ZFIN ID: ZDB-FIG-150819-2
Gao et al., 2015 - Mecp2 regulates neural cell differentiation by suppressing the Id1-Her2/Hes5 axis in zebrafish. Journal of Cell Science   128(12):2340-50 Full text @ J. Cell Sci.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage: Long-pec
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: Long-pec

Fig. 2

Knockdown of mecp2 increases neural precursors and astrogenesis while suppressing neurogenesis. (A,A2) RNA in situ hybridization performed with nestin probe in controls (wild-type, WT) (A) and mecp2 morphants (A′) at 48hpf. Note the upregulated nestin expression in the developing brain of mecp2 morphants. (B–E′) mecp2-MOs were injected into Tg(ef1α:mAG-zGem) transgenic embryos, in which GFP-positive cells are in the S, G2 and M phase of the cell cycle. Immunostaining was performed using an anti-pH3 antibody (red). The numbers of green fluorescence-positive cells and pH3-positive mitotic cells were increased in mecp2 morphants at 48hpf (C′,D′,E′) compared with the wild-type (C,D,E). Numbers of proliferative cells labeled by Tg(ef1α:mAG-zGem) were counted using Imaris software (B). Dorsal views are shown with anterior to the left (A,A&prime& C–E′). (F–H′) RNA in situ hybridization revealed that gfap expression was increased in the developing brain of mecp2 morphants (F′), compared with the wild-type (F) at 48hpf. However, map2 and neurod expression was reduced in mecp2 morphants (G′,H′) compared with controls (G,H) at 48hpf. (I,I′) mecp2-MOs were injected into Tg(neurod:EGFP) transgenic embryos. Note that neurod was downregulated in mecp2 morphants (I′), compared with controls (I) at 48hpf. (J,J′) Whole-mount immunostaining with antibody against Huc. Huc was downregulated in mecp2 morphants (J′), compared with controls (J) at 48hpf. (K,K′) mecp2-MOs were injected into Tg(islet1:EGFP) transgenic embryos, in which GFP-positive cells are motor neurons. The number of GFP-positive cells was reduced in mecp2 morphants (K′), compared with the wild-type (K) at 48hpf. Dorsal views are shown with anterior to the left (F–K′). (L) Quantitative real-time PCR analysis of related genes in wild-type embryos and mecp2 morphants, normalized to gapdh. Measurements are the mean±s.d. from three independent experiments. *P<0.05. In A,A′, and C–K′, numbers in the bottom right corner represent the number of embryos showing the indicated phenotype/total embryos examined.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP nl1Tg standard conditions Long-pec cerebellum neuron IHC
Long-pec eye IHC
Long-pec inner ear IHC
Long-pec olfactory bulb neuron IHC
nl1Tg + MO2-mecp2 standard conditions Long-pec cerebellum neuron IHC
Long-pec eye IHC
Long-pec inner ear IHC
Long-pec olfactory bulb neuron IHC
elavl3 WT standard conditions Long-pec brain RTPCR
WT + MO2-mecp2 standard conditions Long-pec brain RTPCR
Ab1-elavl nl1Tg standard conditions Long-pec brain neuron IHC
Ab1-elavl nl1Tg + MO2-mecp2 standard conditions Long-pec brain neuron IHC
gfap WT standard conditions Long-pec brain RTPCR
Long-pec brain neuronal stem cell ISH
Long-pec brain radial glial cell ISH
Long-pec forebrain ISH
Long-pec hindbrain ISH
Long-pec midbrain ISH
WT + MO2-mecp2 standard conditions Long-pec brain RTPCR
Long-pec brain neuronal stem cell ISH
Long-pec brain radial glial cell ISH
Long-pec forebrain ISH
Long-pec hindbrain ISH
Long-pec midbrain ISH
GFP rw0Tg standard conditions Long-pec facial ganglion IHC
Long-pec motor neuron IHC
Long-pec trigeminal ganglion IHC
Long-pec vagal ganglion IHC
rw0Tg + MO2-mecp2 standard conditions Long-pec facial ganglion IHC
Long-pec motor neuron IHC
Long-pec trigeminal ganglion IHC
Long-pec vagal ganglion IHC
isl1 WT standard conditions Long-pec brain RTPCR
WT + MO2-mecp2 standard conditions Long-pec brain RTPCR
mAGFP pku322Tg standard conditions Long-pec brain neuronal stem cell IHC
pku322Tg + MO2-mecp2 standard conditions Long-pec brain neuronal stem cell IHC
map2 WT standard conditions Long-pec brain RTPCR
Long-pec brain neuron ISH
WT + MO2-mecp2 standard conditions Long-pec brain RTPCR
Long-pec brain neuron ISH
nes WT standard conditions Long-pec brain RTPCR
Long-pec facial ganglion ISH
Long-pec hindbrain ISH
Long-pec lateral line ganglion ISH
Long-pec midbrain hindbrain boundary posterior region ISH
Long-pec neuronal stem cell ISH
Long-pec pretectum ISH
Long-pec statoacoustic (VIII) ganglion ISH
Long-pec vagal ganglion ISH
Long-pec ventral telencephalon ISH
WT + MO2-mecp2 standard conditions Long-pec brain RTPCR
Long-pec facial ganglion ISH
Long-pec hindbrain ISH
Long-pec lateral line ganglion ISH
Long-pec midbrain hindbrain boundary posterior region ISH
Long-pec neuronal stem cell ISH
Long-pec pretectum ISH
Long-pec statoacoustic (VIII) ganglion ISH
Long-pec vagal ganglion ISH
Long-pec ventral telencephalon ISH
neurod1 WT standard conditions Long-pec brain RTPCR
Long-pec cerebellum neuron ISH
Long-pec eye ISH
Long-pec inner ear ISH
Long-pec olfactory bulb neuron ISH
WT + MO2-mecp2 standard conditions Long-pec brain RTPCR
Long-pec cerebellum neuron ISH
Long-pec eye ISH
Long-pec inner ear ISH
Long-pec olfactory bulb neuron ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO2-mecp2 standard conditions Long-pec brain has extra parts of type astrocyte, abnormal
Long-pec brain has extra parts of type neuronal stem cell, abnormal
Long-pec brain has fewer parts of type neuron, abnormal
Long-pec brain astrocyte differentiation increased occurrence, abnormal
Long-pec brain neural precursor cell proliferation increased occurrence, abnormal
Long-pec brain neuron differentiation decreased occurrence, abnormal
Long-pec brain neuronal stem cell proliferative, abnormal
nl1Tg + MO2-mecp2 standard conditions Long-pec brain has fewer parts of type neuron, abnormal
Long-pec brain neuron differentiation decreased occurrence, abnormal
pku322Tg + MO2-mecp2 standard conditions Long-pec brain neural precursor cell proliferation increased occurrence, abnormal
Long-pec brain neuronal stem cell proliferative, abnormal
rw0Tg + MO2-mecp2 standard conditions Long-pec brain neuron differentiation decreased occurrence, abnormal
Long-pec facial ganglion has fewer parts of type motor neuron, abnormal
Long-pec trigeminal ganglion has fewer parts of type motor neuron, abnormal
Long-pec vagal ganglion has fewer parts of type motor neuron, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Journal of Cell Science for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ J. Cell Sci.