Biology subjects

Lai, H. M.

Publications and source records attributed to Lai, H. M..

7 recordsLinked to original sources

bioRxiv2026

COLOR-3D: a versatile tool for revealing novel 3D histological features

Hematoxylin and eosin (H&E) has been the fundamental method for visualising tissue morphology. Recent advances in tissue clearing and microscopy have enabled the observation of tissue morphology in 3D, but incomplete penetration of nucleic acid dyes has remained the bottleneck. To address this, we develop a new staining chemistry called Cyclodextrin and Organic solvent-assisted deep Labelling of ORgans in 3D (COLOR-3D), which attains the best penetration depth and homogeneity among state-of-the-art methods. We also demonstrate the scalability of COLOR-3D and its compatibility with other staining modalities. To bridge the gap between 3D histology and its wider application in biomedical research and histopathology, we develop a computational pipeline to convert 3D fluorescence images into bright-field H&E images, enabling the creation of a 3D atlas of both normal tissues and pathological specimens. Apart from qualitative observation of tissue morphology, COLOR-3D also enables quantitative analysis for studying biological phenomena. In the mouse liver, we discover rare populations of tetranuclear hepatocytes as well as m16n, t4n and t8n hepatocytes. We also propose the first structural model of the liver lobule based on 3D histology. With a more complete penetration, we reveal the following aging-related changes in tissue microarchitecture, including an increase in extreme nuclear polyploidy, and the disruption of vasculature and portal triad.

pathology↗

bioRxiv2026

nELAVL phosphorylation by CDKL5 regulates inter-condensates composition and communication to promote experience-dependent maturation of the visual cortex

Mutations in Cyclin-Dependent Kinase-Like 5 (CDKL5) cause CDKL5 deficiency disorder (CDD), an X-linked neurodevelopmental condition. Through a phosphoproteomic screen, we identified the neuron-specific nELAVL family of RNA-binding proteins as direct activity-dependent substrates of CDKL5. In support of this regulatory axis, single-nuclei transcriptomics of Cdkl5 knockout (KO) cortices revealed an enriched reduction in activity-dependent mRNAs. Mechanistically, we show that nELAVL proteins undergo phase separation to form biomolecular condensates, the size of which is gated by CDKL5 phosphorylation. Loss of CDKL5 leads to enlarged nELAVL condensates, which exhibit reduced binding affinity for the target mRNA Fos, resulting in its accelerated degradation. This disruption extends to inter-condensate communication: phosphodeficient nELAVL show diminished interaction with P-bodies, which themselves become enlarged in CDD mutant iNeurons. Functionally, the absence of nELAVL phosphorylation recapitulates the deficits in experience-dependent visual function observed in Cdkl5 KO mice. Our findings establish a critical molecular mechanism by which CDKL5-mediated phosphorylation governs mRNA metabolism by tuning the properties of nELAVL condensates and their communication with other biomolecular condensates, ultimately promoting experience-dependent maturation of the visual cortex.

neuroscience↗

bioRxiv2026

Universal nucleic acid preservation in biological fluids with boron clusters

Nucleic acid preservation remains a critical bottleneck for diagnostics and therapeutics, with small molecule inhibitors such as EDTA showing limited spectrum against diverse nucleases, while protein-based alternatives requiring costly cold-chain storage. Here, we report dodecaborate cluster [B12H12]2- as a novel class of pan-nuclease inhibitor with a fundamentally different mechanism--preventing protein-nucleic acid interactions rather than targeting cofactor dependencies. [B12H12]2- inhibited all six DNases and ten RNases tested, making it the broadest spectrum nuclease inhibitor known. Remarkably, its inhibition is reversible via {gamma}-cyclodextrin complexation. We demonstrate that [B12H12]2- preserved a physiological range of DNA for 14 days and that of RNA for 3-7 days at room temperature in human plasma and urine, achieving up to 323,972-fold better RNA retention than controls. A newly developed blood collection tube using [B12H12]2- enables whole-blood circulating cell-free RNA sequencing after 7 days of room-temperature storage, with preserved transcript integrity. Mechanistic studies suggest [B12H12]2- binds to nuclease active sites and creates electrostatic barriers that prevent substrate binding. This chemically stable, indefinitely shelf-stable inhibitor enables cold-chain-free biological sample transport, potentially transforming accessibility of nucleic acid-based diagnostics worldwide. Graphical abstract

bioengineering↗

bioRxiv2025

Psilocybin triggers an activity-dependent rewiring of large-scale cortical networks

