Biology subjects

Lee, D.

Publications and source records attributed to Lee, D..

At least 109 records · Page 6Linked to original sources

bioRxiv2024

The balance stabilising benefit of social touch: influence of an individual's age and the partner's relative body characteristics

Interpersonal touch (IPT) is a successful strategy to support the stability of and individuals body balance during a multitude of activities in daily life, including physical education and therapy. Despite common practice, however, the influence of an individuals anthropometry and other personal characteristics, such as age, balancing skills, motor experience, and sex as well as interindividual differences in these characteristics between interaction partners on the balance stabilising benefit of social touch is unknown. We assessed an individuals balance stability and change due to IPT provision during single-legged stance in 72 pairs (age range 4 to 63 years) under four sensory conditions: with or without vision in combination with IPT or without. Two participant subgroups were created: one of more vulnerable with low stability and one of more stable, mature participants. Best fitting multiple linear regression models, including moderating variables, for explaining the benefit of IPT in each visual condition indicated that without vision, an individuals benefit of IPT was determined by their balancing skill and the partner-related difference in balancing skill but not by any other factors or partner-related differences. Especially vulnerable individuals improved considerably with IPT when vision was unavailable. With vision complementing IPT, however, an individuals age-dependent motor developmental potential became an additional moderating factor. These findings indicate that the extent to which IPT is benefitting mutual balance stabilisation does not depend on biomechanical factors. Instead, the IPT benefit emerges as a product of both partners sensorimotor capabilities moderated by a persons motor developmental potential when visual feedback could be utilised. We discuss a theoretical framework that accounts for the observed dependencies of the effect of haptic social support on balance control. Significance StatementThis work contributes to a better understanding of the factors influencing the benefit of interpersonal touch for balance stabilization across a wide age range from children to senior adults. It is the first work that investigates the role of both individual factors and relative differences between interaction partners on the amount of benefit of IPT and takes moderating factors such as age-related motor developmental potential, sex, and body mass index into account. Future studies can build on this work when selecting or simulating an optimal partner, especially for individuals with postural instability, for example due to age or sensorimotor deficits.

neuroscience↗

bioRxiv2024

Genotype-immunophenotype relationships in NPM1-mutant AML clonal evolution uncovered by single cell multiomic analysis

Acute myeloid leukemia (AML) is a multi-clonal disease, existing as a milieu of clones with unique but related genotypes as initiating clones acquire subsequent mutations. However, bulk sequencing cannot fully capture AML clonal architecture or the clonal evolution that occurs as patients undergo therapy. To interrogate clonal evolution, we performed simultaneous single cell molecular profiling and immunophenotyping on 43 samples from 32 NPM1-mutant AML patients at different stages of disease. Here we show that diagnosis and relapsed AML samples display similar clonal architecture patterns, but signaling mutations can drive increased clonal diversity specifically at relapse. We uncovered unique genotype-immunophenotype relationships regardless of disease state, suggesting leukemic lineage trajectories can be hard-wired by the mutations present. Analysis of longitudinal samples from patients on therapy identified dynamic clonal, transcriptomic, and immunophenotypic changes. Our studies provide resolved understanding of leukemic clonal evolution and the relationships between genotype and cell state in leukemia biology.

cancer biology↗

bioRxiv2024

CCR2-driven monocyte recruitment is protective against radiotherapy-induced intestinal toxicity

Radiotherapy (RT) is essential in treating abdominal and pelvic cancers but often damages the healthy tissues, particularly the intestines, leading to radiation-induced toxicities with limited treatment options. While the immune system is known to help regulate tissue damage, immune mechanisms involved in RT-induced intestinal toxicity are not fully understood. Following CT-guided localised intestinal irradiation, single-cell RNA sequencing (scRNA-seq) and flow cytometry revealed RT-induced chemokine-dependent recruitment of innate immune cells. Deletion of C-C chemokine receptor (Ccr)1, Ccr2, Ccr3 and Ccr5, blocked recruitment and worsened radiation induced toxicities, suggesting an important role for an innate immune cell population in limiting RT-mediated bowel damage. Furthermore, CCR2-deficient mice showed exacerbated weight loss and intestinal permeability, while the transfer of Ly6C+ monocytes alleviated symptoms. Mechanistically, IL-17 cytokine production by group 3 innate lymphoid cells (ILC3s), a critical factor in maintaining intestinal barrier integrity, was found to be reduced in irradiated CCR2-/-, however the transfer of Ly6C+ monocytes resulted in increased IL-17 levels. These findings demonstrate the critical importance of CCR2-mediated monocyte recruitment in mitigating RT-induced toxicities. One Sentence SummaryCCR2-mediated monocyte recruitment protects against RT-induced intestinal toxicity via IL-17, highlighting a therapeutic target.

