Biology subjects

Tomita, S.

Publications and source records attributed to Tomita, S..

9 recordsLinked to original sources

bioRxiv2026

Mesoscopic cortical activities associated with pupil-linked perceptions inferred via explainable machine learning

Pupil dilation reflects arousal-related neural processes and is closely linked to sensory perception, attention, and cognitive state, but the mesoscopic cortical dynamics that accompany stimulus-evoked dilation remain unclear. Here, we combined simultaneous pupillometry and wide-field Ca{superscript 2} imaging in mice with explainable machine learning to identify cortical activity patterns predictive of pupil dilation. Cortical activity was recorded during hindpaw somatosensory stimulation, visual pattern change, and visual contextual stimulation using an visual oddball paradigm; spontaneous pupil dilation was additionally analyzed using a public resting-state dataset. To reduce multicollinearity in wide-field imaging signals, cortical activity was decomposed into independent components, which were used as inputs to recurrent neural network (RNN) decoders. Models reliably predicted sensory-evoked pupil dilation above shuffled-label controls when sessions with pre-existing pupil dilation were excluded, indicating that stimulus-evoked dilation was associated with distinct cortical activity patterns. Feature attribution using SHapley Additive exPlanations (SHAP) and spatial back-mapping revealed condition-dependent cortical signatures. Activity around the retrosplenial cortex and higher-order visual areas contributed to predictions during somatosensory and visual pattern stimulation, whereas secondary motor cortex activity was prominent during visual contextual stimulation. Spontaneous pupil dilation required lower-frequency cortical signals for decoding, suggesting partly distinct dynamics from sensory-evoked dilation. These findings demonstrate that explainable machine learning can extract candidate cortical activity patterns associated with pupil-linked arousal from high-dimensional wide-field Ca{superscript 2} imaging data. Because feature importance does not establish causality, the identified regions provide hypothesis-generating targets for future circuit manipulation studies.

neuroscience↗

bioRxiv2025

TARP γ-8 is a target of ethanol that regulates self-administration and relapse in mice

BackgroundBehavioral pathologies that characterize alcohol use disorder (AUD) are driven by the powerful reinforcing, or rewarding, properties of the drug. We have shown that glutamate AMPA receptor (AMPAR) activity is both necessary and sufficient for alcohol (ethanol) reinforcement. Transmembrane AMPAR regulatory protein (TARP) {gamma}-8 is an essential auxiliary protein that regulates AMPAR expression and activity; however, the role of TARP {gamma}-8 in AUD or other forms of addiction remains largely unexplored. ObjectivesThis study investigated the mechanistic role of TARP {gamma}-8 in operant ethanol self-administration (model of primary reinforcement) and cue-induced reinstatement of ethanol-seeking behavior (model of conditioned reinforcement) using TARP {gamma}-8 heterozygous null (+/-) mice. To determine if TARP {gamma}-8 signaling is targeted by ethanol use, we evaluated protein expression of TARP {gamma}-8, GluA1, CaMKII, and PSD-95 following ethanol self-administration. ResultsA battery of tests evaluating food and water intake, taste reactivity, anxiety-like behavior, and object recognition memory showed no fundamental behavioral deficits in TARP {gamma}-8 (+/-) mice, and no differences in response to acute ethanol or home-cage drinking as compared to wild-types. However, TARP {gamma}-8 (+/-) mice exhibited significantly reduced acquisition and escalation of operant ethanol self-administration and reduced cue-induced reinstatement of ethanol-seeking behavior, with no differences in parallel sucrose-only controls. In wild-type mice, ethanol self-administration increased TARP {gamma}-8 expression in the amygdala, nucleus accumbens, and hippocampus, and increased GluA1 expression in the amygdala and prefrontal cortex, compared to sucrose controls. ConclusionThese findings highlight the specificity of TARP {gamma}-8 regulation of ethanol reinforcement mechanisms and identify this crucial AMPAR auxiliary protein as a target of ethanol in reward-related brain regions, highlighting its potential for development of novel pharmacotherapies for AUD.

