γ-Tubulin small complex formation is essential for early zebrafish embryogenesis

Pouchucq, Luis; Undurraga, Cristian; Fuentes, Ricardo; Cornejo, Mauricio; Allende, Miguel L.; Monasterio, Octavio

doi:10.1016/j.mod.2018.06.006

Cited by 3 publications

(5 citation statements)

References 48 publications

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“…Each component of γ-TuSC is likely essential for all cell proliferation in the developing zebrafish. Recently, Pouchucq et al (2018) reported that morpholino (MO)-mediated tubg1 / γ-tubulin knockdown caused zebrafish embryo development arrest at the mid-gastrula stage. The early developmental defects are most probably attributable to the fact that MO was designed against the translation start site of tubg1 mRNA, which blocks the translation of both the maternal and zygotic tubg1 mRNA (Pouchucq et al, 2018).…”

Section: Discussionmentioning

confidence: 99%

“…Recently, Pouchucq et al (2018) reported that morpholino (MO)-mediated tubg1 / γ-tubulin knockdown caused zebrafish embryo development arrest at the mid-gastrula stage. The early developmental defects are most probably attributable to the fact that MO was designed against the translation start site of tubg1 mRNA, which blocks the translation of both the maternal and zygotic tubg1 mRNA (Pouchucq et al, 2018). Although cell cycle defects and increased apoptosis were observed after depletion of γ-tubulin (Pouchucq et al, 2018), study on the function of γ-tubulin in later stages of development has been limited.…”

Section: Discussionmentioning

confidence: 99%

“…The early developmental defects are most probably attributable to the fact that MO was designed against the translation start site of tubg1 mRNA, which blocks the translation of both the maternal and zygotic tubg1 mRNA (Pouchucq et al, 2018). Although cell cycle defects and increased apoptosis were observed after depletion of γ-tubulin (Pouchucq et al, 2018), study on the function of γ-tubulin in later stages of development has been limited. In contrast, the tubgcp3 mutants can survive longer than 10 days due to the maternal deposition of tubgcp3 gene products.…”

Section: Discussionmentioning

confidence: 99%

“…Many studies have shown that γ-TuSC is involved in MT nucleation and mitotic spindle assembly, and every component of γ-TuSC is indispensable for cell cycle progression (Oakley et al, 1990; Geissler et al, 1996; Knop et al, 1997; Yuba-Kubo et al, 2005; Xiong and Oakley, 2009; Pouchucq et al, 2018). In Saccharomyces cerevisiae , cells overexpressing the wild-type Spc98p/GCP3 or carrying the temperature-sensitive allele spc98-1 arrest in mitosis with a defective spindle (Geissler et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

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Tubgcp3 Is Required for Retinal Progenitor Cell Proliferation During Zebrafish Development

Jin

Zhong

2019

Front. Mol. Neurosci.

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The centrosomal protein γ-tubulin complex protein 3 (Tubgcp3/GCP3) is required for the assembly of γ-tubulin small complexes (γ-TuSCs) and γ-tubulin ring complexes (γ-TuRCs), which play critical roles in mitotic spindle formation during mitosis. However, its function in vertebrate embryonic development is unknown. Here, we generated the zebrafish tubgcp3 mutants using the CRISPR/Cas9 system and found that the tubgcp3 mutants exhibited the small eye phenotype. Tubgcp3 is required for the cell cycle progression of retinal progenitor cells (RPCs), and its depletion caused cell cycle arrest in the mitotic (M) phase. The M-phase arrested RPCs exhibited aberrant monopolar spindles and abnormal distributed centrioles and γ-tubulin. Moreover, these RPCs underwent apoptosis finally. Our study provides the in vivo model for the functional study of Tubgcp3 and sheds light on the roles of centrosomal γ-tubulin complexes in vertebrate development.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Tubgcp3 Is Required for Retinal Progenitor Cell Proliferation During Zebrafish Development

Jin

Zhong

2019

Front. Mol. Neurosci.

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“…Microtubules and microtubule-associated proteins such as TUBGCP2 are therefore integral to all fundamental molecular and cellular processes of early development [ 3 , 4 , 9 , 12 , 13 ]. This includes chromosome separation, centrosome, centromere, centriole, and spindle dynamics required for viable symmetric or asymmetric cell division [ 14 , 15 , 16 , 17 , 18 ], cellular proliferation, differentiation, and migration [ 3 , 4 , 19 , 20 , 21 ].…”

Section: Introductionmentioning

confidence: 99%

A Zebrafish/Drosophila Dual System Model for Investigating Human Microcephaly

Bartoszewski

Dawidziuk²,

Kasica

et al. 2022

Cells

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Microcephaly presents in neurodevelopmental disorders with multiple aetiologies, including bi-allelic mutation in TUBGCP2, a component of the biologically fundamental and conserved microtubule-nucleation complex, γ-TuRC. Elucidating underlying principles driving microcephaly requires clear phenotype recapitulation and assay reproducibility, areas where go-to experimental models fall short. We present an alternative simple vertebrate/invertebrate dual system to investigate fundamental TUBGCP2-related processes driving human microcephaly and associated developmental traits. We show that antisense morpholino knockdown (KD) of the Danio rerio homolog, tubgcp2, recapitulates human TUBGCP2-associated microcephaly. Co-injection of wild type mRNA pre-empts microcephaly in 55% of KD zebrafish larvae, confirming causality. Body shortening observed in morphants is also rescued. Mitotic marker (pH3) staining further reveals aberrantly accumulated dividing brain cells in microcephalic tubgcp2 KD morphants, indicating that tubgcp2 depletion disrupts normal mitosis and/or proliferation in zebrafish neural progenitor brain cells. Drosophila melanogaster double knockouts (KO) for TUBGCP2 homologs Grip84/cg7716 also develop microcephalic brains with general microsomia. Exacerbated Grip84/cg7716-linked developmental aberration versus single mutations strongly suggests interactive or coinciding gene functions. We infer that tubgcp2 and Grip84/cg7716 affect brain size similarly to TUBGCP2 and recapitulate both microcephaly and microcephaly-associated developmental impact, validating the zebrafish/fly research model for human microcephaly. Given the conserved cross-phyla homolog function, the data also strongly support mitotic and/or proliferative disruption linked to aberrant microtubule nucleation in progenitor brain cells as key mechanistic defects for human microcephaly.

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Tubgcp3 is a mitotic regulator of planarian epidermal differentiation

Lin

Zhang

et al. 2021

Gene

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γ-Tubulin small complex formation is essential for early zebrafish embryogenesis

Cited by 3 publications

References 48 publications

Tubgcp3 Is Required for Retinal Progenitor Cell Proliferation During Zebrafish Development

Tubgcp3 Is Required for Retinal Progenitor Cell Proliferation During Zebrafish Development

A Zebrafish/Drosophila Dual System Model for Investigating Human Microcephaly

Tubgcp3 is a mitotic regulator of planarian epidermal differentiation

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