immunology↗

bioRxiv2024

Multi-Omics Single-Cell Analysis Reveals Key Regulators of HIV-1 Persistence and Aberrant Host Immune Responses in Early Infection

The clearance of human immunodeficiency virus-1 (HIV-1) remains a significant public health challenge due to impaired cellular immune responses and HIV-1 maintenance during acute infection. However, the genetic and epigenetic changes influencing the immune response on host infected cells remain unclear. Here, this study analyzes HIV-1 infected CD4+ T cells from peripheral blood mononuclear cells from people living with HIV-1 (PLWH) during early infection (<6 months) using single-cell RNA and ATAC sequencing. It is observed that HIV-1 hinders the antiviral response, particularly by interfering with the interferon signaling pathway. Multimodal analysis identifies KLF2 as a key transcription factor in infected CD4+ T cells. Moreover, cells harboring HIV-1 provirus are predominantly identified as Th17 cells, which exhibit elevated KLF2 activity. This suggests an increased susceptibility to HIV-1 infection and a constrained immune response due to the quiescent characteristics of these cells. The finding provides insights into the immune mechanisms and key regulators of HIV-1 maintenance in CD4+ T cells during the early stages of infection.

microbiology↗

bioRxiv2024

TRIM24 directs replicative stress responses to maintain ALT telomeres via chromatin signaling

An inability to replicate the genome can cause replication stress and genome instability. Here, we develop BLOCK-ID, a proteomic method to identify and visualize proteins at stressed replication forks. This approach successfully identified novel mediators of the replication stress response, including the chromatin acetylation reader protein TRIM24. In validating TRIM24 function, we uncovered its crucial role in coordinating Alternative Lengthening of Telomeres (ALT), a cancer-specific telomere extension pathway involving replication stress. Our data reveal that TRIM24 is directed to telomeres via a p300/CBP-dependent acetylation chromatin signaling cascade, where it organizes ALT-associated PML bodies (APBs) to promote telomere DNA synthesis. Strikingly, we demonstrate that when artificially tethered at telomeres, TRIM24 can stimulate new telomere DNA synthesis in a SUMO-dependent manner, independently of p300/CBP or PML-dependent APBs. Thus, this study identifies a TRIM24 chromatin signaling pathway required for ALT telomere maintenance.

cell biology↗

bioRxiv2024

Hippocampal-cortical interactions in the consolidation of social memory

Episodic memories are initially encoded in the hippocampus and subsequently undergo systems consolidation into the neocortex. The nature of memory stored in the hippocampus and neocortex also differs, with the cortex shown to encode memories in more generalized forms. Although several brain regions are known to encode social information, the specific cortical regions and circuits involved in the consolidation of social memory and the nature of the information encoded in the cortex remain unclear. Using in vivo Ca2+ imaging and optogenetic manipulations, we found that infralimbic (IL) neurons projecting to the nucleus accumbens shell (IL[->]NAcSh) neurons store consolidated social memory. Inactivating IL[->]NAcSh neurons that responded to a familiar conspecific impaired the recognition of other familiar mice including littermates, demonstrating that these neuronal activities support social familiarity. Furthermore, inactivating hippocampal ventral CA1 neurons projecting to the IL disrupted the consolidation of memory for newly familiarized mice while sparing the recognition of littermates. These findings demonstrate the critical role of hippocampal-cortical interactions in the consolidation of social memory.

