Biology subjects

Takeuchi, K.

Publications and source records attributed to Takeuchi, K..

At least 19 recordsLinked to original sources

bioRxiv2026

CLOSING THE CONFORMATIONAL CYCLE OF THE BETA-BARREL ASSEMBLY MACHINERY

The {beta}-barrel assembly machinery (BAM) catalyzes the folding and insertion of {beta}-barrel outer membrane proteins (OMPs) in Gram-negative bacteria without an external energy source. BAM comprises the central subunit BamA and associated lipoproteins, with BamA forming a conformationally dynamic lateral gate. Although substrate-engaged structures have revealed key steps in OMP assembly, how substrate-free BamA closes its lateral gate and which conformations it adopts in a lipid bilayer remain unclear. Here, we determined cryo-EM structures of the Escherichia coli (E. coli) BAM complex in three detergents and in Saposin A nanodiscs composed of E. coli polar lipids. In addition to the canonical outward-open (OO) state, we identified an occluded (OC) state in which both the lateral gate and the periplasmic opening of the BamA barrel are closed. OC predominated in lipid-reconstituted BAM that retained OMP-folding activity, indicating that this state is substantially populated in a functional BAM ensemble within a lipid bilayer. We also identified a semi-occluded (sOC) state in which {beta}1-{beta}2 adopts an OC-like closed position while rearrangement of {beta}16 remains incomplete. Comparison of OO, sOC, and OC reveals asynchronous rearrangements of the {beta}1-{beta}2 and {beta}16 regions, with {beta}1-{beta}2 movement occurring without complete repositioning of {beta}16. Re-examination of previously reported lipid-embedded BAM structures further highlights the importance of local map quality at the gate region for conformational assignment. Together, our findings expand the BamA conformational landscape beyond the conventional OO-inward-open (IO) framework to include OC and sOC as distinct structural states associated with lateral-gate closure following OMP insertion.

molecular biology↗

bioRxiv2026

Tissue-resolved photosynthetic responses in living leaves revealed by microscopic imaging-pulse-amplitude-modulation

Leaves of terrestrial plants possess heterogeneous anatomical structures composed of multiple cell layers. Although biochemical differences among leaf tissues have been inferred from protein analyses and anatomical studies, direct comparisons of dynamic photochemical responses among tissues while preserving their spatial context remain challenging. Leaves experience intrinsically heterogeneous environments because incident light enters primarily from above and propagates through complex internal leaf structures. Therefore, analyzing photosynthetic activity at the tissue and cellular levels is essential for understanding how photosynthesis operates within structurally heterogeneous leaves. Here, we combined live leaf-section imaging with microscopic Imaging-PAM chlorophyll fluorescence measurements to analyze photochemical responses at the tissue level in living leaf sections. In dorsiventral dicot leaves, palisade tissues exhibited a higher effective PSII quantum yield [Y(II)] and more rapid induction of regulated energy dissipation [Y(NPQ)] than spongy tissues, indicating higher photosynthetic capacity and photoprotective activity. In contrast, rice leaves, which lack palisade-spongy differentiation, showed uniform photochemical responses along the adaxial-abaxial axis. In the C4 plant finger millet, mesophyll and bundle sheath cells displayed distinct photochemical responses consistent with their functional differentiation in C4 photosynthesis. These results demonstrate that photosynthetic responses are spatially organized according to leaf anatomical structure. Although section-based measurements do not reproduce the native optical and gas environments of intact leaves, they enable the comparison of intrinsic tissue-specific photochemical properties under approximately equivalent illumination. The present approach enables tissue-resolved chlorophyll fluorescence analysis in living leaves, providing a new framework for investigating how leaf architecture shapes the spatial organization of photosynthetic activity.

