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Biology subjects

Luan, L.

Publications and source records attributed to Luan, L..

17 recordsLinked to original sources

Learning induces activation-mechanism--dependent neural plasticity in an intracortical microstimulation task

Electrical microstimulation provides high-resolution control of neural circuits for causal studies and restoration of impaired functions, yet how responses to artificial activation evolve with learning remains unclear. Here, we deploy a detection task and pair ultraflexible electrodes for stable intracortical microstimulation (ICMS) with longitudinal imaging and recordings to track single-cell and population responses across weeks of learning. Detection thresholds decreased with learning, indicating plasticity. Chronic imaging showed that stimulus-evoked recruitment expanded at a fixed current, while a consistent number of neurons continued to underlie behavioral responses. A subset of learning-sensitive cells enhanced modulation and reduced latency. Electrophysiological recordings further distinguished two forms of adaptation: directly activated, pulse-locked neurons strengthened their excitability, whereas polysynaptically recruited neurons expanded in number and were predictive of behavioral outcomes. These results show that learning in an ICMS task reshapes cortical circuits through activation-mechanism-dependent plasticity, underscoring the need for stimulation paradigms that adapt to both cell-intrinsic and network dynamics. One-sentence summaryLearning alters neural responses to ICMS, revealing differences between direct activation and network activity linked to behavior.

neuroscience↗

A multimodal perturbation atlas defines the phenotypic resolution of cellular morphology.

Modeling cellular behavior requires measurements that capture how cells evolve across time, environments, and interventions. Microscopy is uniquely suited to this goal: it is non-destructive and can be applied to living cells in their native context. Yet its phenotypic resolving power remains incompletely characterized relative to molecular assays. Here, we present a multimodal perturbation atlas of 1,000 pooled CRISPR knockouts in A549 cells, profiled by fluorescence microscopy (42 live, 13 fixed markers), label-free quantitative phase imaging of the same live cells (at single timepoints), and single-cell RNA sequencing (scRNA-seq). We develop deep learning frameworks to interpret the rich cell-biological signatures in these ~65M single-cell profiles. At matched reagent cost, phase imaging exceeds the phenotypic resolution of both fluorescence imaging and scRNA-seq, and more reliably recovers higher-order pathway organization. These results establish intrinsic morphology as a high-precision readout of cellular state, and lay a foundation for live-cell profiling of phenotypic trajectories.

systems biology↗

High-channel-count neural recording and stimulation platform with 5,376 simultaneous recording channels

Advancing neural interfaces requires large-scale, high-density recording technologies capable of capturing full-spectrum neural activity across cortical and subcortical regions. Here, we present a scalable approach to integrate neural electrodes with advanced application-specific integrated circuits (ASICs). Specifically, we custom-designed an ASIC with 5,376 simultaneous channels, each sampling at 20 kS/s and enabling >1.3 Gb/s total data streaming throughput. The ASIC incorporates in-pixel amplification, time-division multiplexed ADCs, and on-chip stimulation capabilities, ensuring precise signal acquisition with minimal power consumption while maintaining a low noise level of 5.5 {micro}Vrms. We further developed an interconnect strategy using gold bump bonding, which allows for high-density integration of the flexible probe and rigid chip. We demonstrate the capacity of this platform through the integration with a flexible ECoG array. The resulting device allows for the high-resolution mapping of in vivo field potentials on the cortical surfaces of rat brains, supported by the precise localization of evoked sensory activities. These results prove an effective approach towards highly integrated neural interfaces with applications in brain-computer interfaces, neuroprosthetics, and large-scale functional brain mapping.

bioengineering↗

Temporal coding carries more stable cortical visual representations than firing rate over time

The brains ability to stably represent recurring visual scenes is crucial for behavior. Previous studies have used slow dynamic (1-5 seconds) rate code measurements to study visual tuning, revealing varying degrees of gradual activity changes over time or "representational drifts." However, it remains unclear if there is an underlying neural code that maintains the encoding of information stable over time. In this study, we extracted structures in fast (tens of milliseconds) temporal responses and explored the role of such temporal codes in supporting the stability of visual representations. We tracked the spiking activity of the same visual cortical populations in male mice for 15 consecutive days using custom-developed, large-scale, ultraflexible electrode arrays. Across various types of stimuli, we found that neurons exhibited varying degrees of day-to-day stability in their firing rate-based tuning. The across day stability correlated with tuning reliability. Notably, accounting for spiking temporal dynamics increased single neuron tuning stability, especially for less reliable neurons. Temporal coding further improved population representation discriminability and decoding accuracy. The stability of temporal codes was more correlated with network functional connectivity than rate coding. These results show that temporal coding is crucial for stably encoding sensory stimuli, suggesting its significant role in ensuring consistent sensory experiences.