animal behavior and cognition↗

bioRxiv2024

Tuning Insulin Receptor Signaling Using De Novo Designed Agonists

Binding of insulin to the insulin receptor (IR) induces conformational changes in the extracellular portion of the receptor that lead to activation of the intracellular kinase domain and the AKT and MAPK pathways, and downstream modulation of glucose metabolism and cell proliferation. We reasoned that designed agonists that induce different conformational changes in the receptor might induce different downstream responses, which could be useful both therapeutically and to shed light on how extracellular conformation is coupled to intracellular signaling. We used de novo protein design to first generate binders to individual IR extracellular domains, and then to fuse these together in different orientations and with different conformational flexibility. We describe a series of synthetic agonists that signal through the IR that differ from insulin and from each other in the induction of receptor autophosphorylation, MAPK activation, intracellular trafficking, and cell proliferation. We identify designs that are more potent than insulin causing much longer lasting reductions in glucose levels, and that retain signaling activity on disease-causing receptor mutants that do not respond to insulin. These results inform our understanding of how changes in receptor conformation and dynamics are transmitted to downstream signaling, and our synthetic agonists have considerable therapeutic potential for diabetes and severe insulin resistance syndromes. HighlightsO_LIComputational design yielded super agonists, partial agonists, and antagonists of IR. C_LIO_LIDe novo agonists induce a distinct IR active conformation. C_LIO_LIDesigned agonists tune IR signaling by modulating conformational dynamics of activated IR. C_LIO_LIDesigned agonists are more potent than insulin, reducing glucose levels longer and activating disease-causing IR mutants. C_LI

bioengineering↗

bioRxiv2024

Adaptation of retinal discriminability to natural scenes

Sensory systems discriminate between stimuli to direct behavioral choices, a process governed by two distinct properties - neural sensitivity to specific stimuli, and neural variability that importantly includes correlations between neurons. Two questions that have received extensive investigation and debate are whether visual systems are optimized for natural scenes, and whether correlated neural variability contributes to this optimization. However, the lack of sufficient computational models has made these questions inaccessible in the context of the normal function of the visual system, which is to discriminate between natural stimuli. Here we take a direct approach to analyze discriminability under natural scenes for a population of salamander retinal ganglion cells using a model of the retinal neural code that captures both sensitivity and variability. Using methods of information geometry and generative machine learning, we analyzed the manifolds of natural stimuli and neural responses, finding that discriminability in the ganglion cell population adapts to enhance information transmission about natural scenes, in particular about localized motion. Contrary to previous proposals, correlated noise reduces information transmission and arises simply as a natural consequence of the shared circuitry that generates changing spatiotemporal visual sensitivity. These results address a long-standing debate as to the role of retinal correlations in the encoding of natural stimuli and reveal how the highly nonlinear receptive fields of the retina adapt dynamically to increase information transmission under natural scenes by performing the important ethological function of local motion discrimination.

neuroscience↗

bioRxiv2024

Cell-type-specific mapping of enhancers and target genes from single-cell multimodal data

Mapping enhancers and target genes in disease-related cell types has provided critical insights into the functional mechanisms of genetic variants identified by genomewide association studies (GWAS). However, most existing analyses rely on bulk data or cultured cell lines, which may fail to identify cell-type-specific enhancers and target genes. Recently, single-cell multimodal data measuring both gene expression and chromatin accessibility within the same cells have enabled the inference of enhancer-gene pairs in a cell-type-specific and context-specific manner. However, this task is challenged by the datas high sparsity, sequencing depth variation, and the computational burden of analyzing a large number of enhancer-gene pairs. To address these challenges, we propose scMultiMap, a statistical method that infers enhancer-gene association from sparse multimodal counts using a joint latent-variable model. It adjusts for technical confounding, permits fast moment-based estimation and provides analytically derived p-values. In systematic analyses of blood and brain data, scMultiMap shows appropriate type I error control, high statistical power with greater reproducibility across independent datasets and stronger consistency with orthogonal data modalities. Meanwhile, its computational cost is less than 1% of existing methods. When applied to single-cell multimodal data from postmortem brain samples from Alzheimers disease (AD) patients and controls, scMultiMap gave the highest heritability enrichment in microglia and revealed new insights into the regulatory mechanisms of AD GWAS variants in microglia.

bioinformatics↗

bioRxiv2024

Engineered Receptors for Soluble Cell-to-Cell Communication

Despite recent advances in mammalian synthetic biology, there remains a lack of modular synthetic receptors that can robustly respond to soluble ligands and in turn activate cellular functions. Such receptors would have extensive clinical potential to regulate the activity of engineered therapeutic cells, but to date only receptors against cell surface targets have approached clinical translation. To address this gap, we developed a receptor architecture based on synNotch, called SyNthetic Intramembrane Proteolysis Receptor (SNIPR), that has the added ability to be activated by soluble ligands, both natural and synthetic, with remarkably low baseline activity and high fold activation. SNIPRs are able to access an endocytic, pH-dependent cleavage mechanism to achieve soluble ligand sensing, in addition to employing a canonical-like pathway for detecting surface-bound ligands. We demonstrate the therapeutic capabilities of the receptor platform by localizing the activity of CAR T-cells to solid tumors where soluble disease-associated factors are expressed, bypassing the major hurdle of on-target off-tumor toxicity in bystander organs. We further applied the SNIPR platform to engineer fully synthetic signaling networks between cells orthogonal to natural signaling pathways, expanding the scope of synthetic biology. Our design framework enables cellular communication and environmental interactions, extending the capabilities of synthetic cellular networking in clinical and research contexts.

