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Chu, L.-A.

Publications and source records attributed to Chu, L.-A..

12 recordsLinked to original sources

Long-term memory performance optimization via Neural network-based curve fitting in Drosophila

Long-term memory (LTM) formation typically requires extensive training or highly salient experiences, limiting learning efficiency. Operant conditioning is generally thought to produce stronger memory than classical conditioning because of its active learning component. Unexpectedly, however, laser-based social conditioning in Drosophila melanogaster revealed that while classical paradigms yielded lower short-term memory (STM) scores but higher LTM retention, operant paradigms exhibited higher STM scores followed by rapid LTM decay. To resolve this discrepancy, we employed the AI Complex Systems Response (AI-CSR) framework, which reconstructs high-dimensional learning landscapes from sparse sampling and predicts globally optimal training conditions. Following AI-CSR optimization, operant conditioning produced a twofold increase in LTM scores, yielding the strongest 24-hour social memory performance reported in flies to date and revealing the expected superiority of active learning, which had conversely shown poorer performance under standard training protocols. In contrast, AI-CSR did not further enhance classical conditioning performance but reduced training time by 50%. Single-cell RNA sequencing revealed expanded neuronal recruitment marked by the activation and inhibition of various gene combinations. Together, these findings link circuit-level reorganization with molecular programs underlying efficient long-term memory and demonstrate how AI-guided optimization can uncover latent learning capacity in biological systems.

animal behavior and cognition↗

High-resolution volumetric intravital imaging reveals asymmetric serotonin-dependent post-shock activity in the Drosophila brain

Volumetric intravital imaging of adult Drosophila brains remains challenging due to optical scattering and speed limitations of conventional microscopy. Here, we present V-shape Bessel-beam light-sheet microscopy (vSPIM), an upright, high-speed platform achieving subcellular resolution across large, opaque volumes in vivo. Through real-time calcium imaging in adult Drosophila, vSPIM uncovered two unprecedented phenomena: first, a complex ensemble of approximately 150 olfactory projection neuron boutons (more than 400 per hemisphere) resolved 18 distinct odor valence coding patterns within the dense mushroom body calyx, aligning with connectomic architecture. Second, vSPIM revealed a previously uncharacterized asymmetric, serotonin-dependent post-shock firing (PsF) response. This PsF activity emerged within the mushroom body and dopaminergic neurons only after repetitive aversive stimulation, distinct from canonical dopamine-driven reinforcement signals. Finally, we demonstrated the versatility of vSPIM by tracking corneal endothelial dynamics during mouse wound healing. Overall, vSPIM establishes a scalable framework for intravital imaging, bridging the gap between optical innovation and circuit physiology in adult tissues.

neuroscience↗

Robust volumetric multiplex staining of centimeter-scale FFPE tissues guided by neural network-based optimization

Archived formalin-fixed, paraffin-embedded (FFPE) tissues, including brain-bank specimens, are valuable resources for disease research, but rapid cyclic immunolabeling of intact specimens remains challenging. Here, we present FIDELITY, an epoxy-free workflow that balances optical transparency, immunostaining performance, and tissue integrity for multi-cycle protein profiling in centimeter-thick FFPE specimens. Neural network-guided optimization identified an SDS-glycine formulation that integrates delipidation and epitope retrieval while preserving tissue stiffness, replacing prolonged epoxy reinforcement and subsequent high-temperature antigen retrieval with a single aqueous pretreatment. Application to human Alzheimer's disease specimens enabled volumetric imaging and quantitative analysis while retaining compatibility with conventional histopathology. Its compatibility with active electrophoretic staining further supported rapid labeling and five rounds of immunolabeling in a whole FFPE mouse brain. Two-round labeling of a human glioma specimen further demonstrated the applicability of the cyclic workflow to human pathological tissue. Together, FIDELITY provides a practical platform for large-volume, multiplexed protein profiling of intact FFPE specimens.

neuroscience↗

Latent-centric Isotropic Resolution Enhancement for Expansion Microscopy Imaging via Neural Compression and Self-supervised Learning

