Revealing and mitigating the inhibitory effect of serotonin on HRP-mediated protein labelling DOI Creative Commons
Zora Chui-Kuen Chan, Qi Cheng, Yanbin Cai

и другие.

Scientific Reports, Год журнала: 2024, Номер 14(1)

Опубликована: Дек. 30, 2024

Proximity-dependent biotinylation coupled with mass spectrometry enables the characterization of subcellular proteomes. This technique has significantly advanced neuroscience by revealing sub-synaptic protein networks, such as synaptic cleft and post-synaptic density. Profiling proteins at this detailed level is essential for understanding molecular mechanisms neuronal connectivity transmission. Despite its recent successful application to various types, proximity labelling yet be employed study serotonin system. In study, we uncovered an unreported inhibitory mechanism on horseradish peroxidase (HRP)-based biotinylation. Our result showed that reduces levels across Biotin-XX-tyramide (BxxP) concentrations in HEK293T cells primary neurons, whereas dopamine exerts minimal interference, highlighting specificity inhibition. To counteract inhibition, demonstrated Dz-PEG, aryl diazonium compound consumes through azo-coupling reaction, restores efficiency. Label-free quantitative proteomics confirmed inhibits biotinylation, Dz-PEG effectively reverses These findings highlight importance accounting neurotransmitter interference proximity-dependent studies, especially cell-type specific profiling neuroscience. Additionally, provided a potential strategy mitigate these challenges, thereby enhancing accuracy reliability studies.

Язык: Английский

Progress toward a comprehensive brain protein interactome DOI

Vy Dang,

Brittney Voigt, Edward M. Marcotte

и другие.

Biochemical Society Transactions, Год журнала: 2025, Номер 53(01)

Опубликована: Фев. 12, 2025

Protein–protein interactions (PPIs) in the brain play critical roles across all aspects of central nervous system, from synaptic transmission, glial development, myelination, to cell-to-cell communication, and more. Understanding these is crucial for deciphering neurological mechanisms underlying biochemical machinery affected disorders. Recently, advances proteomics techniques have significantly enhanced our ability study among proteins expressed brain. Here, we review some high-throughput studies characterizing PPIs, using affinity purification, proximity labeling, co-fractionation, chemical cross-linking mass spectrometry methods, as well yeast two-hybrid assays. We present current state field, discuss challenges, highlight promising future directions.

Язык: Английский

Процитировано

0
Three systems of circuit formation: assembly, updating and tuning DOI
Dániel L. Barabási, André Ferreira Castro, Florian Engert

и другие.

Nature reviews. Neuroscience, Год журнала: 2025, Номер unknown

Опубликована: Фев. 24, 2025

Язык: Английский

Процитировано

0
Rewiring an olfactory circuit by altering the combinatorial code of cell-surface proteins DOI Creative Commons
Cheng Lyu, Zhuoran Li,

Chuanyun Xu

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2025, Номер unknown

Опубликована: Март 2, 2025

Abstract Proper brain function requires the precise assembly of neural circuits during development. Despite identification many cell-surface proteins (CSPs) that help guide axons to their targets 1,2 , it remains largely unknown how multiple CSPs work together assemble a functional circuit. Here, we used synaptic partner matching in Drosophila olfactory circuit 3,4 address this question. By systematically altering combination differentially expressed single receptor neuron (ORN) type, which senses male pheromone inhibits male-male courtship, switched its connection from endogenous postsynaptic projection (PN) type nearly completely new PN promotes courtship. To achieve switch, deduced combinatorial code including mediate both attractive and repulsive interactions between partners 5,6 . The anatomical switch changed odor response markedly increased We generalized three manipulation strategies rewiring successfully rewire second ORN distinct types. This demonstrates manipulating small set is sufficient respecify connections, paving ways explore systems evolve through changes connectivity.

Язык: Английский

Процитировано

0
Protocol for cell-type-specific single-cell labeling and manipulation in Drosophila using a sparse driver system DOI Creative Commons

Chuanyun Xu,

Zhuoran Li, Liqun Luo

и другие.

STAR Protocols, Год журнала: 2025, Номер 6(1), С. 103694 - 103694

Опубликована: Март 1, 2025

Here, we present a protocol for cell-type-specific single-cell labeling and manipulation in Drosophila using sparse driver system. We describe steps generating constructs fly lines, titrating heat-shocked durations precise temporal control desired sparsity, co-expressing multiple transgenes experiments. demonstrate that this generalizable toolkit enables tunable multi-color staining, trans-synaptic tracing, manipulation, cell-autonomous gene function analysis. For complete details on the use execution of protocol, please refer to Xu et al.1.