neuroscience↗

A synthetic chlorophagy receptor promotes plant fitness by mediating chloroplast microautophagy

Chloroplasts are major photosynthetic and protein-containing organelles in green plants and algae. Unwanted chloroplast proteins and entire chloroplasts are cleared through various degradation pathways including autophagy. Nevertheless, canonical chlorophagy receptors remain unidentified, and whether and to what extent chlorophagy can be enhanced to benefect the plants remain unknown. Here we designed and validated a synthetic chloroplast autophagy receptor using biochemical, genetical, and imaging approaches. This synthetic receptor, LIR-SNT-BFP, was constructed by fusing a fragment containing the LC3-interacting region (LIR) of the selective autophagy receptor NBR1 and the N-terminal amphipathic -helix of the chloroplast outer envelope protein SFR2. The fusion protein LIR-SNT-BFP coated the chloroplast and attracted ATG8a in planta. Upon induction, the synthetic receptor elicited vacuole-mediated microautophagy of entire chloroplasts independent of the ATG8 conjugation machinery proteins ATG5 or ATG7. Meanwhile, it induced chloroplast division; however the induced chlorophagy was independent of PDV2. Notably, moderate chlorophagy improves rosette growth, but excessive levels are detrimental. Furthermore, induced chlorophagy partially protects against herbicide-induced leaf chlorosis. This study demonstrates controlled chloroplast degradation using a synthetic chlorophagy receptor.

plant biology↗

DMT-induced shifts in criticality correlate with ego-dissolution

Psychedelics profoundly alter subjective experience and brain dynamics. Brain oscillations express signatures of near-critical dynamics, relevant for healthy function. Alterations in the proximity to criticality have been suggested to underlie the experiential and neurological effects of psychedelics. Here, we investigate the effects of a psychedelic substance (DMT) on the criticality of brain oscillations, and in relation to subjective experience. We find that DMT shifts the dynamics of brain oscillations away from criticality in alpha and adjacent frequency bands. In this context, entropy is increased while complexity is reduced. We find that the criticality shifts observed in alpha and theta bands correlate with the intensity ratings of ego-dissolution, a hallmark of psychedelic experience. Finally, using a recently developed metric, the functional excitatory-inhibitory ratio, we find that the DMT-induced criticality shift in brain oscillations is towards subcritical regimes. These findings have major implications for the understanding of psychedelic mechanisms of action in the human brain and for the neurological basis of altered states of consciousness.

neuroscience↗

Inhibition of SQSTM1/p62 oligomerization and Keap1 sequestration by the Cullin-3 adaptor SHKBP1

SQSTM1/p62 is a master regulator of the autophagic and ubiquitination pathways of protein degradation and the antioxidant response. p62 functions in these pathways via reversible assembly and sequestration of additional factors into cytoplasmic phase-separated structures termed p62 bodies. The physiological roles of p62 in these various pathways depends on numerous mechanisms for regulating p62 body formation and dynamics that are incompletely understood. Here, we identify a new mechanism for regulation of p62 oligomerization and incorporation into p62 bodies by SHKBP1, a Cullin-3 E3 ubiquitin ligase adaptor, that is independent of its potential functions in ubiquitination. We map a SHKBP1-p62 protein-protein interaction outside of p62 bodies that limits p62 assembly into p62 bodies and affects the antioxidant response by preventing sequestration and degradation of Keap1. These studies provide a non-ubiquitination-based mechanism for an E3 ligase adaptor in regulating p62 phase separation and cellular responses to oxidative stress.

cell biology↗

Water use efficiency of Cupressus gigantea is higher in mountain slope habitats compared to riverside habitats