bioengineering↗

bioRxiv2024

Accurate antibody loop structure prediction enables zero-shot design of target-specific antibodies

Protein loops, characterized by their versatile structures with varying sizes and shapes, can recognize a wide range of targets with high specificity and affinity. The variable loops of the antibody complementarity-determining region (CDR) are particularly crucial for immune responses and therapeutic applications due to their effective target recognition capabilities. Accurate structure prediction of these antibody loops is essential for the efficient in silico design of target-binding antibodies for therapeutic or industrial use. However, predicting antibody loop structures is challenging due to the lack of evolutionary information from related proteins. Thus, a successful ab initio structure prediction method, which operates without structural templates or related sequences, is crucial for the effective design of antibody loop-mediated interactions. This study demonstrates that highly accurate antibody loop structure prediction enables the effective zero-shot design of target-binding antibody loops. The performance of loop design has been shown to depend on the accuracy of ab initio loop structure prediction, as tested with two versions of our design model. The high affinity, diversity, novelty, and specificity of the antibody loops designed with these new methods were validated experimentally on four target proteins.

bioengineering↗

bioRxiv2024

Spatial variability of agricultural soil carbon dioxide and nitrous oxide fluxes: characterization and recommendations from spatially high-resolution, multi-year dataset

Mitigating agricultural soil greenhouse gas (GHG) emissions can contribute to meeting the global climate goals. High spatial and temporal resolution, large-scale, and multi-year data are necessary to characterize and predict spatial patterns of soil GHG fluxes to establish well-informed mitigation strategies, but not many of such datasets are currently available. To address this gap in data we collected two years of high spatial resolution (7.4 sampling points ha-1 over 2.0 to 5.4 ha area) in-season soil carbon dioxide (CO2) and nitrous oxide (N2O) fluxes from three commercial sites in central Illinois, one conventionally managed continuous corn and two under conservation practices in corn-soybean rotations typical of the region. At the field-scale, the spatial variability of CO2 was comparable across sites, years, and management practices, but N2O was on average 77% more spatially variable in the conventionally managed site. Analysis of N2O hotspots revealed that although they represent a similar proportion of the sampling areas across sites (conventional: 12%; conservation: 13%), hotspot contribution to field-wide emission was greater in the conventional site than in the conservation sites (conventional: 51%; conservation: 34%). Also, the spatial patterns, especially hotspot locations, of both gases were inter-annually inconsistent, with hotspots rarely occurring in the same location. Overall, our result indicated that traditional field-scale monitoring with gas chambers may not be the optimal approach to detect GHG hotspots in row crop systems, due to the unpredictable spatial heterogeneity of management practices. Still, our sensitivity analysis on the dataset demonstrated that sampling at a spatial resolution of 1.6 and 5.6 points ha-1 can provide reliable (< 25% error) estimates of field-scale soil CO2 and N2O fluxes, respectively.

biochemistry↗

bioRxiv2024

Wide-field bond-selective fluorescence imaging: from single-molecule to cellular imaging beyond video-rate

Wide-field (WF) imaging is pivotal for observing dynamic biological events. While WF chemical microscopy offers high molecular specificity, it lacks the sensitivity for single-molecule detection. In contrast, WF fluorescence microscopy provides live-cell dynamic mapping but fails to leverage the rich chemical information necessary for functional interpretations. To address these limitations, we introduce Wide-Field Bond-selective Fluorescence-detected Infrared-Excited (WF-BonFIRE) spectro-microscopy. This technique combines rationally optimized imaging speed and field-of-view (FOV) to achieve single-molecule sensitivity with bond-selective contrast. WF-BonFIRE outperforms its point-scanning counterpart, enhancing frame acquisition up to 10,000 times. We demonstrate WF-BonFIREs capabilities in imaging cells, astrocytes, and live neurons, capturing single FOVs up to 50 {micro}m x 50 {micro}m, with further expansion via multi-FOV mosaicking. Additionally, we have implemented a temporal-delay modulation scheme that allows real-time kilohertz imaging speeds up to 1500 Hz. This enables millisecond temporal resolution while monitoring random motion of live Escherichia coli. Overall, WF-BonFIRE significantly broadens the possibilities for chemical imaging, enabling high-speed observations at unparalleled sensitivity levels. One-Sentence SummaryWide-field bond-selective fluorescence imaging pushes chemical-sensitive microscopy platform into a new regime, achieving single-molecule sensitivity and speeds up to kilohertz.