Expansion microscopy (ExM) enables nanoscale imaging for disease characterization. However, whole-organ analyses remain limited by several challenges. Current super-resolution methods either require high-resolution ground-truth data or assume spatially uniform point spread functions--assumptions that rarely hold in whole-organ imaging with depth-varying aberrations and illumination drift. Existing methods also worsen storage demands by inflating already multi-terabyte datasets without using neural compression. We propose a single-stage, self-supervised framework that addresses both resolution anisotropy and storage constraints through compression-aware isotropic super-resolution. Our approach combines a 2D lateral encoder that operates directly on raw slices to avoid memory limits with a lightweight volumetric decoder that preserves cross-slice continuity. A vector-quantized variational autoencoder (VQ-VAE) provides an information-sufficient bottleneck, achieving up to 128x slice compression and up to 8x axial resolution enhancement. This latent-centric design yields approximately 1000x reduction in storage compared with storing fully isotropic volumes. The framework achieves higher GPU throughput, lower memory usage, and stronger multi-GPU scalability than prior methods. By designating compressed latent space as the native storage format, it enables efficient on-demand isotropic reconstruction directly from compact representations. This combination of isotropic enhancement and neural compression framework therefore makes large-scale, whole-organ ExM analysis practical while maintaining analysis-ready accessibility, addressing a bottleneck in translating ExM to clinical biomarker discovery.

bioinformatics↗

Dopaminergic Modulation of Mushroom Body Output Neurons Mediates Nociception-Induced Escape in Drosophila

In Drosophila, noxious heat is detected by peripheral nociceptors expressing transient receptor potential (TRP) channels, including Painless and TrpA1, and rapidly triggers escape behavior. Although peripheral transduction has been defined in detail, the central circuits and neuromodulatory mechanisms that translate nociceptor activity into escape decisions remain poorly understood. Here, we combine targeted behavioral perturbations with anatomical tracing to delineate a nociception-to-escape pathway that engages dopaminergic modulation of mushroom body (MB) output. Kir2.1-mediated silencing across candidate neurotransmitter systems revealed a specific requirement for MB-innervating dopaminergic neurons (DANs)--particularly subsets within the protocerebral posterior lateral 1 (PPL1) and protocerebral anterior medial (PAM) clusters--for robust nociception-induced escape. Anterograde trans-Tango tracing from painless- and trpA1-expressing nociceptors labeled these MB dopaminergic neurons as direct postsynaptic partners, consistent with convergence of distinct nociceptor inputs onto a shared dopaminergic pathway. Finally, silencing a subset of mushroom body output neurons (MBONs) delayed escape without overtly disrupting baseline locomotion, supporting a model in which dopaminergic signaling recruits MB output to shape defensive action selection. Together, our results define a multi-layer circuit motif linking peripheral nociception to MB-dependent escape and provide a framework for dissecting how neuromodulation gates rapid defensive behaviors.

neuroscience↗

Expansion microscopy for super-resolution imaging of collagen-abundant tissues

Expansion microscopy (ExM) is popular for three-dimensional ultrastructural imaging of cultured cells and tissue slices at nanoscale resolution with conventional microscopes via physical expansion of biological tissues. However, the application of this technology to collagen-abundant thick tissues is challenging. We demonstrate a new method, collagen expansion microscopy (ColExM), optimized for expanding tissues containing more than 70% collagen. ColExM succeeded in 4.5-fold linear expansion with minimal structural distortion of corneal and skin tissues. It was also compatible with immunostaining, allowing super-resolution visualization of three-dimensional neural structures innervating hair follicles and corneas. With ColExM, we succeeded in identifying individual mitochondria and previously unrecognized dendritic spine-like structures of corneal nerves. ColExM also enabled fine mapping of structural rearrangement of tight junctions and actin cytoskeletons. Therefore, this method can facilitate the exploration of three-dimensional nanoscale structures in collagen-rich tissues.

bioengineering↗

Video-rate three-photon imaging in deep Drosophila brain based on a single Cr:forsterite oscillator

We have demonstrated 30-Hz three-photon imaging using a single 24-MHz mode-locked Cr:forsterite oscillator with a center wavelength at 1260 nm. By managing the dispersion distribution in the resonator using double-chirped mirrors, we have produced 32-fs pulses with 22-nJ pulse energy. Using the oscillator as a driving source, we have realized multi-color three-photon images using a GFP-labeled Drosophila brain and an AF647-labeled mouse brain. To demonstrate the capability of deep-tissue imaging, we have obtained a 10-times higher SBR from the three-photon images than the two-photon results at different depths in a GFP-labeled Drosophila brain dissection. Furthermore, we have shown the impact of excitation pulse width on three-photon deep-tissue imaging. Our results indicate the superiority of using shorter pulses for deeper-tissue imaging, especially in the Drosophila brain. In addition, we have recorded the three-photon calcium imaging in vivo from the Drosophila mushroom body in response to external electric shocks. We believe our demonstration provides a robust approach for high-speed three-photon microscopy applications, especially for intravital investigations in the Drosophila brain.

bioengineering↗

A Weakly Supervised U-Net Model for Precise Whole Brain Immunolabeled Cell Detection