Cupressus gigantea is an endemic species of the Yarlung Zangbo River in Tibet, China, and is designated as a national key protected wild plant. It is only distributed in the narrow area along Yarlung Zangbo River from Jiacha County to Danniang village Milin City in various habitats such as the waterline along the Yarlung Zangbo River, terraces, and mountain slopes. In this study, we utilized stable isotope technology ({delta}D, {delta}18O, and {delta}13C), combined with the IsoSource model, to quantitatively determined the sources of water absorption and the water use efficiency of C. gigantea in various habitats, including riverside area, terraces and mountain slopes. The results showed that the content of soil total nitrogen, soil nitrate nitrogen, soil ammonium nitrogen, and soil organic carbon are significantly higher on mountain slope than in the other two habitats, while the total carbon content, water content, and total nitrogen content of C. gigantea leaves are significantly lower on mountain slope than in the other two habitats (P<0.05). The water use efficiency (WUE) of C. gigantea in the mountain slope habitats is 107.8 mol mol-1, which is significantly higher than that of other habitats (P<0.05), with the WUE of C. gigantea in the riverside habitat being the lowest at 35.7 mol mol-1. C. gigantea distributed on the mountain slope and terrace uses soil water as the main water source. During the normal rainy season, the proportion of soil water utilization is the highest, accounting for more than 60%; in the dry season, the utilization of river water and groundwater increases, while soil water utilization decreases. C. gigantea distributed along the riverside mainly absorbs water from the Yarlung Zangbo River, about 63%. In August when the rainfall increases in the rainy season, the use of soil moisture increases, and in the dry season, river water becomes the main water source. This indicates that with the alternation of rainy and dry seasons, the water absorption sources and strategies of C.gigantea adjusted accordingly. The research results provide an important scientific basis for the protection and management measures of the C. gigantea population.

ecology↗

Neuroprotective Mechanism of Ribisin A on H 2 O 2 -induced PC12 cell injury model

Ribisin A has been shown to have neurotrophic activity. The aim of this study was to evaluate the neuroprotective effect of Ribisin A on injured PC12 cells and elucidate its mechanism. In this project, PC12 cells were induced by H2O2 to establish an injury model. After treatment with Ribisin A, the neuroprotective mechanism of Ribisin A was investigated by methyl tetrazolium (MTT) assay, Enzyme-linked immunosorbent assay (ELISA), flow cytometric analysis, fluorescent probe analysis, and western blot. We found that Ribisin A decreased the rate of lactate dehydrogenase (LDH) release, increased cellular superoxide dismutase (SOD) activity, decreased the levels of tumor necrosis factor- (TNF-), interleukin-6 (IL-6), Ca2+ expression and reactive oxygen species (ROS). Moreover, Ribisin A significantly increased mitochondrial membrane potential (MMP) and inhibited apoptosis of PC12 cells. Meanwhile, Ribisin A activated the phosphorylation of ERK1/2 and its downstream molecule CREB by upregulating the expression of Trk A and Trk B, the upstream molecules of the ERK signaling pathway. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=110 SRC="FIGDIR/small/559840v1_ufig1.gif" ALT="Figure 1"> View larger version (21K): org.highwire.dtl.DTLVardef@ff9604org.highwire.dtl.DTLVardef@1d1ee75org.highwire.dtl.DTLVardef@17e4441org.highwire.dtl.DTLVardef@b347c4_HPS_FORMAT_FIGEXP M_FIG C_FIG

pharmacology and toxicology↗

Low-threshold, high-resolution, chronically stable intracortical microstimulation by ultraflexible electrodes

Intracortical microstimulation (ICMS) enables applications ranging from neuroprosthetics to causal circuit manipulations. However, the resolution, efficacy, and chronic stability of neuromodulation is often compromised by the adverse tissue responses to the indwelling electrodes. Here we engineer ultraflexible stim-Nanoelectronic Threads (StimNETs) and demonstrate low activation threshold, high resolution, and chronically stable ICMS in awake, behaving mouse models. In vivo two-photon imaging reveals that StimNETs remain seamlessly integrated with the nervous tissue throughout chronic stimulation periods and elicit stable, focal neuronal activation at low currents of 2 A. Importantly, StimNETs evoke longitudinally stable behavioral responses for over eight months at markedly low charge injection of 0.25 nC/phase. Quantified histological analysis show that chronic ICMS by StimNETs induce no neuronal degeneration or glial scarring. These results suggest that tissue-integrated electrodes provide a path for robust, long-lasting, spatially-selective neuromodulation at low currents which lessen risks of tissue damage or exacerbation of off-target side-effects.

neuroscience↗

Imaging interorganelle phospholipid transport by extended synaptotagmins using bioorthogonally tagged lipids

