Biology subjects

Koizumi, A.

Publications and source records attributed to Koizumi, A..

12 recordsLinked to original sources

bioRxiv2026

Novel cell wall-associated genes that enable Cryptococcus neoformans to evade dectin-1-mediated innate immune recognition.

The fungal pathogen Cryptococcus neoformans contains about 200 {micro}g of {beta}-1,3-glucan (1,3BG) per 1 mg of dry cell weight when grown under standard culture conditions (YPD medium at 30 {degrees}C under aerobic conditions). However, 1,3BG exposure is tightly suppressed, even in capsule-deficient strains, allowing the fungus to evade recognition by the immune receptor dectin-1 and anti-1,3BG antibodies. Although other pathogenic fungi mask 1,3BG with -1,3-glucan (1,3AG) to evade dectin-1 recognition, the factors responsible for 1,3BG masking and dectin-1 evasion in C. neoformans remain incompletely understood. To identify capsule-independent 1,3BG masking and dectin-1-evasion factors, we generated a series of cell wall-related gene deletion strains in the capsule-deficient strain cap59{Delta} by using CRISPR-Cas9 and screened for mutants that failed to evade dectin-1 binding. We found eight deletants (cap59{Delta}/mpk1{Delta}, cap59{Delta}/chs3{Delta}, cap59{Delta}/kre5{Delta}, cap59{Delta}/crz1{Delta}, cap59{Delta}/kre6{Delta}/skn1{Delta}, cap59{Delta}/hxl1{Delta}, cap59{Delta}/uge1{Delta}, and cap59{Delta}/ugt1{Delta}) that exhibited increased binding to dectin-1 or anti-1,3BG antibody, or both. As a similar phenotype was not observed in cap59{Delta}/ags1{Delta}, 1,3AG-mediated masking of 1,3BG appears to play a limited role in C. neoformans. These eight deletants induced significantly greater secretion of IL-6 and IL-1{beta} from dendritic cells (DCs) than did cap59{Delta}. This enhanced inflammatory response was markedly attenuated in dectin-1-deficient DCs, indicating that the increased immunogenicity was driven by 1,3BG exposure and subsequent dectin-1 recognition. Collectively, these findings demonstrate that multiple genes involved in maintaining cell wall integrity--including those involved in {beta}-1,6-glucan and chitosan biosynthesis--are essential for regulating 1,3BG exposure and enabling C. neoformans to evade dectin-1-mediated immune recognition. HighlightsO_LIWe identified new capsule-independent {beta}-1,3-glucan-masking genes in C. neoformans C_LIO_LIGene deletants had higher dectin-1 deposition than parental acapsular cap59{Delta} C_LIO_LIDeletion of these genes enhanced IL-6 and IL-1{beta} secretion by dendritic cells C_LIO_LIThe enhanced cytokine response was suppressed in dendritic cells lacking dectin-1. C_LIO_LIDeletant strains may serve as new whole-cell antigens for cryptococcal vaccines C_LI

microbiology↗

bioRxiv2026

HAC1 contributes to stress adaptation and virulence in the emerging fungal pathogen Candida auris

Candida auris (Candidozyma auris) is an emerging multidrug-resistant fungal pathogen that poses a significant global health threat. However, the molecular mechanisms underlying its virulence remain incompletely understood. In this study, we performed in vivo transcriptome analysis using an immunosuppressed mouse gastrointestinal infection model to identify genes associated with host-adaptation and virulence during infection. By comparing fungal transcriptomes obtained from colonization and dissemination sites with those from in vitro cultures, we identified genes that were consistently upregulated during infection. Among these genes, the unfolded protein response regulator HAC1 was selected as a candidate virulence-associated gene for further analysis. RT-PCR and sequencing analyses revealed that HAC1 mRNA in C. auris undergoes an unconventional splicing event of 287 bp that is enhanced under ER stress conditions. The excised region spans the annotated open reading frame boundary, suggesting that the translated region of HAC1 may require re-evaluation. Notably, a proportion of HAC1 transcripts appeared to be spliced even under non-stress conditions, indicating a detectable basal level of UPR activation. Differences in splicing dynamics were also observed among clade strains. Functional analyses demonstrated that deletion of HAC1 increased sensitivity to ER stress and heat stress. The HAC1 deletion mutant also exhibited reduced virulence in both Galleria mellonella and immunosuppressed mouse infection models, as evidenced by delayed host mortality and decreased fungal burdens, respectively. These findings indicate that HAC1 contributes to ER stress adaptation, thermotolerance, and survival in the host environment, and identify HAC1 as a virulence-associated gene in C. auris.