neuroscience↗

bioRxiv2024

Conserved loop of a phase modifier endows protein condensates with fluidity

Dipeptide repeats (DPRs) that are gene products from abnormal hexanucleotide repeat expansion in C9orf72 trigger amyotrophic lateral sclerosis (ALS) through unknown mechanism. This study highlights, importin Karyopherin{beta}2 (Kap{beta}2), which is responsible for nuclear transport and phase modification of RNA-binding proteins (RBPs), as a major DPR target. We demonstrate DPR accumulation in the nucleus via Kap{beta}2-mediated transport, which results in dose-dependent toxicity observed in nematode and yeast models. In vitro interaction studies exploiting chemical probe arrays and biophysical measurements reveal multivalent DPR binding to Kap{beta}2, including at the conserved acidic loop. Refractive index and fluorescence imaging coupled with biochemical assays unveiled that binding of excess DPRs to the acidic loop turns a phase modifier Kap{beta}2 into phase disrupter, resulting more condensed and viscous RBP condensates. Our findings provides molecular insight into C9orf72-ALS related to age and repeat expansion.

biochemistry↗

bioRxiv2024

Comparative metagenomic analysis provides novel insights into the ecophysiology of Myxococcota in activated sludge systems

Myxobacteria, belonging to the phylum Myxococcota, are ubiquitous in soil, marine, and other environments. Recent metagenomic sequencing analysis has shown that Myxococcota are predominant in activated sludge systems; however, their ecophysiological traits remain unclear. In this study, we evaluated the potential biological functions of 46 metagenomic bins of Myxococcota reconstructed from activated sludge samples from four municipal sewage treatment plants. The results showed that most Myxococcota bins had an almost complete set of genes associated with glycolysis and the TCA cycle. Palsa-1104 and Polyangiales bins contain glycoside hydrolase GH13 and peptidase M23, which are presumably involved in the lysis of the cell wall and cellular cytoplasm, indicating that part of the Myxococcota from activated sludge can prey on other microorganisms. The previously known cell contact-dependent predatory functions of Myxococcus xanthus are conserved in the family Myxococcaceae, though not in other families. Two bins belonging to Palsa-1104 had phototrophic gene clusters, suggesting that these bins may have both heterotrophic and autotrophic metabolisms. Assessment of the social behavior of the activated sludge Myxococcota, a FruA gene, and a C-signal gene involved in the regulation of fruiting body formation were lacking in Myxococcota bins, suggesting that they are incapable of fruiting body formation. In addition, multiple bins of Myxococcota had novel secondary metabolite biosynthesis gene clusters that may be used for the predation of other bacteria in the activated sludge. Our metagenome-based analyses provide novel insights into the microbial interactions associated with Myxococcota in activated sludge ecosystems.

microbiology↗

bioRxiv2023

A tryptophan metabolite modulates the host response to bacterial infection via kainate receptors

Bacterial infection involves a complex interaction between the pathogen and host where the outcome of infection is not solely determined by pathogen eradication. To identify small molecules that promote host survival by altering the host-pathogen dynamic, we conducted an in vivo chemical screen using zebrafish embryos and found that treatment with 3-hydroxy-kynurenine protects from lethal gram-negative bacterial infection. 3-hydroxy-kynurenine, a metabolite produced through host tryptophan metabolism, has no direct antibacterial activity but enhances host survival by restricting bacterial expansion in macrophages by targeting kainate-sensitive glutamate receptors. These findings reveal new mechanisms by which tryptophan metabolism and kainate-sensitive glutamate receptors function and interact to modulate immunity, with significant implications for the coordination between the immune and nervous systems in pathological conditions.

immunology↗

bioRxiv2023

A major endogenous glycosidase mediating quercetin uptake in Bombyx mori

Quercetin is a common plant flavonoid that is involved in herbivore-plant interactions. Mulberry silkworms (domestic silkworm, Bombyx mori, and wild silkworm, Bombyx mandarina) uptake quercetin from mulberry leaves and accumulate the metabolites in the cocoon, thereby improving its protective properties. Here we identified and characterized a glycosidase, named LPH-like quercetin glycoside hydrolase 1 (LQGH1), that initiates quercetin metabolism in the domestic silkworm. LQGH1 is expressed in the midgut where it mediates quercetin uptake by deglycosylating the three most common quercetin glycosides present in mulberry leaf: rutin, quercetin-3-O-malonyl-glucoside, quercetin-3-O-glucoside. Despite being located in an unequal crossing-over hotspot, LQGH1 is conserved in some species in clade Macroheterocera, including the wild silkworm, indicating the adaptive significance of quercetin uptake. LQGH1 is important also in breeding: defective mutations of LQGH1, which result in discoloration of the cocoon and increased silk yield, are homozygously conserved in 27 of the 32 Japanese white-cocoon domestic silkworm strains and 12 of the 30 Chinese ones we investigated.