plant biology↗

bioRxiv2026

The PSI-NDH supercomplex prevents chilling-induced PSI photoinhibition

Chilling stress induces photosystem I (PSI) photoinhibition in chilling-sensitive cucumber, in which insufficient activity of the chloroplast NADH dehydrogenase-like complex (NDH) leads to PSI over-reduction and damage. However, it is not yet clear whether these findings can be generalized to other species or what the molecular mechanism underlying impaired NDH function is. In this study, we first examined whether NDH is essential for PSI protection under chilling stress using an NDH-deficient rice mutant. Compared with wild-type plants, the NDH-deficient mutant exhibited enhanced PSI over-reduction and pronounced PSI photoinhibition under chilling stress. In contrast, rice plants expressing flavodiiron protein (FLV), which functions as an alternative electron acceptor downstream of PSI, did not exhibit PSI photoinhibition under chilling stress, demonstrating that electron sink capacity of NDH is important for PSI protection under chilling stress. Furthermore, analysis of the factors responsible for NDH dysfunction under chilling stress in cucumber revealed that chilling stress destabilizes the PSI-NDH supercomplex, leading to NDH monomerization and a consequent loss of NDH activity. This NDH monomerization is likely attributable to chilling-induced damage to the light-harvesting complex Lhca, which mediates the association between PSI and NDH. Together, these results indicate that NDH is essential for protecting PSI from photoinhibition under chilling stress in both rice and cucumber, and that chilling-induced destabilization of the PSI-NDH supercomplex represents a key molecular mechanism underlying PSI over-reduction and photoinhibition.

plant biology↗

bioRxiv2026

Tumor-associated tissue-resident macrophages drive pancreatic cancer progression through IGF1-IGF1R signaling

The specific contribution of tissue-resident macrophages (TRMs) to pancreatic ductal adenocarcinoma (PDAC) progression remains unclear. Here, we found that a high abundance of TRM-derived tumor-associated macrophages (TRM-TAMs) is an independent indicator of poor prognosis in patients with PDAC. To elucidate the underlying mechanism, we established an advanced organoid platform (iMac-FPCO), which incorporates macrophages derived from human induced pluripotent stem cells to reflect the differentiation process of TRMs. Single-cell transcriptomic analysis revealed this model recapitulates the transcriptional identity of TRM-TAMs in patient tissue. We demonstrated that TRM-TAMs drive cancer cell proliferation, while maintaining chemoresistance, and identified TRM-derived insulin-like growth factor 1 (IGF1) as the critical mediator. This result provides a rationale for why previous trials targeting IGF1 receptor (IGF1R) failed to improve survival in unselected patient populations. We hypothesize that stratifying patients by TRM-TAM abundance could help identify a responsive subgroup, thereby reviving IGF1R-targeted therapy as a viable treatment for PDAC.

cancer biology↗

bioRxiv2026

Spatial proteomics reveals prefrontal circuit diversity in socioemotional behaviour

Understanding how molecular diversity across long-range neural circuits governs brain function remains a central challenge in neuroscience. Here, we developed a projection-specific spatial proteomics approach and revealed robust presynaptic molecular divergence across six projection-defined pathways of the medial prefrontal cortex (mPFC), including efferent projections to the basolateral amygdala (BLA), nucleus accumbens (NAc), thalamus (Thal), hypothalamus (HT) and cortex (CTX), as well as the afferent projection from the BLA to the mPFC. Among these pathways, we identify BLTP2 (KIAA0100), a previously uncharacterized transmembrane protein, as highly enriched in the projection from the mPFC to the BLA. BLTP2 localizes to excitatory presynaptic terminals and is enriched at synapses that are activated during memory formation. Loss of BLTP2 impairs synaptic structure and transmission, and reduces activity-dependent remodelling, resulting in selective deficits in contextual fear memory, anxiety-related behavior, and social behavior. Mechanistically, BLTP2 promotes presynaptic assembly by recruiting Neurexin 1. These findings reveal projection-specific presynaptic molecular diversity and provide mechanistic insights into circuit-level vulnerabilities in neuropsychiatric disorders.