microbiology↗

bioRxiv2023

Time-dependent neural arbitration between cue associative and episodic fear memories

After traumatic events, simple cue-threat associative memories strengthen while episodic memories become fragmented. However, how the brain prioritizes cue associations over episodic coding of traumatic events remains unclear. Here, we developed a new episodic threat conditioning paradigm in which participants concurrently form two memory representations: cue associations and episodic cue sequence. We discovered that these two distinct memories compete for physiological fear expression, reorganizing overnight from an overgeneralized cue-based to a precise sequence-based expression. With multivariate fMRI, we track inter-area communication of the memory representations and demonstrate that a shift from hippocampal-dominant to prefrontal-dominant control of the fear regulatory circuit governs this memory maturation. Critically, this overnight reorganization is altered in individuals with heightened trait anxiety. Together, these findings suggest the brain prioritizes generalizable associative memories under recent traumatic stress, but resorts to selective episodic memories 24 hrs later. Time-dependent memory competition provides a unifying account for memory dysfunctions in posttraumatic stress disorders.

neuroscience↗

bioRxiv2022

Cleavage of α-1,4-Glycosidic Linkages by the Glycosylphosphatidylinositol-Anchored α-Amylase AgtA Decreases the Molecular Weight of Cell Wall α-1,3-Glucan in Aspergillus oryzae

Aspergillus fungi contain -1,3-glucan with a low proportion of -1,4-glucan as a major cell wall polysaccharide. Glycosylphosphatidylinositol (GPI)-anchored -amylases are conserved in Aspergillus fungi. The GPI-anchored -amylase AmyD in Aspergillus nidulans has been reported to directly suppress the biosynthesis of cell wall -1,3-glucan but not to degrade it in vivo. However, the detailed mechanism of cell wall -1,3-glucan biosynthesis regulation by AmyD remains unclear. Here we focused on AoAgtA, which is encoded by the Aspergillus oryzae agtA gene, an ortholog of the A. nidulans amyD gene. Similar to findings in A. nidulans, agtA overexpression in A. oryzae grown in submerged culture decreased the amount of cell wall -1,3-glucan and led to the formation of smaller hyphal pellets in comparison with the wild-type strain. We analyzed the enzymatic properties of recombinant (r)AoAgtA produced in Pichia pastoris and found that it degraded soluble starch, but not linear bacterial -1,3-glucan. Furthermore, rAoAgtA cleaved 3--maltotetraosylglucose with a structure similar to the predicted boundary structure between the -1,3-glucan main chain and a short spacer composed of -1,4-linked glucose residues in cell wall -1,3-glucan. Interestingly, rAoAgtA randomly cleaved only the -1,4-glycosidic bonds of 3--maltotetraosylglucose, indicating that AoAgtA may cleave the spacer in cell wall -1,3-glucan. Consistent with this hypothesis, heterologous overexpression of agtA in A. nidulans decreased the molecular weight (MW) of cell wall -1,3-glucan. These in vitro and in vivo properties of AoAgtA suggest that GPI-anchored -amylases can degrade the spacer -1,4-glycosidic linkages in cell wall -1,3-glucan before its insolubilization, and this spacer cleavage decreases the MW of cell wall -1,3-glucan in vivo.