Psilocybin holds promise as a treatment for mental illnesses. One dose of psilocybin induces structural remodeling of dendritic spines in the medial frontal cortex in mice. The dendritic spines would be innervated by presynaptic neurons, but the sources of these inputs have not been identified. Here, using monosynaptic rabies tracing, we map the brain-wide distribution of inputs to frontal cortical pyramidal neurons. We discover that psilocybins effect on connectivity is network-specific: strengthening the routing of inputs from perceptual and medial regions (homolog of default mode network) to subcortical targets, while weakening inputs that are part of cortico-cortical recurrent loops. The pattern of synaptic reorganization depends on the drug-evoked spiking activity, because silencing a presynaptic region during psilocybin administration disrupts the rewiring. Collectively, the results reveal the impact of psilocybin on the connectivity of large-scale cortical networks and demonstrate neural activity modulation as an approach to sculpt the psychedelic-evoked neural plasticity.

neuroscience↗

bioRxiv2025

Cerebellum-Targeted Transcranial Focused Ultrasound Stimulation Modulates Hippocampus Neural Activities

Low-intensity transcranial focused ultrasound stimulation (tFUS) has recently emerged as a promising neuromodulation technique due to its non-invasive nature, ability to penetrate deeply, and high spatial resolution. On the other hand, the cerebellum has attracted new interest as a target for neuromodulation. In this study, we investigated how cerebellum-targeted tFUS could modulate hippocampal neural activity. We found that tFUS at pulse repetition frequency (PRF) as high as 10kHz can effectively activate the cerebellum in vivo. Furthermore, PRF at 10kHz, rather than 1kHz, favored remote inhibition of hippocampus neurons. We found that the neuromodulation at hippocampus was mainly mediated by cerebellum cortex than deep cerebellar nucleus. Furthermore, by cFos expression mapping and phase-amplitude coupling analysis, we showed that the hippocampal response at different PRFs resulted from altered neurodynamic interaction rather than overall activation of engaged brain regions. Our results reveal new potential for cerebellum as a neuromodulation target for hippocampus-related functions.

bioengineering↗

bioRxiv2024

Topologically distinct and equivalent multiple efferent arterioles in the human glomeruli

We revisit the assumption that the human glomerulus consists of a capillary network with one afferent arteriole and only one efferent arteriole. Using three-dimensional tissue imaging, we unambiguously demonstrate the existence of multiple efferent arterioles via total reconstruction of a full human glomerular capillary network. Through graph network analysis, we found these multiple efferent arterioles can be topologically distinct or equivalent with reference to the global glomerular capillary network, and were strategically positioned with respect to the afferent arteriole. We also found the extraglomerular connections between two efferent arterioles may have significant contributions to the redistribution of intraglomerular blood flow. These new findings can provide new paradigms in the regulation and dysfunction of glomerular ultrafiltration, the crosstalk between intra- and extraglomerular factors, and the cross-coupling between nephrons.

pathology↗

bioRxiv2024

INSIHGT: Accessible multimodal systems biology with quantitative molecular phenotyping in 3D

Biological systems are complex, encompassing intertwined spatial, molecular and functional features. However, methodological constraints always limit the completeness of information that can be extracted. Here, we report the development of INSIHGT, a minimally perturbative, accessible and cost-efficient three-dimensional (3D) spatial biology method utilizing superchaotropes and host-guest chemistry. This allows highly multiplexed and multi-modal readout of tissue biomolecules in biological systems up to centimeter scales, permitting radio-histological correlation of phosphorylated alpha-synuclein pathologies in human hemi-brainstem. The homogeneous penetration permits reliable semi-quantitative signals in 3D compared to reference signals. Diverse antigens, mRNA transcripts, neurotransmitters, and post-translational and epigenetic modifications, are well-preserved and visualized. INSIHGT also allows multi-round molecular probing for high-dimensional spatial biology and compatibility with downstream traditional histology. With INSIHGT, we mapped previously undescribed podocyte-to-parietal epithelial cell microfilaments and demonstrated their geodesic clustering in mouse glomeruli, and catalogued sparsely located neurofilament-intensive inclusion bodies in the human cerebellum, and identified NPY-proximal cell types defined by spatial morpho-proteomics in mouse hypothalamus. We anticipate INSIHGT can form the foundations for 3D spatial multi-omics technology development and holistic systems biology studies.

bioengineering↗