PXD072589

PXD072589 is an original dataset announced via ProteomeXchange.

Title	Modelling Synaptic Maturation from Growth Cone to Synapse in Human Organoids
Description	Human neural organoids (NOs) provide a powerful platform for investigating synaptic development and dysfunction during early neurodevelopment. However, methodologies for isolating functional synaptic structures from these models remain limited. Here, we present a differential centrifugation protocol enabling the enrichment of growth cone particles (GCPs) and immature synaptosomes from air-liquid interface cerebral organoids (ALI-COs) at distinct developmental stages (day 90 and 150). Notably, the method avoids density gradients, requires minimal starting material while maintaining reproducibility across human and murine tissues. Quantitative proteomic profiling revealed significant enrichment of growth cone markers (e.g. GAP43) and classical synaptosomal proteins (e.g. PCLO, BSN, SYN1). Transmission electron microscopy (TEM) confirmed the presence of membrane-enclosed GCPs with fibrous content and mitochondria in day 90 isolates, and immature synaptosomes containing synaptic vesicles on day 150. Functional viability of both types of synaptic structures was demonstrated through KCl-induced depolarization, which triggered phosphorylation changes in growth cone proteins (GAP43, MARCKS, MARCKSL1), cytoskeletal regulators (DCLK1, SHTN1, MARK4, MAP1B) and protein kinases (CAMK2G, PRKCE) in day 90 GCPs, as well as classical synaptic vesicle cycle proteins (SYN1, DNM1, RPH3A) at day 150. Overall, this study establishes a centrifugation-based protocol for isolating growth cones and immature synapses from human organoids, capturing key stages of synaptic development and enabling scalable, patient-compatible models to study synaptic function and dysfunction in neurodevelopmental and neurodegenerative disorders.
HostingRepository	PRIDE
AnnounceDate	2026-05-25
AnnouncementXML	Submission_2026-05-25_02:03:00.437.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Marie Sejberg Øhlenschlæger
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606; scientific name: Mus musculus (Mouse); NCBI TaxID: NEWT:10090;
ModificationList	phosphorylated residue
Instrument	Orbitrap Eclipse; timsTOF Pro

Revision	Datetime	Status	ChangeLog Entry
0	2026-01-02 16:00:52	ID requested
⏵ 1	2026-05-25 02:03:01	announced

10.1111/jnc.70458;

Ø, hlenschl, æ, ger MS, Criscuolo L, Jensen P, Lloyd-Davies S, á, nchez DJ, Sutcliffe M, Bhosale S, Bogetofte H, Tahir M, Jakobsen LA, Pihl M, Brewer J, Schw, ä, mmle V, Poulsen FR, Freude K, Lancaster MA, Robinson PJ, Larsen MR, Modeling Synaptic Maturation From Growth Cone to Synapse in Human Organoids. J Neurochem, 170(5):e70458(2026) [pubmed]

submitter keyword: synapse, synaptosomes, guided dorsal forebrain organoid, stimulation, nerve terminals, Dynamin., subcellular enrichment, proteomics, neural organoid,Brain organoid, GAP43, phosphorylation, air-liquid interface cerebral organoid, growth cone, differential centrifugation

Martin Røssel Larsen
contact affiliation	Department of Biochemistry and Molecular Biology, University of Southern Denmark, Denmark
contact email	mrl@bmb.sdu.dk
lab head
Marie Sejberg Øhlenschlæger
contact affiliation	SDU
contact email	masoe@bmb.sdu.dk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD072589
     1. Label: PRIDE project
     2. Name: Modelling Synaptic Maturation from Growth Cone to Synapse in Human Organoids