Язык: Английский

Процитировано

0
Dimensionality reduction simplifies synaptic partner matching in an olfactory circuit DOI Creative Commons
Cheng Lyu, Zhuoran Li,

Chuanyun Xu

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Авг. 27, 2024

Abstract The distribution of postsynaptic partners in three-dimensional (3D) space presents complex choices for a navigating axon. Here, we discovered dimensionality reduction principle establishing the 3D glomerular map fly antennal lobe. Olfactory receptor neuron (ORN) axons first contact partner projection (PN) dendrites at 2D spherical surface lobe during development, regardless whether adult glomeruli are or interior Along surface, each ORN type take specific 1D arc-shaped trajectory that precisely intersects with their PN dendrites. Altering axon trajectories compromises synaptic matching. Thus, search problem is reduced to 1D, which simplifies matching and may generalize wiring process more brains.

Язык: Английский

Процитировано

1
The sparse driver system for in vivo single-cell labeling and manipulation in Drosophila DOI Creative Commons

Chuanyun Xu,

Liqun Luo

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Дек. 4, 2024

In this protocol, we introduce a sparse driver system for cell-type specific single-cell labeling and manipulation in

Язык: Английский

Процитировано

0
Serotonin-Induced Inhibition of HRP-Mediated Proximity Labelling DOI Creative Commons
Zora Chui-Kuen Chan, Qi Cheng, Yanbin Cai

и другие.

Research Square (Research Square), Год журнала: 2024, Номер unknown

Опубликована: Дек. 16, 2024

Abstract Proximity-dependent biotinylation coupled with mass spectrometry enables the characterization of subcellular proteomes. This technique has significantly advanced neuroscience by revealing sub-synaptic protein networks, such as synaptic cleft and post-synaptic density. Profiling proteins at this detailed level is essential for understanding molecular mechanisms neuronal connectivity transmission. Despite its recent successful application to various types, proximity labelling yet be employed study serotonin system. In study, we uncovered an unreported inhibitory mechanism on horseradish peroxidase (HRP)-based biotinylation. Our result showed that reduces levels across Biotin-XX-tyramide (BxxP) concentrations in HEK293T cells primary neurons, whereas dopamine exerts minimal interference, highlighting specificity inhibition. To counteract inhibition, demonstrated Dz-PEG, aryl diazonium compound consumes through azo-coupling reaction, restores efficiency. Label-free quantitative proteomics confirmed inhibits biotinylation, Dz-PEG effectively reverses These findings highlight importance accounting neurotransmitter interference proximity-dependent studies, especially cell-type specific profiling neuroscience. Additionally, provided a potential strategy mitigate these challenges, thereby enhancing accuracy reliability studies.

Язык: Английский

Процитировано

0
Revealing and mitigating the inhibitory effect of serotonin on HRP-mediated protein labelling DOI Creative Commons
Zora Chui-Kuen Chan, Qi Cheng, Yanbin Cai

и другие.

Scientific Reports, Год журнала: 2024, Номер 14(1)

Опубликована: Дек. 30, 2024

Proximity-dependent biotinylation coupled with mass spectrometry enables the characterization of subcellular proteomes. This technique has significantly advanced neuroscience by revealing sub-synaptic protein networks, such as synaptic cleft and post-synaptic density. Profiling proteins at this detailed level is essential for understanding molecular mechanisms neuronal connectivity transmission. Despite its recent successful application to various types, proximity labelling yet be employed study serotonin system. In study, we uncovered an unreported inhibitory mechanism on horseradish peroxidase (HRP)-based biotinylation. Our result showed that reduces levels across Biotin-XX-tyramide (BxxP) concentrations in HEK293T cells primary neurons, whereas dopamine exerts minimal interference, highlighting specificity inhibition. To counteract inhibition, demonstrated Dz-PEG, aryl diazonium compound consumes through azo-coupling reaction, restores efficiency. Label-free quantitative proteomics confirmed inhibits biotinylation, Dz-PEG effectively reverses These findings highlight importance accounting neurotransmitter interference proximity-dependent studies, especially cell-type specific profiling neuroscience. Additionally, provided a potential strategy mitigate these challenges, thereby enhancing accuracy reliability studies.

Язык: Английский

Процитировано

0