neuroscience↗

bioRxiv2025

Cellular fitness is determined by ribosomal protein S12-mediated release of a truncated Xrp1

Multicellular tissues require continuous optimization to maintain their structure and function by actively eliminating viable, yet unfit cells via a mechanism known as cell competition. During this process, unfit loser cells commonly upregulate the C/EBP family transcription factor Xrp1, which causes their elimination, indicating that intracellular Xrp1 levels determine cellular fitness. However, despite its central role, the mechanism by which Xrp1 is upregulated in unfit cells remains unknown. Here, we show that Xrp1 is upregulated through a previously unrecognized post-transcriptional regulatory mechanism mediated by the ribosomal protein S12 (RpS12). Surprisingly, Xrp1 mRNA is abundantly expressed even in wild-type cells, yet it is not translated. This is due to an upstream open reading frame (uORF) located in the 5UTR of the initial coding exon of Xrp1 mRNA, which inhibits the translation of the main Xrp1 ORF. Intriguingly, in unfit cells, RpS12 causes splicing-mediated skipping of this initial exon, leading to the use of an alternate start codon that generates a short isoform of Xrp1 protein, causing cell death. Notably, protein structural analysis reveals that RpS12 is highly homologous to that of a spliceosomal component SNU13, suggesting the role of RpS12 in directly regulating alternative splicing of Xrp1 mRNA. Our findings thus provide not only a mechanistic insights into how cellular fitness is determined but also how tissue optimization is achieved.

cell biology↗

bioRxiv2025

The protective role of chloroplast NADH dehydrogenase-like complex (NDH) against PSI photoinhibition under chilling stress

Chilling stress induces photosystem I (PSI) photoinhibition in various plants, severely impairing their growth. However, the mechanisms suppressing chilling-induced PSI photoinhibition remain unclear. This study aimed to identify factors preventing PSI photoinhibition by comparing two cucumber cultivars with different susceptibilities to PSI photoinhibition and chilling stress tolerance. In the chilling-sensitive cultivar, partial degradation of the CF1-{gamma} subunit of chloroplast ATPase led to uncoupling of the thylakoid membrane. In addition, electron efflux from Fe-S clusters downstream of PSI was restricted under chilling stress, resulting in highly reduced Fe-S clusters. Notably, this PSI over-reduction in the chilling-sensitive cultivar was observed not only under chilling stress but also under fluctuating light conditions, limited CO2 conditions, and during the transition from darkness to light, suggesting that cyclic electron flow contributes to cultivar differences in PSI photoinhibition. Indeed, the chilling-tolerant cultivar exhibited higher activity of the chloroplast NADH dehydrogenase-like complex (NDH) and suppressed reactive oxygen species (ROS) accumulation during the early stages of chilling stress. In contrast, in the chilling-sensitive cultivar, destabilization of PSI-NDH supercomplex under chilling stress led to the loss of NDH activity, resulting in severe PSI over-reduction. This study provides evidence that NDH acts as a crucial electron sink to prevent PSI photoinhibition and provides new insights into the mechanisms underlying low-temperature stress tolerance.

plant biology↗

bioRxiv2025

PSII photoinhibition as a protective strategy against PSI photoinhibition: Maintaining PSI in an oxidized state by suppressing PSII activity under environmental stress