Cell segmentations low precision due to the intensity differences hinders widespread use of whole brain microscopy imaging. Previous studies used ResNet or CNN to account for this problem, but are unapplicable to immunolabeled signals across samples. Here we present a semiauto ground truth generation and weakly-supervised U-Net-based Deep-learning precise segmentation pipeline for whole brain immunopositive c-FOS signals, which reveals the distinct neural activity maps with different social motivations.

neuroscience↗

A versatile method to archive and delipidate brain tissues for ensuing multiplexed immunolabeling and organ-level imaging

Tissue clearing and labeling techniques have revolutionized brain-wide imaging and analysis, yet their application to clinical formalin-fixed paraffin-embedded (FFPE) blocks remains challenging. We introduce HIF-Clear, a novel method for efficiently clearing and labeling centimeter-thick FFPE specimens using elevated temperature and concentrated detergents. HIF-Clear with multi-round immunolabeling reveals neuron circuitry regulating multiple neurotransmitter systems in a whole FFPE mouse brain, and is able to be used as the evaluation of disease treatment efficiency. HIF-Clear also supports expansion microscopy and can be performed on a non-sectioned 15-year-old FFPE specimen, as well as a 3-month formalin-fixed mouse brain. Thus, HIF-Clear represents a feasible approach for researching archived FFPE specimens for future neuroscientific and 3D neuropathological analyses.

bioengineering↗

CyDAP-A fluorescent probe for cytosolic dopamine detection

Dopamine (DA) is an essential neurotransmitter modulating motor and cognitive functions. Several neurological disorders, including Parkinsons disease (PD) and drug addiction, are the result of DA system dysfunction; however, it remains incomplete understood of why DA neuron is selectively more vulnerable than other neurons. Here we utilize the spectral feature of human MAO B (monoamine oxidase B) to design a genetic-amenable, GFP-based fluorescent probe CyDAP. Upon genetic and pharmacological manipulations to elevate the cytosolic DA levels in cells and Drosophila models, CyDAP shows enhanced GFP emission, suggesting this probe is feasible for DA detection. Furthermore, we observe that expressing human -Synuclein in Drosophila elicited GFP emission from CyDAP, suggesting a link between cytosolic DA imbalance and regional vulnerability in PD context. Importantly, CyDAP can detect the change of cytosolic DA in live Drosophila brains, as demonstrated by time-lapse and the 4D light-sheet confocal recording. CyDAP may serve as a tool for evaluating metabolic deregulation of DA in brain models of PD and other DA system-related psychiatric disorders.

neuroscience↗

Visualization of axonal protein allocation in Drosophila with whole brain localization microscopy

Long-term memory (LTM) formation requires learning-induced protein synthesis in specific neurons and synapses within a neural circuit. Precisely how neural activity allocates new proteins to specific synaptic ensembles, however, remains unknown. We developed a deep-tissue super-resolution imaging tool suitable for single-molecule localization in intact adult Drosophila brain, and focused on the axonal protein allocation in mushroom body (MB), a central neuronal structure involved in olfactory memory formation. We found that insufficient training suppresses LTM formation by inducing the synthesis of vesicular monoamine transporter (VMAT) proteins within a dorsal paired medial (DPM) neuron, which innervates all axonal lobes of the MB. Surprisingly, using our localization microscopy, we found that these learning-induced proteins are distributed only in a subset of DPM axons in specific sectors along the MB lobes. This neural architecture suggests that sector-specific modulation of neural activity from MB neurons gates consolidation of early transient memory into LTM.

neuroscience↗

Asymmetric ephaptic inhibition between compartmentalized olfactory receptor neurons

In the Drosophila antenna, different subtypes of olfactory receptor neurons (ORNs) housed in the same sensory hair (sensillum) can inhibit each other non-synaptically. However, the mechanisms underlying this unusual form of lateral inhibition remain unclear. Here we use recordings from pairs of sensilla impaled by the same tungsten electrode to prove that direct electrical (\"ephaptic\") interactions mediate lateral inhibition between ORNs. Intriguingly, within individual sensilla, we find that ephaptic lateral inhibition is asymmetric such that one ORN exerts greater influence onto its neighbor. Serial block-face scanning electron microscopy of genetically identified ORNs and circuit modeling indicate that asymmetric lateral inhibition reflects a surprisingly simple mechanism: the physically larger ORN in a pair corresponds to the dominant neuron in ephaptic interactions. Thus, morphometric differences between compartmentalized ORNs account for highly specialized inhibitory interactions that govern information processing at the earliest stages of olfactory coding.

neuroscience↗