microbiology↗

bioRxiv2022

Fear in action: Fear conditioning and alleviation through body movements

Acquisition of fear memories enhances survival especially when the memories guide defensive movements to minimize harm. Accordingly, fear memories and body movements have tight relationships in animals: Fear memory acquisition results in adapting reactive defense movements, while training active defense movements to avoid threat reduces fear memory. However, evidence in humans is scarce because their movements are typically marginalized in experiments. Here, we tracked participants whole-body motions while they underwent fear conditioning in a virtual 3D space. First, representational similarity analysis of body motions revealed that participants obtained distinct spatiotemporal movement patterns through fear conditioning. Second, subsequent training to actively avoid threats with naturalistic defensive actions led to a long-term (24 hrs) reduction of physiological and embodied conditioned responses, while extinction or vicarious training only transiently reduced the responses followed by their spontaneous return. Together, our results highlight the intrinsic role of body movements in human fear memory functions, suggesting the potential for improving fear memory interventions through embodiment.

neuroscience↗

bioRxiv2021

A glycosylphosphatidylinositol-anchored α-amylase encoded by amyD contributes to a decrease in the molecular mass of cell wall α-1,3-glucan in Aspergillus nidulans

-1,3-Glucan is one of the main polysaccharides in the cell wall of Aspergillus nidulans. We previously revealed that it plays a role in hyphal aggregation in liquid culture, and that its molecular mass (MM) in an agsA-overexpressing (agsAOE) strain was larger than that in an agsB-overexpressing (agsBOE) strain. The mechanism that regulates the MM of -1,3-glucan is poorly understood. Although the gene amyD, which encodes glycosyl-phosphatidylinositol (GPI)-anchored -amylase (AmyD), is involved in the biosynthesis of -1,3-glucan in A. nidulans, how it regulates this biosynthesis remains unclear. Here we constructed strains with disrupted amyD ({Delta}amyD) or overexpressed amyD (amyDOE) in the genetic background of the ABPU1 (wild-type), agsAOE, or agsBOE strain, and characterized the chemical structure of -1,3-glucans in the cell wall of each strain, focusing on their MM. The MM of -1,3-glucan from the agsBOE amyDOE strain was smaller than that in the parental agsBOE strain. In addition, the MM of -1,3-glucan from the agsAOE {Delta}amyD strain was greater than that in the agsAOE strain. These results suggest that AmyD is involved in decreasing the MM of -1,3-glucan. We also found that the C-terminal GPI-anchoring region is important for these functions.

microbiology↗

bioRxiv2020

Multivoxel Neural Reinforcement Changes Resting-State Functional Connectivity Within the Threat Regulation Network

Using neural reinforcement, participants can be trained to pair a reward with the activation of specific multivoxel patterns in their brains. In a double-blind placebo-controlled experiment, we previously showed that this intervention can decrease the physiological reactivity associated with naturally feared animals. However, the mechanisms behind the effect remain incompletely understood and its usefulness for treatment remains unclear. If the intervention fundamentally changed the brain responses, we might expect to observe relatively stable changes in the functional connectivity within the threat regulation network. To evaluate this possibility, we conducted functional magnetic resonance imaging (fMRI) sessions while subjects were at rest, before and after neural reinforcement, and quantified the changes in resting-state functional connectivity accordingly. Our results indicate that neural reinforcement increased the connectivity of prefrontal regulatory regions with the amygdala and the ventral temporal cortex (where the visual representations of phobic targets are). Surprisingly, we found no evidence of Hebbian-like learning during neural reinforcement, contrary to what one may expect based on previous neurofeedback studies. These results suggest that multivoxel neural reinforcement, also known as decoded neurofeedback (DecNef), may operate via unique mechanisms, distinct from those involved in conventional neurofeedback.

neuroscience↗

bioRxiv2019

Atypical spatial frequency dependence of visual metacognition among schizophrenia patients