Photosystem I (PSI) can be photoinhibited by excessive electron flow from Photosystem II (PSII), causing serious growth inhibition due to PSIs limited repair capacity. In contrast, PSII is more susceptible to photoinhibition under stress but has an efficient repair system. Consequently, PSII photoinhibition is considered a protective mechanism that mitigates PSI over-reduction. However, this photoprotective role under environmental stress remains unexplored in intact plants without using mutants or chemical treatments. To address this, we examined the relationship between PSII photoinhibition and susceptibility to PSI photoinhibition under two representative stresses that selectively induce PSI photoinhibition: chilling stress and fluctuating light, using A. thaliana and cucumber. Under chilling stress, A. thaliana exhibited marked PSII photoinhibition and maintained active PSI, whereas cucumber showed insufficient PSII downregulation and suffered PSI photoinhibition. In addition, when fluctuating light treatment was applied to plants with various Fv/Fm (the maximum quantum yield of PSII), plants with reduced Fv/Fm maintained oxidized PSI, and PSI photoinhibition progressed slowly. The susceptibility to PSI photoinhibition under fluctuating light strongly correlated with Fv/Fm, providing clear evidence that PSII photoinhibition protects PSI in vivo. Interestingly, even in plants where P700 remained oxidized during saturation pulses due to PSII photoinhibition, the iron-sulfur clusters (Fe-S clusters) remained reduced. However, the re-oxidation rate of reduced Fe-S clusters was enhanced in PSII-photoinhibited plants, suggesting that charge recombination to P700+ with reduced components on the acceptor side would suppress ROS generation downstream of PSI. This study clarifies how PSII photoinhibition suppresses PSI photoinhibition and prevents ROS-induced damage in wild-type plants under environmental stress.

plant biology↗

bioRxiv2025

Characterization of a crustacean hyperglycemic hormone of the horsehair crab Erimacrus isenbeckii

The horsehair crab Erimacrus isenbeckii is widely distributed from Alaska and the Bering Sea through Southern Sakhalin, and in Japan from the coastal areas of Hokkaido to the Sea of Japan. In the Hokkaido area, although strict resource management has been promoted by setting an allowable catch limit, the catch amount has remained at a low level. While knowledge on larval rearing methods is accumulating in relation to seedling production techniques, information on adult growth and molting is limited, due to a deep-sea species that requires a long period for growth. In decapod crustaceans, the sinus-gland/X-organ complex in the eyestalk ganglion synthesizes and secrets various neuropeptides such as crustacean hyperglycemic hormone (CHH) to regulate the homeostasis of blood glucose levels. In this study, combined isolation of sinus gland peptides by reverse-phase high-performance liquid chromatography (RP-HPLC) and amino acid sequencing, and transcriptome analyses using male and female eyestalk ganglion has been successfully identified EiCHHa. The in vivo assays of EiCHHa using the blue swimming crab Portunus pelagicus revealed EiCHHa has a hyperglycemic effect, as well as CHHs in other decapod crustaceans. Additionally, we successfully demonstrated the sexual differences in the transcriptomic profiles between males and females. Especially, two sinus gland-derived neuropeptides (EiCHHb and a crustacean female sex hormone (EiCFSH)) were isolated as female-biased transcripts, suggesting that both hormones may have female-specific roles such as the development of female characteristics and reproduction.

molecular biology↗

bioRxiv2024

Atlas of Cynomolgus Macaque Hematopoiesis

Self-renewal and differentiation are inherent properties of hematopoietic stem cells (HSCs) that are necessary to support hematopoiesis; however, the underlying mechanisms, especially in human, remain unclear. Here, using the cynomolgus macaque as a surrogate model, we develop a new gating strategy to isolate with high purity transplantable cynomolgus HSCs and generated a single-cell transcriptomic map of cynomolgus HSCs and progenitor cells -- that covering gestational periods previously not analyzed in human. We show that hematopoietic cells from the late-1st to early-3rd trimester fetal liver and late-2nd trimester and thereafter bone marrow have repopulating potential, closely mimicking humans. Unexpectedly however, we found unlike in human, cynomolgus HSCs express CD38 but not CD33, indicating that these cellular counterparts are molecularly distinct. Our transcriptomic analysis reveals the presence of a direct differentiation pathway from HSCs to megakaryocyte lineages, lineage-primed multipotent progenitors and also identified putative HSC surface markers. Taken together, our comprehensive dataset highlights not only the utility of cynomolgus monkeys as model systems to study hematopoiesis but also their potential for translational applications.

