPS areas (e.g., IP1, IPS1) with FG (e.g., TF, FFC, VVC, PHA2, PIT) on the Human Connectome Project multimodal parcellation atlas (HCP MMP 1.0). Since the fronto-parietal network, including the posterior IPS areas, is recruited by multiple cognitive demands, the IPS-FG could play a role in the visuomotor integration as well as the top-down modulation of various cognitive functions reciprocally.

Temporal expression profiling of DAMPs-related genes revealed the biphasic post-ischemic inflammation in the experimental stroke model.

The neuroinflammation in the ischemic brain could occur as sterile inflammation in response to damage-associated molecular patterns (DAMPs). However, its long-term dynamic transcriptional changes remain poorly understood. It is also unknown whether this neuroinflammation contributes to the recovery or just deteriorates the outcome. The purpose of this study is to characterize the temporal transcriptional changes in the post-stroke brain focusing on DAMPs-related genes by RNA-sequencing during the period of 28 days. We conducted the RNA-sequencing on day 1, 3, 7, 14, 28 post-stroke in the mouse photothrombosis model. The gross morphological observation showed the ischemic lesion on the ipsilateral cortex turned into a scar with the clearance of cellular debris by day 28. The transcriptome analyses indicated that post-stroke period of 28 days was classified into four categories (I Baseline, II Acute, III Sub-acute-#1, IV Sub-acute-#2 phase). During this period, the well-known genes for DAMPs, receptors, downstream cascades, pro-inflammatory cytokines, and phagocytosis were transcriptionally increased. The gene ontology (GO) analysis of biological process indicated that differentially expressed genes (DEGs) are genetically programmed to achieve immune and inflammatory pathways. Interestingly, we found the biphasic induction of various genes, including DAMPs and pro-inflammatory factors, peaking at acute and sub-acute phases. At the sub-acute phase, we also observed the induction of genes for phagocytosis as well as regulatory and growth factors. Further, we found the activation of CREB (cAMP-response element binding protein), one of the key players for neuronal plasticity, in peri-ischemic neurons by immunohistochemistry at this phase. Taken together, these findings raise the possibility the recurrent inflammation occurs at the sub-acute phase in the post-stroke brain, which could be involved in the debris clearance as well as neural reorganization.

White matter dissection and structural connectivity of the human vertical occipital fasciculus to link vision-associated brain cortex.

The vertical occipital fasciculus (VOF) is an association fiber tract coursing vertically at the posterolateral corner of the brain. It is re-evaluated as a major fiber tract to link the dorsal and ventral visual stream. Although previous tractography studies showed the VOF's cortical projections fall in the dorsal and ventral visual areas, the post-mortem dissection study for the validation remains limited. First, to validate the previous tractography data, we here performed the white matter dissection in post-mortem brains and demonstrated the VOF's fiber bundles coursing between the V3A/B areas and the posterior fusiform gyrus. Secondly, we analyzed the VOF's structural connectivity with diffusion tractography to link vision-associated cortical areas of the HCP MMP1.0 atlas, an updated map of the human cerebral cortex. Based on the criteria the VOF courses laterally to the inferior longitudinal fasciculus (ILF) and craniocaudally at the posterolateral corner of the brain, we reconstructed the VOF's fiber tracts and found the widespread projections to the visual cortex. These findings could suggest a crucial role of VOF in integrating visual information to link the broad visual cortex as well as in connecting the dual visual stream.

The external force associated with callus formation under the first metatarsal head is reduced by wearing rocker sole shoes.

INTRODUCTION: Callus is one of the main causes of diabetic foot ulcers. Therefore, preventing callus formation is very important. In a previous study, it was clarified that callus formation under the first metatarsal head (MTH) is associated with high shear stress time integral/pressure time integral (SPR-i). another study, it was clarified that rocker sole shoes are effective in reducing peak pressure under the first MTH. Therefore, we hypothesized that rocker sole shoes reduce SPR-i under the first MTH. This study aimed to clarify the effect of rocker sole shoes for external forces and leg motions in comparison with that of the normal sole shoes. METHODS: In-shoe external forces and leg motions were measured during walking wearing the normal sole shoes or the rocker sole shoes in healthy participants. As the external forces, the peak plantar pressure (PP), pressure time integral (PI), peak shear stress (PSS), and shear stress integral (SSI) of each gait cycle were calculated. Additionally, shear stress-pressure ratios (SPR) were calculated by dividing shear stress by pressure; concretely, peak values (SPR-p) and time integral values (SPR-i). As the leg motion, hip and knee joint motions were analyzed for the axis of flexion- extension. Three axes of ankle joint motion (inversion-eversion, plantar flexion-dorsiflexion, and adduction-abduction) were analyzed. RESULTS AND DISCUSSION: Six participants attended, and twelve feet were analyzed. When wearing the rocker sole shoes, the SPR-i under the first MTH was significantly smaller than when wearing the normal sole shoes. The SPR-i higher than 0.60 is associated with callus formation under the first MTH. In three of five feet with callus, SPR-i exceeded 0.60 in the normal sole shoes. The SPR-i of all three feet became smaller than 0.60 when wearing the rocker sole shoes. Although the knee (flexion-extension) and ankle (plantar flexion-dorsiflexion) joint motion became smaller when wearing the rocker sole shoes, there was no significant difference in walking speed. It is considered that propulsion was maintained by the push-off support provided by rocker sole shoes. CONCLUSION: It was suggested that rocker sole shoes are effective preventing callus formation under the first MTH.