The proper distribution of lipids within organelle membranes requires rapid, interorganelle lipid transport, much of which occurs at membrane contact sites and is mediated by lipid transfer proteins (LTPs). Our current understanding of LTP mechanism and function is based largely on structural studies and in vitro reconstitution. Existing cellular assays for LTP function use indirect readouts, and it remains an open question as to whether substrate specificity and transport kinetics established in vitro are similar in cellular settings. Here, we harness bioorthogonal chemistry to develop tools for direct visualization of interorganelle transport of phospholipids between the plasma membrane (PM) and the endoplasmic reticulum (ER). Unnatural fluorescent phospholipid analogs generated by the transphosphatidylation activity of phospholipase D (PLD) at the PM are rapidly transported to the ER dependent in large part upon extended synaptotagmins (E-Syts), a family of LTPs at ER-PM contact sites. Ectopic expression of an artificial E-Syt-based tether at ER-mitochondria contact sites results in fluorescent phospholipid accumulation in mitochondria. Finally, in vitro reconstitution assays demonstrate that the fluorescent lipids are bona fide E-Syt substrates. Thus, fluorescent lipids generated in situ via PLD activity and bioorthogonal chemical tagging can enable direct visualization of the activity of LTPs that mediate bulk phospholipid transport at ER-PM contact sites.

cell biology↗

MyD88-Dependent Signaling Drives Toll-Like Receptor-Induced Trained Immunity in Macrophages

Immunocompromised populations are highly vulnerable to developing life-threatening infections. Strategies to protect patients with weak immune responses are urgently needed. Employing trained immunity, whereby innate leukocytes undergo reprogramming upon exposure to a microbial product and respond more robustly to subsequent infection, is a promising approach. Previously, we demonstrated that the TLR4 agonist monophosphoryl lipid A (MPLA) induces trained immunity and confers broad resistance to infection. TLR4 signals through both MyD88- and TRIF-dependent cascades, but the relative contribution of each pathway to induction of trained immunity is unknown. Here, we show that MPLA-induced resistance to Staphylococcus aureus infection is lost in MyD88-KO, but not TRIF-KO, mice. The MyD88-activating agonist CpG (TLR9 agonist), but not TRIF-activating Poly I:C (TLR3 agonist), protects against infection in a macrophage-dependent manner. MPLA- and CpG-induced augmentation of macrophage metabolism and antimicrobial functions is blunted in MyD88-, but not TRIF-KO, macrophages. Augmentation of antimicrobial functions occurs in parallel to metabolic reprogramming and is dependent, in part, on mTOR activation. Splenic macrophages from CpG-treated mice confirmed that TLR/MyD88-induced reprogramming occurs in vivo. TLR/MyD88-triggered metabolic and functional reprogramming was reproduced in human monocyte-derived macrophages. These data show that MyD88-dependent signaling is critical in TLR-mediated trained immunity.

immunology↗

ATG9 vesicles comprise the seed membrane of mammalian autophagosomes

During autophagosome biogenesis, the incorporation of transmembrane proteins into the expanding phagophore is not readily observed. In addition, the membrane surface area of the organelle expands rapidly, while the volume of the autophagosome is kept low. Several recent studies have suggested a model of membrane expansion that explains how these attributes are maintained. The autophagosome expands predominantly through the direct protein-mediated transfer of lipids through the lipid transfer protein ATG2. As these lipids are only introduced into the cytoplasmic-facing leaflet of the expanding phagophore, full membrane growth also requires lipid scramblase activity. ATG9 has been demonstrated to harbor scramblase activity and is essential to autophagosome formation, however if and when it is integrated into mammalian autophagosomes remains unclear. Here we show that in the absence of lipid transport, ATG9 vesicles are already fully competent to collect proteins normally found on mature autophagosomes, including LC3-II. Further, through the novel use of styrene-maleic acid lipid particles as a nanoscale interrogation of protein organization on intact membranes, we show that ATG9 is fully integrated in the same membranes as LC3-II, even on maturing autophagosomes. The ratios of these two proteins at different stages of maturation demonstrate that ATG9 proteins are not continuously integrated, but rather are present on the seed vesicles only and become diluted in the rapidly expanding autophagosome membrane. Thus, ATG9 vesicles are the seed membrane from which mammalian autophagosomes form.

cell biology↗

Longitudinal neural and vascular recovery following ultraflexible neural electrode implantation in aged mice

Flexible neural electrodes improve the recording longevity and quality of individual neurons by promoting tissue-electrode integration. However, the intracortical implantation of flexible electrodes inevitably induces tissue damage. Understanding the longitudinal neural and vascular recovery following the intracortical implantation is critical for the ever-growing applications of flexible electrodes in both healthy and disordered brains. Aged animals are of particular interest because they play a key role in modeling neurological disorders, but their tissue-electrode interface remains mostly unstudied. Here we integrate in-vivo two-photon imaging and electrophysiological recording to determine the time-dependent neural and vascular dynamics after the implantation of ultraflexible neural electrodes in aged mice. We find heightened angiogenesis and vascular remodeling in the first two weeks after implantation, which coincides with the rapid increase in local field potentials and unit activities detected by electrophysiological recordings. Vascular remodeling in shallow cortical layers preceded that in deeper layers, which often lasted longer than the recovery of neural signals. By six weeks post-implantation vascular abnormalities had subsided, resulting in normal vasculature and microcirculation. Putative cell classification based on firing pattern and waveform shows similar recovery time courses in fast-spiking interneurons and pyramidal neurons. These results elucidate how structural damages and remodeling near implants affecting recording efficacy, and support the application of ultraflexible electrodes in aged animals at minimal perturbations to endogenous neurophysiology.