cell biology↗

bioRxiv2023

ULK1-regulated AMP sensing by AMPK and its application for the treatment of chronic kidney disease

AMP-activated protein kinase (AMPK) is a central kinase involved in energy homeostasis. Increased intracellular adenosine monophosphate (AMP) levels result in AMPK activation through the binding of AMP to the {gamma}-subunit of AMPK. Recently, we reported that AMP-induced AMPK activation is impaired in the kidneys in chronic kidney disease (CKD) despite an increase in the AMP/ATP ratio. However, the mechanisms by which AMP sensing is disrupted in CKD are unclear. In this study, we identified mechanisms of energy homeostasis in which Unc-51-like kinase 1 (ULK1)-dependent phosphorylation of AMPK{gamma}1 at Ser260/Thr262 promotes AMP sensitivity of AMPK. AMPK activation by AMP was impaired in Ulk1-/- mice despite an increased AMP/ATP ratio. We also demonstrated that MK8722, an allosteric AMPK activator, activates AMPK in the kidneys of a CKD mouse model via a pathway that is independent of AMP sensing. MK8722 treatment significantly attenuates the deterioration of renal function in CKD and is a potential therapeutic option in CKD therapeutics.

molecular biology↗

bioRxiv2022

Epigenetic silencing by SMYD3 represses tumor intrinsic interferon response in HPV-negative squamous cell carcinoma of the head and neck.

Cancers often display immune escape, but the mechanisms and potential for reversibility are incompletely understood. Epigenetic dysregulation has been implicated in the immune escape of various cancer types. We have identified the epigenetic modifier SET and MYND-domain containing protein 3 (SMYD3) as a mediator of immune escape in human papilloma virus (HPV)- negative head and neck squamous cell carcinoma (HNSCC), an aggressive disease with poor prognosis and low response to immunotherapy with pembrolizumab, a programmed-death-1 (PD-1) targeting antibody. SMYD3 loss increased the sensitivity of HNSCC cancer cells to IFN-{beta}, resulting in upregulation of type I IFN response and antigen presentation machinery genes. We found that SMYD3 regulates the transcription of Ubiquitin-Like PHD And RING Finger Domain- Containing Protein 1 (UHRF1), a key epigenetic reader of trimethylated lysine 9 on histone H3 (H3K9me3), which binds to H3K9me3-enriched promoters of key immune-related genes and silences their expression. SMYD3 further maintains the repression of immune-related genes through the deposition of H4K20me3 within the gene body regions of these genes. In an anti-PD-1 immune checkpoint resistant syngeneic mouse model of HPV-negative HNSCC, Smyd3 depletion induced influx of CD8+ T-cells, upregulated PD-L1 and MHC class I molecules, and increased sensitivity to anti-PD-1 therapy. SMYD3 overexpression was associated with decreased CD8 T-cell infiltration in tumor samples from patients with HPV-negative HNSCC, and was associated with poor response to pembrolizumab. Overall, these data highlight a previously unreported function of SMYD3 as a master epigenetic regulator of anti-tumor immune response in HPV-negative HNSCC and provide a rationale for translational approaches combining SMYD3 depletion strategies with checkpoint blockade to overcome anti-PD-1 resistance in this devastating disease.

