Biology subjects

Ho, Y.-P.

Publications and source records attributed to Ho, Y.-P..

6 recordsLinked to original sources

bioRxiv2026

RbohF-mediated ROS production is required for tapetal PCD and pollen maturation in rice

Reactive oxygen species (ROS) function as critical signaling molecules during plant reproductive development, particularly in regulating tapetal programmed cell death (PCD). However, the molecular mechanisms controlling ROS production in rice anthers remain incompletely understood. In this study, we characterized the role of RbohF, a rice respiratory burst oxidase homolog, using CRISPR/Cas9 knockout lines. Loss of RbohF resulted in severe defects in anther development, including impaired pollen maturation and reduced fertility. Histological analyses revealed progressive degeneration of microspores and abnormal tapetal persistence in the mutant. ROS staining demonstrated that superoxide and hydrogen peroxide accumulation was significantly reduced and delayed in rbohf anthers, particularly at the young microspore stage. Consistently, TUNEL assays showed delayed tapetal PCD in the mutant, indicating that RbohF-mediated ROS production is required for the timely initiation of tapetal degeneration. Gene expression analyses further suggested that RbohF functions downstream of major transcriptional regulators of pollen development. Notably, the male-sterile phenotype of rbohf mutants was exacerbated at cooler temperatures, highlighting a role for RbohF in buffering environmental stress. Together, these findings establish RbohF as a key regulator of ROS homeostasis, integrating developmental and environmental signals to ensure proper pollen development in rice.

developmental biology↗

bioRxiv2025

MusTer: A Generalizable Microfluidic Platform Combining Multi-Parametric Droplet Sorting and Multi-Droplet Merging in Single-Cell Sequencing

Droplet microfluidics is a core technology that powers high-throughput single cell sequencing. However, the current generation of single-cell microfluidics faces notable limitations, including cell aggregation, suboptimal on-chip reactions that compromise experimental outcomes and elevate background noise, as well as a dependence on costly commercial barcode beads. To address these challenges, we present MusTer, an integrated next-generation platform with multi-parametric singlet droplet sorting and triple-droplet merging capability. MusTers multi-parametric singlet sorting module enables in-line droplet analysis of intrinsic fluorescence peak amplitude, width and interval from single-nucleus- (singlet) or multiple-nuclei (multiplet)-encapsulating droplets, subsequently allowing an effective separation of the singlet droplets from multiplet droplets and empty droplets. MusTers triple-droplet merging module enables precise multi-step reactions, with each step performed under its own optimal conditions, thereby significantly enhancing experimental flexibility and efficiency. We validated MusTers performance by performing single-cell ATAC-seq on maize leaves. The results demonstrate that MusTer significantly reduces the doublet rate, enhances the signal-to-noise ratio, and yields improved cell clustering compared with traditional methods. These results validate MusTers capability to overcome key limitations in droplet-based single-cell analysis, effectively enhancing data quality and reliability, and also paves the way for its use in other challenging sample types and multi-step single-cell assays.

bioengineering↗

bioRxiv2024

A Small-molecule Antagonist Radiotracer for Positron Emission Tomography Imaging of the Mu Opioid Receptor.

The opioid crisis is a catastrophic health emergency catalyzed by the misuse of opioids that target and activate the mu opioid receptor. Traditional radioligands used to study the mu opioid receptor are often tightly regulated owing to their abuse and respiratory depression potential. In the present study, we sought to design and characterize a library of 24 non-agonist ligands for the mu opioid receptor. Ligands were evaluated for the binding affinity, intrinsic activity, and predicted blood-brain barrier permeability. Several ligands demonstrated single-digit nM binding affinity for the mu opioid receptor while also demonstrating selectivity over the delta and kappa opioid receptors. The antagonist behavior of 1A and 3A at the mu opioid receptor indicate that these ligands would likely not induce opioid-dependent respiratory depression. Therefore, these ligands can enable a safer means to interrogate the endogenous opioid system. Based on binding affinity, selectivity, and potential off-target binding, [11C]1A was prepared via metallophotoredox of the aryl-bromide functional group to [11C]methyl iodide. The nascent radiotracer demonstrated brain uptake in a rhesus macaque model and accumulation in the caudate and putamen. Naloxone was able to reduce [11C]1A binding, though the interactions were not as pronounced as naloxones ability to displace [11C]carfentanil. These results suggest that GSK1521498 and related congeners are amenable to radioligand design and can offer a safer way to query opioid neurobiology.

