PXD024177

PXD024177 is an original dataset announced via ProteomeXchange.

Title	Quantitative profiling of axonal guidance proteins during the differentiation of human neurospheres
Description	Axon guidance is required for the establishment of brain circuits. Whether much of the molecular basis of axon guidance is known from animal models, the molecular machinery coordinating axon growth and pathfinding in humans remains to be elucidated. The use of induced pluripotent stem cells (iPSC) from human donors has revolutionized in vitro studies of the human brain. iPSC can be differentiated into neuronal stem cells which can be used to generate neural tissue-like cultures, known as neurospheres, that reproduce, in many aspects, the cell types and molecules present in the brain. Here, we analyzed quantitative changes in the proteome of neurospheres during differentiation. Relative quantification was performed at early time points during differentiation using iTRAQ-based labeling and LC-MS/MS analysis. We identified 6438 proteins, from which 433 were downregulated and 479 were upregulated during differentiation. We show that human neurospheres have a molecular profile that correlates to the fetal brain. During differentiation, upregulated pathways are related to neuronal development and differentiation, cell adhesion and axonal guidance whereas cell proliferation pathways were downregulated. We developed a functional assay to check for neurite outgrowth in neurospheres and confirmed that neurite outgrowth potential is increased after 10 days of differentiation and is enhanced by increasing cyclic AMP levels. The proteins identified here represent a resource to monitor neurosphere differentiation and coupled to the neurite outgrowth assay can be used to functionally explore neurological disorders using human neurospheres as a model.
HostingRepository	PRIDE
AnnounceDate	2021-02-25
AnnouncementXML	Submission_2021-02-24_22:16:20.753.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Michele Martins
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	iTRAQ4plex-116 reporter+balance reagent acylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2021-02-15 03:22:16	ID requested
⏵ 1	2021-02-24 22:16:21	announced

Dataset with its publication pending

submitter keyword: iTRAQ-based quantitative proteomics, human induced pluripotent stem cells , neural stem cells (NSC), human neurospheres, neural differentiation, axon guidance

Magno Junqueira
contact affiliation	Proteomics Unit, Department of Biochemistry, Chemistry Institute, Federal University of Rio de Janeiro (UFRJ). Av. Athos da Silveira Ramos, 149 - bloco A 5° andar - Cidade Universitária da Universidade Federal do Rio de Janeiro, Rio de Janeiro - RJ, 21941-909, Brazil.
contact email	magnojunqueira@iq.ufrj.br
lab head
Michele Martins
contact affiliation	UFRJ
contact email	miih.martiins@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD024177
     1. Label: PRIDE project
     2. Name: Quantitative profiling of axonal guidance proteins during the differentiation of human neurospheres