Although altered early stages of visual processing have been reported among schizophrenia patients, how such atypical visual processing may affect higher-level cognition remains largely unknown. Here we tested the hypothesis that metacognitive performance may be atypically modulated by spatial frequency (SF) of visual stimuli among individuals with schizophrenia, given their altered magnocellular function. To study the effect of SF on metacognitive performance, we asked patients and controls to perform a visual detection task on gratings with different SFs and report confidence, and analyzed the data using the signal detection theoretic measure meta-d. Control subjects showed better metacognitive performance after yes- (stimulus-presence) than after no- (stimulus-absence) responses ( yes-response advantage) for high SF (HSF) stimuli but not for low SF (LSF) stimuli. The patients, to the contrary, showed a yes-response advantage not only for HSF but also for LSF stimuli, indicating atypical SF dependency of metacognition. An fMRI experiment using the same task revealed that the dorsolateral prefrontal cortex (DLPFC), known to be crucial for metacognition, shows activity mirroring the behavioral results: decoding accuracy of perceptual confidence in DLPFC was significantly higher for HSF than for LSF stimuli in controls, whereas this decoding accuracy was independent of SF in patients. While individuals without schizophrenia may flexibly adapt metacognitive computations across SF ranges, patients may employ a different mechanism that is independent of SF. Because visual stimuli of low SF have been linked to top-down processing in predictive coding, this may reflect atypical functioning in these processes in schizophrenia. HighlightsO_LIVisual metacognition of controls was dependent on spatial frequency. C_LIO_LIVisual metacognition of schizophrenia patients was independent of spatial frequency. C_LIO_LIPatients and controls differently rely on the dorsolateral prefrontal cortex. C_LIO_LISensory inputs may reach metacognitive circuits in an atypical manner among patients. C_LI

neuroscience↗

bioRxiv2019

A reciprocal inhibition model of alternations between non-dissociative and dissociative states in patients with PTSD

ObjectiveTraumatic life-events can leave individuals with contrasting posttraumatic stress disorder (PTSD) symptoms, including re-experiencing and avoidance. Notably, patients with PTSD are known to periodically switch between two opposing attentional biases; namely, toward threat and away from threat. We hypothesized that reciprocal inhibition between the amygdala and ventromedial prefrontal cortex (vmPFC) may induce alternations between these attentional biases, which in turn may contribute to the re-experiencing and avoidance symptoms, respectively.\n\nMethodsTo test this reciprocal inhibition model, we performed an experiment to measure the attentional biases of patients with PTSD. We examined the differential relationships between PTSD symptom clusters (re-experiencing/avoidance) and attentional biases (toward/away from threat). Additionally, we performed a meta-regression analysis to examine the role of amygdala reactivity in the imbalance between re-experiencing and avoidance symptoms.\n\nResultsWe found that attentional bias toward threat was selectively associated with re-experiencing symptoms whereas attentional bias away from threat was selectively associated with avoidance symptoms. Meta-regression analysis based on twelve participant populations (total N = 316) revealed that left amygdala activity was positively correlated with the severity of re-experiencing symptoms relative to avoidance symptoms.\n\nConclusionsOur findings support the hypothesis that reciprocal inhibition of common neural circuits may underlie the switch between attentional biases toward and away from threat as well as that between re-experiencing and avoidance symptoms. Re-experiencing and avoidance/emotional numbing are the core symptoms used to distinguish between the non-dissociative and dissociative PTSD subtypes. The reciprocal inhibition mechanism may help elucidate the mechanisms underlying those PTSD subtypes.

neuroscience↗

bioRxiv2019

An asexual flower of Silene latifolia and Microbotryum lychnidis-dioicae promoting its sexual-organ development

Silene latifolia is a dioecious flowering plant with sex chromosomes in the family Caryophyllaceae. Development of a gynoecium and stamens are suppressed in the male and female flowers of S. latifolia, respectively. Microbtryum lychnidis-dioicae promotes stamen development when it infects the female flower. If suppression of the stamen and gynoecium development is regulated by the same mechanism, suppression of gynoecium and stamen development is released simultaneously with the infection by M. lychnidis-dioicae. To assess this hypothesis, an asexual mutant, without gynoecium or stamen, was infected with M. lychnidis-dioicae. A filament of the stamen in the infected asexual mutant was elongated at stages 11 and 12 of the flower bud development as well as the male, but the gynoecium did not form. Instead of the gynoecium, a filamentous structure was suppressed as in the male flower. Developmental suppression of the stamen was released by M. lychnidis-dioicae, but that of gynoecium development was not released. It is thought, therefore, that the suppression of gynoecium development was not released by the infection of M. lychnidis-dioicae. M. lychnidis-dioicae would have a function similar to SPF since the elongation of the stamen that is not observed in the healthy asexual mutant was observed after stage 8 of flower bud development. Such an infection experiment also that the Y chromosome of the asexual mutant has genes related to the differentiation of archesporial cells, but none related to maturation of the tapetal cells.

