Monitoring of activity-driven trafficking of endogenous synaptic proteins through proximity labeling DOI Creative Commons
Carlos Pascual-Caro, Jaime de Juan‐Sanz

PLoS Biology, Journal Year: 2024, Volume and Issue: 22(10), P. e3002860 - e3002860

Published: Oct. 28, 2024

To enable transmission of information in the brain, synaptic vesicles fuse to presynaptic membranes, liberating their content and exposing transiently a myriad vesicular transmembrane proteins. However, versatile methods for quantifying translocation endogenous proteins during neuronal activity remain unavailable, as fast dynamics vesicle cycling difficult specific isolation trafficking such transient surface exposure. Here, we developed novel approach using cleft proximity labeling capture quantify activity-driven at synapse. We show that accelerating biotinylation times match exocytosis allows capturing exposed neural activity, enabling first time study nearly every protein. As proof-of-concept, further applied this technology obtain direct evidence noncanonical proteins, ATG9A NPTX1, which had been proposed traffic but proof not yet shown. The technological advancement presented here will facilitate future studies dissecting molecular identity exocytosed synapse helping define machinery sustains neurotransmission mammalian brain.

Language: Английский

The Role of ATG9 Vesicles in Autophagosome Biogenesis DOI Creative Commons
Elisabeth Holzer, Sascha Martens, Susanna Tulli

et al.

Journal of Molecular Biology, Journal Year: 2024, Volume and Issue: 436(15), P. 168489 - 168489

Published: Feb. 10, 2024

Autophagy mediates the degradation and recycling of cellular material in lysosomal system. Dysfunctional autophagy is associated with a plethora diseases including uncontrolled infections, cancer neurodegeneration. In macroautophagy (hereafter autophagy) this encapsulated double membrane vesicles, autophagosomes, which form upon induction autophagy. The precursors to referred as phagophores, first appear small flattened cisternae, gradually enclose cargo they grow. assembly phagophores during initiation has been major subject investigation over past decades. A special focus ATG9, only conserved transmembrane protein among core machinery. majority ATG9 localizes Golgi-derived vesicles. Here we review recent advances breakthroughs regarding our understanding how vesicles it resides serve assemble machinery establish contact sites for autophagosome biogenesis. We also highlight open questions field that need be addressed years come.

Language: Английский

Citations

24
Plekhg5 controls the unconventional secretion of Sod1 by presynaptic secretory autophagy DOI Creative Commons

Amy-Jayne Hutchings,

Bita Hambrecht,

Alexander Veh

et al.

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: Oct. 4, 2024

Language: Английский

Citations

5
Emerging roles of ATG9/ATG9A in autophagy: implications for cell and neurobiology DOI Creative Commons
Ji-Young Choi,

Haeun Jang,

Xuan Zhao

et al.

Autophagy, Journal Year: 2024, Volume and Issue: 20(11), P. 2373 - 2387

Published: Aug. 4, 2024

Atg9, the only transmembrane protein among many autophagy-related proteins, was first identified in year 2000 yeast. Two homologs of ATG9A and ATG9B, have been found mammals. While ATG9B shows a tissue-specific expression pattern, such as placenta pituitary gland, is ubiquitously expressed. Additionally, deficiency leads to severe defects not at molecular cellular levels but also organismal level, suggesting key fundamental roles for ATG9A. The subcellular localization on small vesicles its functional relevance autophagy suggested potential role lipid supply during autophagosome biogenesis. Nevertheless, precise autophagic process has remained long-standing mystery, especially neurons. Recent findings, however, including structural, proteomic, biochemical analyses, provided new insights into function expansion phagophore membrane. In this review, we aim understand various aspects ATG9 (in invertebrates plants)/ATG9A mammals), localization, trafficking, other functions, nonneuronal cells neurons by comparing recent discoveries related ATG9/ATG9A proposing directions future research.

Language: Английский

Citations

4
MINFLUX Nanoscopy: A “Brilliant” Technique Promising Major Breakthrough DOI Creative Commons
Marco Salerno, Virginia Bazzurro, Elena Angeli

et al.

Microscopy Research and Technique, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 21, 2025

ABSTRACT MINFLUX nanoscopy relies on the localization of single fluorophores with expected ~ 2 nm precision in 3D mapping, roughly one order magnitude better than standard stimulated emission depletion microscopy or stochastic optical reconstruction microscopy. This “brilliant” technique takes advantage specialized principles and algorithms that require only dim fluorescence signals a minimum flux photons; hence name follows. With this level performance, imaging tracking should allow for routine study biological processes down to molecular scale, revealing previously unresolved details cell structures, such as organization calcium channels muscle cells clustering receptors synapses. Whereas high is definitely strength technique, limitations challenges also exist, especially labeling procedures aiming at appropriate density on/off switching kinetics. primer presents some significant results achieved so far highlights specific operational crucial technique.