biophysics↗

bioRxiv2024

Control of innate olfactory valence by segregated cortical amygdala circuits

Animals exhibit innate behaviors that are stereotyped responses to specific evolutionarily relevant stimuli in the absence of prior learning or experience. These behaviors can be reduced to an axis of valence, whereby specific odors evoke approach or avoidance responses. The posterolateral cortical amygdala (plCoA) mediates innate attraction and aversion to odor. However, little is known about how this brain area gives rise to behaviors of opposing motivational valence. Here, we sought to define the circuit features of plCoA that give rise to innate attraction and aversion to odor. We characterized the physiology, gene expression, and projections of this structure, identifying a divergent, topographic organization that selectively controls innate attraction and avoidance to odor. First, we examined odor-evoked responses in these areas and found sparse encoding of odor identity, but not valence. We next considered a topographic organization and found that optogenetic stimulation of the anterior and posterior domains of plCoA elicits attraction and avoidance, respectively, suggesting a functional axis for valence. Using single cell and spatial RNA sequencing, we identified the molecular cell types in plCoA, revealing an anteroposterior gradient in cell types, whereby anterior glutamatergic neurons preferentially express VGluT2 and posterior neurons express VGluT1. Activation of these respective cell types recapitulates appetitive and aversive behaviors, and chemogenetic inhibition reveals partial necessity for responses to innate appetitive or aversive odors. Finally, we identified topographically organized circuits defined by projections, whereby anterior neurons preferentially project to medial amygdala, and posterior neurons preferentially project to nucleus accumbens, which are respectively sufficient and necessary for innate attraction and aversion. Together, these data advance our understanding of how the olfactory system generates stereotypic, hardwired attraction and avoidance, and supports a model whereby distinct, topographically distributed plCoA populations direct innate olfactory responses by signaling to divergent valence-specific targets, linking upstream olfactory identity to downstream valence behaviors, through a population code. This suggests a novel amygdala circuit motif in which valence encoding is represented not by the firing properties of individual neurons, but by population level identity encoding that is routed through divergent targets to mediate distinct behaviors of opposing appetitive and aversive responses.

neuroscience↗

bioRxiv2024

De Novo Design of Integrin α5β1 Modulating Proteins for Regenerative Medicine

Integrin 5{beta}1 is crucial for cell attachment and migration in development and tissue regeneration, and 5{beta}1 binding proteins could have considerable utility in regenerative medicine and next-generation therapeutics. We use computational protein design to create de novo 5{beta}1-specific modulating miniprotein binders, called NeoNectins, that bind to and stabilize the open state of 5{beta}1. When immobilized onto titanium surfaces and throughout 3D hydrogels, the NeoNectins outperform native fibronectin and RGD peptide in enhancing cell attachment and spreading, and NeoNectin-grafted titanium implants outperformed fibronectin and RGD-grafted implants in animal models in promoting tissue integration and bone growth. NeoNectins should be broadly applicable for tissue engineering and biomedicine. One-Sentence SummaryA de novo-designed fibronectin substitute, NeoNectin, is specific for integrin 5{beta}1 and can be incorporated into biomaterials for regenerative medicine.

bioengineering↗

bioRxiv2024

Multispectral live-cell imaging with uncompromised spatiotemporal resolution

Multispectral imaging is an established method to extend the number of colours usable in fluorescence imaging beyond the typical limit of three or four, but standard approaches are poorly suited to live-cell imaging. We introduce an approach for multispectral imaging in live cells, comprising an iterative spectral unmixing algorithm and eight channel camera-based image acquisition hardware. This enables the accurate unmixing of low signal-to-noise ratio datasets, typical of live-cell imaging, captured at video rates. We use this approach on a commercial spinning disk confocal microscope and a home-built oblique plane light sheet microscope to image 1-7 spectrally distinct fluorophore species simultaneously, using both fluorescent protein fusions and small-molecule dyes. We further use de novo designed protein-binding proteins (minibinders), labelled with organic fluorophores, and use these in combination with our multispectral imaging approach to study the endosomal trafficking of cell-surface receptors at endogenous levels.