genetics↗

bioRxiv2021

Partial endothelial-to-mesenchymal transition (EndMT) contributes to lumen re-organization after carotid arteryligation

Endothelial-to-mesenchymal transition (EndMT) is a fundamental process in vascular remodeling. Carotid artery ligation is commonly used for induction of neointima formation and vessel stenosis; however, the precise regulatory mechanism of vascular remodeling is not entirely understood. In this study, we showed that resident endothelial cells (ECs) are the origin of neointima cells and ECs transiently expressed CD45 in the early stage of neointima formation accompanied by increased expression of EndMT markers. In vitro, CD45-positive EndMT was induced by stabilization of HIF-1 with cobalt chloride or VHL inhibitor in human primary ECs, which mimicked the hypoxic condition of ligated artery, and promoted the formation of integrin 11-SHARPIN complex. Notably, a CD45 phosphatase inhibitor disrupted this complex, thereby destabilizing cell-cell junctions. These results suggest that the CD45 activity is required for the retention of an EC phenotype and cell-cell junctions during EndMT (termed "partial EndMT"). We thus propose a novel mechanism of partial EndMT that contributes to lumen re-organization during vascular injury.

molecular biology↗

bioRxiv2021

Activation of extrasynaptic kainate receptors drives hilar mossy cell activity

Mossy cells (MCs) of the dentate gyrus (DG) are key components of an excitatory associative circuit established by reciprocal connections with dentate granule cells (GCs). MCs are implicated in place field encoding, pattern separation and novelty detection, as well as in brain disorders such as temporal lobe epilepsy and depression. Despite their functional relevance, little is known about the determinants that control MC activity. Here, we examined whether MCs express functional kainate receptors (KARs), a subtype of glutamate receptors involved in neuronal development, synaptic transmission and epilepsy. Using mouse hippocampal slices, we found that bath application of submicromolar and micromolar concentrations of the KAR agonist kainic acid induced inward currents and robust MC firing. These effects were abolished in GluK2 KO mice, indicating the presence of functional GluK2-containing KARs in MCs. In contrast to CA3 pyramidal cells, which are structurally and functionally similar to MCs, and express synaptic KARs at mossy fiber (MF) inputs (i.e., GC axons), we found no evidence for KAR-mediated transmission at MF-MC synapses, indicating that most KARs at MCs are extrasynaptic. Immunofluorescence and immunoelectron microscopy analyses confirmed the extrasynaptic localization of GluK2-containing KARs in MCs. Finally, blocking glutamate transporters, a manipulation that increases extracellular levels of endogenous glutamate, was sufficient to induce KAR-mediated inward currents in MCs, suggesting that MC-KARs can be activated by increases in ambient glutamate. Our findings provide the first direct evidence of functional extrasynaptic KARs at a critical excitatory neuron of the hippocampus. Significance StatementHilar mossy cells (MCs) are an understudied population of hippocampal neurons that form an excitatory loop with dentate granule cells. MCs have been implicated in pattern separation, spatial navigation, and epilepsy. Despite their importance in hippocampal function and disease, little is known about how MC activity is recruited. Here, we show for the first time that MCs express extrasynaptic kainate receptors (KARs), a subtype of glutamate receptors critically involved in neuronal function and epilepsy. While we found no evidence for synaptic KARs in MCs, KAR activation induced strong action potential firing of MCs, raising the possibility that extracellular KARs regulate MC excitability in vivo and may also promote dentate gyrus hyperexcitability and epileptogenesis.

neuroscience↗

bioRxiv2017

Input-specific NMDAR-dependent potentiation of dendritic GABAergic inhibition

Preservation of a balance between synaptic excitation and inhibition is critical for normal brain function. A number of homeostatic cellular mechanisms have been suggested to play a role in maintaining this balance, including long-term plasticity of GABAergic inhibitory synapses. Many previous studies have demonstrated a coupling of postsynaptic spiking with modification of perisomatic inhibition. Here, we demonstrate that activation of NMDA-type glutamate receptors leads to input-specific long-term potentiation of dendritic inhibition mediated by somatostatin-expressing interneurons. This form of plasticity is expressed postsynaptically and requires both CaMKII and the {beta}2-subunit of the GABA-A receptor. Importantly, this process may function to preserve dendritic inhibition, as in vivo loss of NMDAR signaling results in a selective weakening of dendritic inhibition. Overall, our results reveal a new mechanism for linking excitatory and inhibitory input in neuronal dendrites and provide novel insight into the homeostatic regulation of synaptic transmission in cortical circuits.

neuroscience↗