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Yasuda, H.

Publications and source records attributed to Yasuda, H..

5 recordsLinked to original sources

Repeated swim exposure and PKN1a knockout enhance group I mGluR-dependent excitability associated with reduced EAAT3 expression in mouse dentate granule cells

Stress-related experiences alter glutamatergic signaling and neuronal excitability, but the mechanisms that couple experience to dentate granule cell function remain incompletely understood. Here, we examined how protein kinase N1a (PKN1a), a protein kinase C-like serine/threonine kinase, and repeated swim exposure regulate mouse hippocampal dentate granule cell excitability, with a focus on the neuronal glutamate transporter excitatory amino acid transporter 3 (EAAT3) and group I metabotropic glutamate receptors (mGluRs). Five days of repeated swim exposure increased spike firing in mature dentate granule cells from wild-type mice. PKN1a knockout produced a similar increase, and repeated swim did not further enhance firing in knockout mice. The enhanced firing observed after repeated swim exposure and in PKN1a knockout mice was reduced by co-application of an mGluR1 antagonist (LY367385) and an mGluR5 antagonist (MPEP). Inhibition of glutamate transporters with DL-TBOA increased granule cell firing in control wild-type mice but did not further increase firing in repeated-swim wild-type or PKN1a knockout mice, suggesting occlusion of transporter-dependent regulation of excitability. Repeated swim exposure and PKN1a knockout also reduced total and surface expression of EAAT3 in the hippocampus, whereas expression of the glial glutamate transporter EAAT2 was not significantly altered. Finally, PKN1a knockout and repeated swim exposure reduced anxiety-related behavior in the elevated plus maze test. Thus, PKN1a-dependent regulation of EAAT3 may restrain group I mGluR-dependent excitability in dentate granule cells, whereas repeated swim exposure and PKN1a knockout shift this system toward a lower-EAAT3, higher-excitability state accompanied by reduced anxiety-related behavior.

neuroscience↗

Differential physiological, behavioral, and medial prefrontal cortex transcriptomic responses to chronic restraint stress between BALB/c and C57BL/6J mice

Chronic stress is a major risk factor for psychiatric disorders such as depression and anxiety, yet the biological basis of individual differences in stress susceptibility and resilience remains poorly understood. Here, we examined physiological, behavioral, and medial prefrontal cortex (mPFC) transcriptomic responses to chronic restraint stress (CRS) in male BALB/c and C57BL/6J mice. After 21 days of CRS, BALB/c mice exhibited greater stress-related changes than C57BL/6J mice, including greater body weight loss, elevated serum corticosterone, reduced serum antioxidant capacity, and more pronounced depression-like behaviors. RNA sequencing showed largely strain-specific transcriptional changes in the mPFC. Strain x stress interaction analysis, followed by canonical pathway analysis using Ingenuity Pathway Analysis (IPA), identified strain-dependent molecular signatures. The most prominent differences involved extracellular matrix (ECM) organization and remodeling and neuroinflammatory signaling pathways, with greater predicted activation in BALB/c mice. IPA upstream regulator analysis further predicted multiple candidate regulators associated with these pathways, including TGF-{beta}/SMAD, C4a/C4b, and MAPK14. Among genes associated with these pathways, several ECM-related genes were preferentially upregulated in BALB/c mice, whereas activity-dependent immediate early genes were preferentially downregulated in C57BL/6J mice. These findings suggest that the strain-dependent mPFC transcriptional programs identified here may contribute to differential stress susceptibility and resilience.

neuroscience↗

PCP-dependent polarity propagation across neuronal columns in the Drosophila medulla

