Zn2+ decoration of microtubules arrests axonal transport and displaces tau, doublecortin, and MAP2C

The Journal of Cell Biology, Год журнала: 2023, Номер 222(8)

Опубликована: Июнь 16, 2023

Intracellular Zn2+ concentrations increase via depolarization-mediated influx or intracellular release, but the immediate effects of signals on neuron function are not fully understood. By simultaneous recording cytosolic and organelle motility, we find that elevated (IC50 ≈ 5–10 nM) reduces both lysosomal mitochondrial motility in primary rat hippocampal neurons HeLa cells. Using live-cell confocal microscopy vitro single-molecule TIRF imaging, reveal inhibits activity motor proteins (kinesin dynein) without disrupting their microtubule binding. Instead, directly binds to microtubules selectively promotes detachment tau, DCX, MAP2C, MAP1B, MAP4, MAP7, MAP9, p150glued. Bioinformatic predictions structural modeling show binding sites partially overlap with dynein, kinesin. Our results intraneuronal regulates axonal transport microtubule-based processes by interacting microtubules.

Язык: Английский

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High-throughput assay for regulated secretion of neuropeptides in mouse and human neurons

Journal of Biological Chemistry, Год журнала: 2024, Номер 300(6), С. 107321 - 107321

Опубликована: Апрель 25, 2024

Язык: Английский

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Myosin VI controls localization of Golgi satellites at active presynaptic boutons

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

Опубликована: Май 9, 2024

Abstract Neurons, as long-living non-dividing cells with complex morphology, depend on highly elaborate secretory trafficking system which ensures the constant delivery, removal and recycling of proteins membranes. Previously, we have shown that simplified Golgi-related structures called Golgi satellites (GS), distinct from somatic complex, are present in dendrites primary hippocampal neurons involved glycosylation local forward membrane proteins. However, whether GS also targeted to axons principal not been explored. Here, investigate subcellular distribution adult discover mobile stationary along entire axonal length, extending distal tips growth cone. Live imaging experiments revealed neuronal firing modulates switch between long range transport mediated by kinesin dynein stalling. We found frequently pause or stop at pre-synaptic sites activity-dependent manner. This behavior depends actin cytoskeleton actin-based motor protein myosin VI. Overall, our study demonstrates activity can dynamically regulate positioning axon, shedding light intricate mechanisms underlying organelle neurons. Significance statement Our unveils presence dynamic specialized organelles implicated trafficking, within throughout including cone regions. transported bidirectionally their preferential pausing presynaptic is regulated firing. boutons relies VI protein. work elucidates how distribution, fundamental

Язык: Английский

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Rab10 regulates neuropeptide release by maintaining Ca2+ homeostasis and protein synthesis

Опубликована: Март 19, 2024

Dense core vesicles (DCVs) transport and release various neuropeptides neurotrophins that control diverse brain functions, but the DCV secretory pathway remains poorly understood. Here, we tested a prediction emerging from invertebrate studies about crucial role of intracellular trafficking GTPase Rab10, by assessing exocytosis at single- cell resolution upon acute Rab10 depletion in mature mouse hippocampal neurons, to circumvent potential confounding effects Rab10’s established neurite outgrowth.We observed significant inhibition Rab10-depleted whereas synaptic vesicle was unaffected. However, rather than direct involvement trafficking, this effect attributed two ER-dependent processes, ER-regulated Ca 2+ dynamics protein synthesis. Gene ontology analysis differentially expressed proteins identified substantial alterations ER/ribosomal proteins, including -pump SERCA2. In addition, ER morphology were altered, levels depleted homeostasis impaired neurons. entry using ionophore still triggered less exocytosis. Instead, leucine supplementation, which enhances synthesis, largely rescued deficiency. We conclude is required for neuropeptide maintaining regulating Furthermore, appeared more dependent on (acute) synthesis

Язык: Английский

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SNARE protein tomosyn regulates dense core vesicle composition but not exocytosis in mammalian neurons

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2022, Номер unknown

Опубликована: Дек. 19, 2022

Abstract Tomosyn is a large, non-canonical SNARE protein proposed to act as competitive inhibitor of complex formation in vesicle exocytosis. In the brain, tomosyn inhibits fusion synaptic vesicles (SVs), whereas its role neuropeptide-containing dense core (DCVs) unknown. Here, we addressed this question using new mouse model allowing conditional deletion ( Stxbp5 ) and paralogue tomosyn-2 Stxbp5l ), an assay that detects DCV exocytosis with single resolution primary hippocampal neurons. Surprisingly, loss both tomosyns did not affect but resulted strong reduction intracellular levels many cargos, most prominently brain-derived neurotrophic factor (BDNF), granin VGF prohormone convertase PCSK1. Reduced cargos were paralleled by decreased size impaired mRNA expression corresponding genes. We conclude regulate neuropeptide neurotrophin secretion via control cargo production, at step release. Our findings suggest differential effect on two main secretory pathways mammalian neurons argues against conserved formation.

Язык: Английский

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