neuroscience↗

A high potent synthetic nanobody with broad-spectrum activity neutralizes SARS-Cov-2 virus and Omicron variant through a unique binding mode

The major challenge to control COVID pandemic is the rapid mutation rate of the SARS-Cov-2 virus, leading to the escape of the protection of vaccines and most of the neutralizing antibodies to date. Thus, it is essential to develop neutralizing antibodies with broad-spectrum activity targeting multiple SARS-Cov-2 variants. Here, we reported a synthetic nanobody (named C5G2) obtianed by phage display and subsequent antibody engineering. C5G2 has a single digit nanomolar binding affinity to RBD domain and inhibits its binding to ACE2 with an IC50 of 3.7 nM. Pseudovirus assay indicated that the monovalent C5G2 could protect the cells from the infection of SARS-Cov-2 wild type virus and most of the virus of concern, i.e. Alpha, Beta, Gamma and Omicron variants. Strikingly, C5G2 has the highest potency against Omicron among all the variants with the IC50 of 4.9ng/mL. The Cryo-EM structure of C5G2 in complex with the Spike trimer showed that C5G2 bind to RBD mainly through its CDR3 at a conserved region that not overlapping with the ACE2 binding surface. Additionally, C5G2 bind simultaneously to the neighboring NTD domain of spike trimer through the same CDR3 loop, which may further increase its potency against the virus infection. Third, the steric hindrance caused by FR2 of C5G2 could inhibit the binding of ACE2 to RBD as well. Thus, this triple-function nanobody may be served as an effective drug for the prophylaxis and therapy against Omicron as well as future variants.

biochemistry↗

Visualizing anesthesia-induced vasodilation of cerebral vasculature using multi-exposure speckle imaging

BackgroundAnesthetized animal models are used extensively during neurophysiological and behavioral studies despite systemic effects from anesthesia that undermine both accurate interpretation and translation to awake human physiology. The majority of work examining the impact of anesthesia on cerebral blood flow (CBF) has been restricted to before and after measurements with limited spatial resolution. New MethodWe used multi-exposure speckle imaging (MESI), an advanced form of laser speckle contrast imaging (LSCI), to characterize the dynamics of isoflurane anesthesia induction on cerebral vasculature and blood flow in the mouse brain. ResultsThe large anatomical changes caused by isoflurane are depicted with wide-field imagery and video highlighting the induction of general anesthesia. Within minutes of exposure, both vessel diameter and blood flow increased drastically compared to the awake state and remained elevated for the duration of imaging. An examination of the dynamics of anesthesia induction reveals that blood flow increased faster in arteries than in veins or parenchyma regions. Comparison with Existing MethodsMESI offers robust hemodynamic measurements across large fields-of-view and high temporal resolutions sufficient for continuous visualization of cerebrovascular events featuring major changes in blood flow. ConclusionThe large alterations caused by isoflurane anesthesia to the cortical vasculature and CBF are readily characterized using MESI. These changes are unrepresentative of normal physiology and provide further evidence that neuroscience experiments would benefit from transitioning to un-anesthetized awake animal models.

neuroscience↗

Multimodal mapping of neural activity and cerebral blood flow reveals long-lasting neurovascular dissociations after small-scale strokes

Neurovascular coupling, the close spatial and temporal relationship between neural activity and hemodynamics, is disrupted in pathological brain states. To understand the altered neurovascular relationship in brain disorders, longitudinal, simultaneous mapping of neural activity and hemodynamics is critical yet challenging to achieve. Here, we employ a multimodal neural platform in a mouse model of stroke and realize long-term, spatially-resolved tracking of intracortical neural activity and cerebral blood flow in the same brain regions. We observe a pronounced neurovascular dissociation that occurs immediately after small-scale strokes, becomes the most severe a few days after, lasts into chronic periods, and varies with the level of ischemia. Neuronal deficits extend spatiotemporally whereas restoration of cerebral blood flow occurs sooner and reaches a higher relative value. Our findings reveal the neurovascular impact of mini-strokes and inform the limitation of neuroimaging techniques that infer neural activity from hemodynamic responses.

neuroscience↗