Planar cell polarity (PCP) signaling establishes coordinated tissue polarity through asymmetric localization of core PCP factors and propagation of polarity information between neighboring cells. While PCP mechanisms are well characterized in continuous epithelial tissues, whether PCP-dependent polarity propagation can operate across discontinuous neuronal structures remains unclear. During pupal development of the Drosophila medulla, R8 photoreceptor axon terminals transiently form horseshoe-shaped structures whose orientation reflects neuronal column polarity. Here, we show that Frizzled (Fz) progressively becomes asymmetrically localized within developing horseshoe structures, whereas Van Gogh (Vang) transiently accumulates in adjacent glial cells during early developmental stages. Conditional disruption of Vang in glial cells impaired asymmetric Fz localization and disrupted horseshoe orientation polarity. Glial Vang localization preceded the emergence of robust Fz asymmetry, suggesting that glial Vang functions during an early polarity establishment phase. Furthermore, mosaic knockdown of fz in R8 neurons altered Fz localization in neighboring R8 terminals, suggesting local propagation of polarity information between adjacent neuronal columns. Together, our results suggest that PCP-dependent polarity propagation operates across discontinuous neuronal columns in the developing medulla and provide a framework for understanding coordinated polarity formation in neural tissues lacking continuous epithelial organization.

developmental biology↗

14-3-3 proteins stabilize actin and vimentin filaments to maintain processes in glomerular podocyte

Adaptor protein 14-3-3s have isoform-specific binding partners and roles. We reported 14-3-3{beta} interacts with FKBP12 and synaptopodin to maintain the structure of actin fibers in podocytes. However, differential roles of 14-3-3 isoforms in kidneys are unclear. Herein, we showed that 14-3-3{beta} was dominantly co-localized with FKBP12 in foot processes and was partially co-localized with Par3 at slit diaphragm in podocytes. 14-3-3{beta} interacted with Par3, and FKBP12 bound to 14-3-3{beta} competitively with Par3. Although deletion of 14-3-3{beta} enhanced the interaction of Par3-Par6, it altered actin fiber structure and processes. 14-3-3{beta} and synaptopodin were downregulated in podocyte injury models. 14-3-3{sigma} in podocytes interacted with vimentin in primary processes but not with the actin-associated proteins in foot processes. Deletion of 14-3-3{sigma} altered vimentin fiber structure and processes. 14-3-3{sigma} and vimentin were upregulated in the early phase of podocyte injury models but were decreased in the end stage. Together, the precise localization of 14-3-3{beta} at actin cytoskeleton plays a role in maintaining foot processes and Par complex in podocytes. 14-3-3{sigma} at vimentin cytoskeleton is essential for maintaining primary processes.

cell biology↗

Nasal delivery of single-domain antibodies improve symptoms of SARS-CoV-2 infection in an animal model

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that causes the disease COVID-19 can lead to serious symptoms, such as severe pneumonia, in the elderly and those with underlying medical conditions. While vaccines are now available, they do not work for everyone and therapeutic drugs are still needed particularly for treating life-threatening conditions. Here, we showed nasal delivery of a new, unmodified camelid single-domain antibody (VHH), termed K-874A, effectively inhibited SARS-CoV-2 titers in infected lungs of Syrian hamsters without causing weight loss and cytokine induction. In vitro studies demonstrated that K-874A neutralized SARS-CoV-2 in both VeroE6/TMPRSS2 and human lung-derived alveolar organoid cells. Unlike other drug candidates, K-874A blocks viral membrane fusion rather than viral attachment. Cryo-electron microscopy revealed K-874A bound between the receptor binding domain and N-terminal domain of the virus S protein. Further, infected cells treated with K-874A produced fewer virus progeny that were less infective. We propose that direct administration of K-874A to the lung via a nebulizer could be a new treatment for preventing the reinfection of amplified virus in COVID-19 patients. Author summaryVaccines for COVID-19 are now available but therapeutic drugs are still needed to treat life-threatening cases and those who cannot be vaccinated. We discovered a new heavy-chain single-domain antibody that can effectively neutralize the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that causes COVID-19. Unlike other drug candidates, which prevent the virus from attaching to the receptor on the host cell, this new antibody acts by blocking the virus membrane from fusing with the host cell membrane. We studied the behavior of the new antibody in vitro using VeroE6/TMPRSS2 cells and human lung organoids. When delivered through the nose to infected Syrian hamsters, we found that this antibody could prevent the typical symptoms caused by SARS-CoV-2. Our results are significant because delivering simple drugs directly to infected lungs using a nebulizer could increase the potency of the drugs while reducing the risk of immune reaction that could occur if the drugs escape or are delivered through the blood stream.

microbiology↗