plant biology↗

bioRxiv2019

Both galactosaminogalactan and α-1,3-glucan contribute to aggregation of Aspergillusoryzae hyphae in liquid culture

Filamentous fungi generally form aggregated hyphal pellets in liquid culture. We previously reported that -1,3-glucan-deficient mutants of Aspergillus nidulans did not form hyphal pellets and their hyphae were fully dispersed, and we suggested that -1,3-glucan functions in hyphal aggregation. Yet, Aspergillus oryzae -1,3-glucan-deficient (AG{Delta}) mutants still form small pellets; therefore, we hypothesized that another factor responsible for forming hyphal pellets remains in these mutants. Here, we identified an extracellular matrix polysaccharide galactosaminogalactan (GAG) as such a factor. To produce a double mutant of A. oryzae (AG-GAG{Delta}), we disrupted the genes required for GAG biosynthesis in an AG{Delta} mutant. Hyphae of the double mutant were fully dispersed in liquid culture, suggesting that GAG is involved in hyphal aggregation in A. oryzae. Addition of partially purified GAG fraction to the hyphae of the AG-GAG{Delta} strain resulted in formation of mycelial pellets. Acetylation of the amino group in galactosamine of GAG weakened GAG aggregation, suggesting that hydrogen bond formation by this group is important for aggregation. Genome sequences suggest that -1,3-glucan, GAG, or both are present in many filamentous fungi and thus may function in hyphal aggregation in these fungi. We also demonstrated that production of a recombinant polyesterase, CutL1, was higher in the AG-GAG{Delta} strain than in the wild-type and AG{Delta} strains. Thus, controlling hyphal aggregation factors of filamentous fungi may increase productivity in the fermentation industry.\n\nImportanceProduction using filamentous fungi is an important part of the fermentation industry, but hyphal aggregation in these fungi in liquid culture limits productivity compared with that of yeast or bacterial cells. We found that galactosaminogalactan and -1,3-glucan both function in hyphal aggregation in Aspergillus oryzae, and that the hyphae of a double mutant deficient in both polysaccharides become fully dispersed in liquid culture. We also revealed the relative contribution of -1,3-glucan and galactosaminogalactan to hyphal aggregation. Recombinant protein production was higher in the double mutant than in the wild-type strain. Our research provides a potential technical innovation for the fermentation industry that uses filamentous fungi, as regulation of the growth characteristics of A. oryzae in liquid culture may increase productivity.

cell biology↗

bioRxiv2018

Layer-specific ultra-high field 7T fMRI showing that threat anticipation is mediated by the pulvinar input to the superficial layers of primary visual cortex (V1)

The perceptual system gives priority to threat-relevant signals with survival value. Its mechanism may not only include the processing initiated in the presence of threat signals but also in the mere anticipation of such signals. Here, we show that the pulvinar modulates activity in the early visual cortex (V1) specifically in threat anticipation. Using ultra-high-field 7T fMRI, we examined the layer-specific interaction between V1 and the pulvinar, while taking advantage of the fact that the directionality of such interaction is anatomically constrained in specific V1 layers. Only in anticipation of a fearful face target, but not of a control happy face target, was false perception of anticipated-yet-unpresented target face accompanied by stronger activity in the V1 superficial-cortical-depth (layers 1-3), which was preceded by pre-target-onset pulvinar activity. The pulvinar may contribute to the visual processing initiated in the anticipation of threat, and play an important role in anxiety.

neuroscience↗