neuroscience↗

bioRxiv2023

High-throughput generic single-entity sequencing using droplet microfluidics

Single-cell sequencing has revolutionized our understanding of cellular heterogeneity by providing a micro-level perspective over the past decades. Although heterogeneity is essential for various biological communities, the currently demonstrated platform predominantly focuses on eukaryotic cells without cell walls and their transcriptomics 1,2, leaving significant gaps in the study of omics from other single biological entities such as bacteria and viruses. Due to the difficulty of isolating and acquiring their DNA3, contemporary methodologies for the characterization of generic biological entities remain conspicuously constrained, with low throughput4, compromised lysis efficiency5, and highly fragmented genomes6. Herein, we present the Generic Single Entity Sequencing platform (GSE-Seq), which boasts ample versatility, high throughput, and high coverage, and is enabled by an innovative workflow, addressing the critical challenges in single entities sequencing: (1) one-step manufacturing of massive barcodes, (2) degradable hydrogel-based in situ sample processing and whole genome amplification, (3) integrated in-drop library preparation, (4) compatible long-read sequencing. By GSE-Seq, we have achieved a significant milestone by enabling high-throughput, long-read single-entity profiling of dsDNA and ssDNA from single virus sequencing (SV-seq) and single bacteria sequencing (SB-seq) of the human gut and marine sediment for the first time. Notably, our analysis uncovered previously overlooked viral and bacterial dark matter and phage-host interactions. In summary, the presented conceptually new workflow offers a toolbox based on droplet microfluidics to tackle the persistent challenges in high-throughput profiling to generic applications, which hold immense promise for diverse biological entities, especially hard-to-lyse cells.

bioengineering↗

bioRxiv2021

Metabolic Regulation of Mitochondrial Morphologies in Pancreatic Beta Cells: Bioenergetics-Mitochondrial Dynamics Coupling

Cellular bioenergetics and mitochondrial dynamics are crucial for the secretion of insulin by pancreatic beta cells in response to elevated blood glucose concentrations. To obtain better insights into the interactions between energy production and mitochondrial fission/fusion dynamics, we combine live-cell mitochondria imaging with biophysical-based modeling and network analysis to elucidate the principle regulating mitochondrial morphology to match metabolic demand in pancreatic beta cells. A minimalistic differential equation-based model for beta cells was constructed to include glycolysis, oxidative phosphorylation, simple calcium dynamics, and graph-based fission/fusion dynamics controlled by ATP synthase flux and proton leak flux. The model revealed that mitochondrial fission occurs in response to hyperglycemia, starvation, ATP synthase inhibition, uncoupling, and diabetic condition, in which the rate of proton leak exceeds the rate of mitochondrial ATP synthesis. Under these metabolic challenges, the propensities of tip-to-tip fusion events simulated from the microscopic images of the mitochondrial networks were lower than those in the control group and prevented mitochondrial network formation. The modeling and network analysis could serve as the basis for further detailed research on the mechanisms of bioenergetics and mitochondrial dynamics coupling.

systems biology↗

bioRxiv2018

Low-Density Lipoprotein Receptor-Mediated Lipidome-Transcriptome Reprogramming Impulses to Cisplatin Insensitivity

Platinum-based therapy remains the cornerstone for cancer patient management; however, its efficacy varies. Theis study demonstrated the differential expressions of low-density lipoprotein receptor (LDLR) in subtypes of epithelial ovarian carcinoma (EOC) determines cisplatin sensitivity. Its sensitive in serous EOCs (low LDLR), where insensitive in endometrioid and clear cell EOCs (high LDLR). Meanwhile, knocked-down or overexpressed LDLR in EOC could reversed the chemosensitivity pattern both in vitro and in vivo. Mechanistic dissection with transcriptome vs. lipidome trans-omics analyses elucidated the LDLR[->]LPC (Lyso-PhosphotidylCholine)[->]FAM83B (phospholipase-related)[->]FGFRs (cisplatin sensitivity and phospholipase-related) regulatory axis in cisplatin insensitivity. Implementing LPC-liposome encapsulated cisplatin could facilitate DNA-adduct formation via lipid droplets (LDs) delivery. Furthermore, Bioinformatics analyses found that the LDL/R[->]LD homeostasis alteration is critical for therapeutic prognosis. Lastly, using LPC-liposome-cisplatin improved cisplatin sensitivities in gastric cancer, renal cell carcinoma, hepatocellular carcinoma, cholangiocarcinoma, and pancreatic adenocarcinoma cells. In conclusion, this report discovered a LDL/R-reprogrammed transcriptome-lipidome network, by which impulses platinum insensitivity and disease outcome. The drug specific lipidome for liposome manufacture might be an efficienct pharmaceutics strategy for chemoagents.\n\nSignificanceLDLR reprograms cellular lipidome and transcriptome profiles to determines chemotherapy therapeutic efficacy. The LDLR-reduced LPC abundance disturbs phospholipids homeostasis of Lands cycle in LD, by which attenuates intracellular platinum transportation for DNA-adduct formation. Targeting LDLR-LD-lipidome with LPC-liposome-platinum could boost therapeutic efficacy for insensitivity.

cancer biology↗