biophysics↗

bioRxiv2024

Quantum-classical hybrid approach for codon optimizationand its practical applications

Codon optimization is crucial for gene expression in heterologous hosts with varying genetic codes and codon usage, potentially resulting in enhanced protein expression and stability. Traditionally, the codon optimization problem has been solved using classical numerical techniques; however, with recent advancements, quantum algorithms deployed on quantum computers have been adopted for this purpose. This study proposes a codon sequence search protocol tailored to host preferences. Specifically, codon optimization is formulated as a constrained quadratic binary problem and solved using a quantum-classical hybrid approach, integrating quantum annealing with the Lagrange multiplier method. The proposed methodology is then applied to two real-world scenarios: optimizing the codon sequence of the severe respiratory syndrome coronavirus 2 spike protein in human hosts and insulin in Escherichia coli (E. coli) hosts. Finally, evaluations of several biological metrics demonstrate the effectiveness of our protocol, offering insights into the codon usage patterns governing translational efficiency and adaptation to the genetic code preferences of the host organisms.

bioinformatics↗

bioRxiv2024

Endogenous tau released from human ReNCell VM cultures by neuronal activity is phosphorylated at multiple sites

Tau is an intracellular protein but also known to be released into the extracellular fluid. Tau release mechanisms have drawn intense attention as these are known to play a key role in Alzheimers disease (AD) pathology. However, tau can also be released under physiological conditions although its physiological function and release mechanisms have been poorly characterized, especially in human neuronal cells. We investigated endogenous tau release in ReNCell VM, a human neuroprogenitor cell line, under physiological conditions and found that tau is spontaneously released from cells. To study activity-dependent release of endogenous tau, human ReNCell VM culture was stimulated by 100M AMPA or 50mM KCl for one-hour, tau was actively released to the culture medium. The released tau was highly phosphorylated at nine phosphorylation sites (pSites) detected by phospho-specific tau antibodies including AT270 (T175/T181), AT8 (S202/T205), AT100 (T212/S214), AT180 (T231), and PHF-1 (S396/S404), showing that these pSites are important for activity-dependent tau release from human ReNCell VM. Intracellular tau showed various phosphorylation status across these sites, with AT270 and PHF-1 highly phosphorylated while AT8 and AT180 were minimally phosphorylated, suggesting that AT8 and AT180 pSites exhibit a propensity for secretion rather than being retained intracellularly. This activity-dependent tau release was significantly decreased by inhibition of GSK-3{beta}, demonstrating that GSK3{beta}-dependent phosphorylation of tau plays an important role in its release by neuronal activity. In this study, we showed that ReNCell VM serves as a valuable model for studying endogenous physiological tau release. Further, ReNCell model can be also used to study pathological release of human tau that will contribute to our understanding of the progression of AD and related dementias. HighlightsO_LIActivity-dependent release of endogenous human tau from human ReNCell VM cultures occurs under physiological conditions. C_LIO_LIReleased human tau is phosphorylated at nine sites (pSites) in the proline-rich domain and the C-terminal domain detected by AT270 (T175/T181), AT8 (S202/T205), AT100 (T212/S214), AT180 (T231), and PHF-1 (S396/S404) tau antibodies, strongly suggesting that these pSites are important for activity-dependent tau release from human ReNCell VM. C_LIO_LIIn contrast, intracellular human tau proteins have different phosphorylation status among these nine pSites: AT270 and PHF-1 pSites are highly phosphorylated, but AT8 and AT180 are weakly phosphorylated, suggesting AT8 and AT180 pSites are release-sensitive phosphorylation motifs. C_LIO_LIActivity-dependent release of endogenous human tau is decreased by a tau kinase GSK-3{beta} inhibitor SB 216763, indicating that GSK-3{beta}-dependent phosphorylation plays an important role in activity-dependent tau release. C_LIO_LIThe human ReNCell culture is an excellent model system to study mechanisms underlying physiological release of endogenous tau. C_LI

neuroscience↗