cancer biology↗

bioRxiv2022

A mouse model of insomnia with sleep apnea

Obstructive sleep apnea (OSA) patients are exposed to nighttime hypoxia during sleep by intermittent airway closure and feel daytime strong sleepiness. Strangely, insomnia co-occur in some OSA patients, which is called co-morbid insomnia and sleep apnea (COMISA). Here, we show activity responses to daytime hypoxia (DHx) in nocturnal mice were comparable to daytime sleepiness and co-occurring nighttime insomnia in COMISA. DHx reduced activity in active phase (AP) and increased following activity in activity ending phase (AEP). This down-and-up activity response (DUR) by DHx was also observed in molecular clock deficient Cry1 and Cry2 double knockout mice (CryDKO) expressing nighttime activity rise under light-dark cycle (LD) and not observed in arrhythmic CryDKO under constant darkness (DD). When daytime timing hypoxia was exposed at transition from LD to DD, about every 6 h down and up and down wavelike activity responses appeared in arrhythmic CryDKO. Results indicate this wavelike response and AP activity overlap and cause DUR in rhythmic mice. DHx increased plasma corticosterone and this increase antagonized AP activity reduction by DHx. DHx reduced forebrain adenosine and morning adenosine inhibition by caffeine induced DUR. Adenosine inhibition by caffeine or istradefylline at transition from LD to DD induced wavelike response in CryDKO. It is possible that wavelike response is damped oscillation because, interestingly, chronic caffeine treatment induced circasemidian and/or circadian activity rhythms in arrhythmic CryDKO. Evening caffeine attenuated DUR by DHx, which suggested adenosine inhibition chronotherapy may improve OSA/COMISA symptoms. Our animal model will be useful to understand COMISA. SignificanceObstructive sleep apnea patients (OSA) are exposed to nighttime hypoxia during sleep. OSA feels daytime strong sleepiness and increases risk of many diseases. Insomnia occurs in not a few OSA, which is called comorbid insomnia and sleep apnea (COMISA). We show here a mouse model of COMISA. In mice, daytime hypoxia exposure induced following down and up activity response (DUR), activity reduction in active phase and increase in activity ending phase, which corresponded to sleepiness and insomnia in COMISA. We found DUR was clock gene independent and might be driven by circasemidian system. Glucocorticoid and forebrain adenosine response were involved in DUR. Caffeine chronotherapy was effective in DUR. Our model may be useful to understand COMISA.

physiology↗

bioRxiv2022

Catecholaminergic cell type-specific expression of Cre recombinase in knock-in transgenic rats generated by the Combi-CRISPR technology

BackgroundCell groups containing catecholamines provide a useful model to study the molecular and cellular mechanisms underlying the morphogenesis, physiology, and pathology of the central nervous system. For this purpose, it is necessary to establish a system to induce catecholaminergic group-specific expression of Cre recombinase. Recently, we introduced a gene cassette encoding 2A peptide fused to Cre recombinase into the site between the C-terminus and translational termination codons of the rat tyrosine hydroxylase (TH) open reading frame by the Combi-CRISPR technology, which is a genomic editing method to enable an efficient knock-in (KI) of long DNA sequence into a target site. However, the expression patterns of the transgene and its function as well as the effect of the mutation on the biochemical and behavioral phenotypes in the KI strains have not been characterized yet. New MethodWe aimed to evaluate the usefulness of TH-Cre KI rats as an experimental model for investigating the structure and function of catecholaminergic neurons in the brain. ResultsWe detected cell type-specific expression of Cre recombinase and site-specific recombination activity in the representative catecholaminergic groups in the TH-Cre KI rat strains. In addition, we measured TH expression level and catecholamine accumulation in the brain regions, and spontaneous locomotion, indicating that catecholamine metabolism and general behavior are apparently normal in these KI rats. ConclusionsTH-Cre KI rat strains produced by the Combi-CRISPR system offer a beneficial model to study the molecular and cellular mechanics for the morphogenesis, physiology, and pathology of catecholamine-containing neurons in the brain.

neuroscience↗

bioRxiv2022

Highly selective transgene expression through double-floxed inverted orientation system by using a unilateral spacer sequence

The double-floxed inverted orientation (DIO) system with adeno-associated viral (AAV) vector provides a beneficial approach to express transgenes in specific cell populations having Cre recombinase. A significant issue with this system is the protection against non-specific expression of transgenes in tissues after vector injection. We here show that Cre-independent recombination in AAV genome carrying the DIO sequence occurs during the production of viral vectors in packaging cells, which results in transgene expression in off-target populations. Introduction of a relatively longer nucleotide sequence between two recognition sites at the unilateral side of the transgene cassette, termed a unilateral spacer sequence (USS), is useful to suppress recombination during the vector production, leading to the protection of non-specific transgene expression with enhanced gene expression selectivity. Our DIO/USS system offers a powerful strategy for highly specific Cre-dependent transgene expression, aiming at various applications for structural and functional analyses of target cell populations.

neuroscience↗

bioRxiv2020

Activity-specificity trade-off gives PI5P4Kβ a nucleotide preference to function as a GTP-sensing kinase.