Language: Английский

Citations

0
Neuronal autophagy in the control of synapse function DOI Creative Commons
Anna Karpova, P. Robin Hiesinger, Marijn Kuijpers

et al.

Neuron, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 1, 2025

Neurons are long-lived postmitotic cells that capitalize on autophagy to remove toxic or defective proteins and organelles maintain neurotransmission the integrity of their functional proteome. Mutations in genes cause congenital diseases, sharing prominent brain dysfunctions including epilepsy, intellectual disability, neurodegeneration. Ablation core neurons glia disrupts normal behavior, leading motor deficits, memory impairment, altered sociability, which associated with defects synapse maturation, plasticity, neurotransmitter release. In spite importance for physiology, substrates neuronal mechanisms by affect synaptic function health disease remain controversial. Here, we summarize current state knowledge autophagy, address existing controversies inconsistencies field, provide a roadmap future research role control function.

Language: Английский

Citations

0
The life and times of brain autophagic vesicles DOI
Lisa Gambarotto, Erin Wosnitzka, Vassiliki Nikoletopoulou

et al.

Journal of Molecular Biology, Journal Year: 2025, Volume and Issue: unknown, P. 169105 - 169105

Published: March 1, 2025

Language: Английский

Citations

0
The synaptic vesicle cluster as a controller of pre‐ and postsynaptic structure and function DOI Creative Commons
Sofiia Reshetniak, Cristian A. Bogaciu, Stefan Bonn

et al.

The Journal of Physiology, Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 5, 2024

Abstract The synaptic vesicle cluster (SVC) is an essential component of chemical synapses, which provides neurotransmitter‐loaded vesicles during activity, at the same time as also controlling local concentrations numerous exo‐ and endocytosis cofactors. In addition, SVC hosts molecules that participate in other aspects function, from cytoskeletal components to adhesion proteins, affects location function organelles such mitochondria endoplasmic reticulum. We argue here these features extend functional involvement synapse formation, signalling plasticity, well stabilization metabolism. propose changes size coalesce with postsynaptic compartment, supporting interplay between pre‐ dynamics. Thereby, could be seen ‘all‐in‐one’ regulator structure should investigated more detail, reveal molecular mechanisms control heterogeneity. image

Language: Английский

Citations

1
Turning garbage into gold: Autophagy in synaptic function DOI
Erin Smith, Maeve Coughlan, Sandra Maday

et al.

Current Opinion in Neurobiology, Journal Year: 2024, Volume and Issue: 90, P. 102937 - 102937

Published: Dec. 12, 2024

Language: Английский

Citations

1
Absence of ATG9A and synaptophysin demixing on Rab5 mutation-induced giant endosomes DOI Creative Commons
Ji-Young Choi, Yumei Wu, Daehun Park

et al.

Molecular Brain, Journal Year: 2024, Volume and Issue: 17(1)

Published: Sept. 2, 2024

Abstract ATG9A is the only integral membrane protein among core autophagy-related (ATG) proteins. We previously found that does not co-assemble into synaptophysin-positive vesicles, but rather, localizes to a distinct pool of vesicles within synapsin condensates in both fibroblasts and nerve terminals. The endocytic origin these further suggests existence different intracellular sorting or segregation mechanisms for synaptophysin cells. However, precise underlying mechanism remains largely unknown. In this follow-up study, we investigated endosomal localization two proteins by exploiting advantages Rab5 mutant induces formation enlarged endosomes. Notably, intermix perfectly do segregate on giant endosomes, indicating separation solely caused inherent properties proteins, possibly other unknown factors.

Language: Английский

Citations

0
Activity-driven trafficking of endogenous synaptic proteins through proximity labeling DOI Open Access
Carlos Pascual-Caro, Jaime de Juan‐Sanz

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: April 23, 2024

Abstract To enable transmission of information in the brain, synaptic vesicles fuse to presynaptic membranes, liberating their content and exposing transiently a myriad vesicular transmembrane proteins. However, versatile methods for quantifying translocation endogenous proteins during neuronal activity remain unavailable, as fast dynamics vesicle cycling difficult specific isolation trafficking such transient surface exposure. Here we developed novel approach using cleft proximity labeling capture quantify activity-driven at synapse. We show that accelerating biotinylation times match exocytosis allows capturing exposed neural activity, enabling first time study nearly every protein. As proof-of-concept, further applied this technology obtain direct evidence non-canonical proteins, ATG9A NPTX1, which had been proposed traffic but proof not yet shown. The technological advancement presented here will facilitate future studies dissecting molecular identity exocytosed synapse helping define machinery sustains neurotransmission mammalian brain. Significance statement Synaptic is critical neurons communicate sustain brain function. Pascual-Caro de Juan-Sanz develop pioneering method any Coordinating clefts just few seconds, authors visualize This work provides framework uncover complex choreography events occurring within firing synapses, deeper control communication circuit physiology

Language: Английский

Citations

0