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Dev Biol 2015 Dec 15;4082:269-91. doi: 10.1016/j.ydbio.2015.03.010.
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An in vivo screen to identify candidate neurogenic genes in the developing Xenopus visual system.

Bestman JE , Huang LC , Lee-Osbourne J , Cheung P , Cline HT .

Neurogenesis in the brain of Xenopus laevis continues throughout larval stages of development. We developed a 2-tier screen to identify candidate genes controlling neurogenesis in Xenopus optic tectum in vivo. First, microarray and NanoString analyses were used to identify candidate genes that were differentially expressed in Sox2-expressing neural progenitor cells or their neuronal progeny. Then an in vivo, time-lapse imaging-based screen was used to test whether morpholinos against 34 candidate genes altered neural progenitor cell proliferation or neuronal differentiation over 3 days in the optic tectum of intact Xenopus tadpoles. We co-electroporated antisense morpholino oligonucleotides against each of the candidate genes with a plasmid that drives GFP expression in Sox2-expressing neural progenitor cells and quantified the effects of morpholinos on neurogenesis. Of the 34 morpholinos tested, 24 altered neural progenitor cell proliferation or neuronal differentiation. The candidates which were tagged as differentially expressed and validated by the in vivo imaging screen include: actn1, arl9, eif3a, elk4, ephb1, fmr1-a, fxr1-1, fbxw7, fgf2, gstp1, hat1, hspa5, lsm6, mecp2, mmp9, and prkaca. Several of these candidates, including fgf2 and elk4, have known or proposed neurogenic functions, thereby validating our strategy to identify candidates. Genes with no previously demonstrated neurogenic functions, gstp1, hspa5 and lsm6, were identified from the morpholino experiments, suggesting that our screen successfully revealed unknown candidates. Genes that are associated with human disease, such as such as mecp2 and fmr1-a, were identified by our screen, providing the groundwork for using Xenopus as an experimental system to probe conserved disease mechanisms. Together the data identify candidate neurogenic regulatory genes and demonstrate that Xenopus is an effective experimental animal to identify and characterize genes that regulate neural progenitor cell proliferation and differentiation in vivo.

PubMed ID: 25818835
PMC ID: PMC4584193
Article link: Dev Biol
Grant support: [+]

Species referenced: Xenopus laevis
Genes referenced: actn1 arl9 armc8 chn2 cpeb1 ctdnep1 efna3 eif3a elk4 ephb1 epx fbxw7 fgf2 fmr1 fxr1 glis2 gstp1 hat1 hdac6 hspa5 hspd1 lsm6 mecp2 mkrn2 mocs3 prkaca pura r3hdm2 rbfox2 slc12a2 sox2 tecta.2 tle1 vangl1 wnt7b
Morpholinos: actn1 MO1 arl9 MO1 armc8 MO1 chn1 MO1 cpeb1 MO2 ctdnep1 MO3 dio3 MO1 efna3 MO1 eif3a MO1 elk4 MO1 ephb1 MO1 epx MO1 fbxw7 MO1 fgf2 MO1 fmr1 MO2 fxr1 MO1 glis2 MO1 gstp1 MO1 hat1 MO1 hdac6 MO1 hspa5 MO2 lsm6 MO1 mecp2 MO2 mkrn2 MO3 mmp9.1 MO2 mocs3 MO1 prkaca MO2 pura MO1 r3hdm2 MO1 rbfox2 MO1 slc12a2 MO1 tle1 MO1 vangl1 MO1 wnt7b MO1

Phenotypes: Xla Wt + actn1 MO (fig.8) [+]

Article Images: [+] show captions
References [+] :
Ageta-Ishihara, Septins promote dendrite and axon development by negatively regulating microtubule stability via HDAC6-mediated deacetylation. 2013, Pubmed