Growth cone advance requires EB1 as revealed by genomic replacement with a light-sensitive variant.
EB1
cell biology
growth cone
human
induced pluripotent stem cells
microtubules
neuron morphogenesis
neuroscience
optogenetics
Journal
eLife
ISSN: 2050-084X
Titre abrégé: Elife
Pays: England
ID NLM: 101579614
Informations de publication
Date de publication:
30 01 2023
30 01 2023
Historique:
received:
12
10
2022
accepted:
27
01
2023
pubmed:
31
1
2023
medline:
15
2
2023
entrez:
30
1
2023
Statut:
epublish
Résumé
A challenge in analyzing dynamic intracellular cell biological processes is the dearth of methodologies that are sufficiently fast and specific to perturb intracellular protein activities. We previously developed a light-sensitive variant of the microtubule plus end-tracking protein EB1 by inserting a blue light-controlled protein dimerization module between functional domains. Here, we describe an advanced method to replace endogenous EB1 with this light-sensitive variant in a single genome editing step, thereby enabling this approach in human induced pluripotent stem cells (hiPSCs) and hiPSC-derived neurons. We demonstrate that acute and local optogenetic EB1 inactivation in developing cortical neurons induces microtubule depolymerization in the growth cone periphery and subsequent neurite retraction. In addition, advancing growth cones are repelled from areas of blue light exposure. These phenotypes were independent of the neuronal EB1 homolog EB3, revealing a direct dynamic role of EB1-mediated microtubule plus end interactions in neuron morphogenesis and neurite guidance.
Identifiants
pubmed: 36715499
doi: 10.7554/eLife.84143
pii: 84143
pmc: PMC9917429
doi:
pii:
Substances chimiques
Microtubule-Associated Proteins
0
MAPRE1 protein, human
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : NCRR NIH HHS
ID : S10 RR026758
Pays : United States
Organisme : NINDS NIH HHS
ID : R01 NS107480
Pays : United States
Organisme : NIH HHS
ID : S10 OD028611
Pays : United States
Organisme : NCI NIH HHS
ID : R21 CA224194
Pays : United States
Organisme : NCI NIH HHS
ID : R21 CA224194
Pays : United States
Organisme : NINDS NIH HHS
ID : R01 NS107480
Pays : United States
Organisme : NIH HHS
ID : S10 RR026758
Pays : United States
Organisme : NIH HHS
ID : S10 OD028611
Pays : United States
Informations de copyright
© 2023, Dema et al.
Déclaration de conflit d'intérêts
AD, RC, SR, Jv, TW No competing interests declared
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