Most kinases function with ATP. However, contrary to the prevailing dogma, phosphatidylinositol 5-phosphate 4-kinase {beta} (PI5P4K{beta}) utilizes GTP as a primary phosphate donor with a unique binding mode for GTP. Although PI5P4K{beta} is evolved from a primordial ATP-utilizing enzyme, PI4P5K, how PI5P4K{beta} evolutionarily acquired the GTP preference to function as a cellular GTP sensor remains unclear. In this study, we show that the short nucleotide base-recognition motif, TRNVF, is responsible for the GTP binding of PI5P4K{beta}, and also confers onto PI5P4K{beta} an unexpected specificity that extends to inosine triphosphate (ITP) and xanthosine triphosphate (XTP). A mutational study with GTP analogues suggests that the extended specificity is an obligatory consequence to the acquisition of GTP-dependent activity. However, as the cellular concentrations of ITP and XTP are typically negligible, PI5P4K{beta} can still function as a GTP sensor, suggesting that the cellular physiological conditions leave room for the functional evolution of PI5P4K{beta}.

biochemistry↗

bioRxiv2020

A synthetic synaptic organizer protein restores glutamatergic neuronal circuits

Neuronal synapses undergo structural and functional changes throughout life, essential for nervous system physiology. However, these changes may also perturb the excitatory/inhibitory neurotransmission balance and trigger neuropsychiatric and neurological disorders. Molecular tools to restore this balance are highly desirable. Here, we report the design and characterization of CPTX, a synthetic synaptic organizer combining structural elements from cerebellin-1 and neuronal pentraxin-1 to interact with presynaptic neurexins and postsynaptic AMPA-type ionotropic glutamate receptors. CPTX induced the formation of excitatory synapses in vitro and in vivo and restored synaptic functions, motor coordination, spatial and contextual memories, and locomotion in mouse models for cerebellar ataxia, Alzheimers disease and spinal cord injury, respectively. Thus, CPTX represents a prototype for novel structure-guided biologics that can efficiently repair or remodel neuronal circuits. One Sentence SummaryStructural biology information was used to design CPTX, a synthetic protein that induces functional excitatory synapses and restores normal behaviors in mouse models of neurological diseases.

neuroscience↗

bioRxiv2019

Subcellular localization of glypican-5 associates with dynamic cell motility and cell communication of the human mesenchymal stem cell line U3DT

Glypican-5 (GPC5) is a heparan sulfate proteoglycan (HSPG) localized to the plasma membrane. We previously reported that in the human mesenchymal stem cell line UE6E7T3, GPC5 is overexpressed in association with transformation and promotes cell proliferation by acting as a co-receptor for Sonic hedgehog signaling. In this study, we found using an immunofluorescence microscopy that in transformed cells (U3DT), GPC5 localized not only at primary cilia on the cell surface, but also at the leading edge of migrating cells, at the intercellular bridge and blebs during cytokinesis, and in extracellular vesicles. In each subcellular region, GPC5 colocalized with the small GTPase Rab11. These observations suggest that the colocalization of GPC5 with Rab11 is crucial for its function, and that its activity in each subcellular compartment promotes proliferation of U3DT cells. Our findings indicate that GPC5 plays active and essential roles in regulation of U3DT cell proliferation, and provides several insights into the functions of GPC5 that could be elucidated by future studies.

cell biology↗