ZFIN ID: ZDB-FIG-090915-51
Calarco et al., 2009 - Regulation of vertebrate nervous system alternative splicing and development by an SR-related protein. Cell   138(5):898-910 Full text @ Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: Prim-5 to Long-pec
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Prim-15 to Long-pec

Fig. 6 nSR100 Regulates Vertebrate Nervous System and Sensory Organ Development

(A–C) RNA in situ hybridization assays monitoring ZnSR100 expression in the developing zebrafish embryo.

(A and A′) expression at 24 hpf, lateral and frontal views, respectively.

(B and B′) nSR100 expression at 36 hpf, lateral and dorsal views, respectively.

(C and C′) nSR100 expression at 48 hpf, lateral and dorsal views, respectively.

(D) p53 morpholino antisense oligonucleotide (MO) control-injected embryo at 50 hpf.

(E) p53 + ZnSR100 MOspl-injected embryo at 50 hpf.

(F) p53 + ZnSR100 MOspl-injected embryo at 50 hpf, rescued through coinjection of ZnSR100 mRNA.

(G–I) RNA in situ hybridization assays monitoring islet-1 expression in 50 hpf embryos. p53-MO-injected control (G), nSR100spl morphant embryo (H), and nSR100spl morphant embryo rescued through coinjection of 20 pg ZnSR100 mRNA (I) are shown.

(J–M) Confocal projections of trigeminal ganglion in Tg(sensory:GFP) transgenic embryos. Control embryo at 30 hpf (J), representative nSR100spl morphant embryo at 30 hpf (K), control embryo at 36 hpf (L), and representative nSR100spl morphant embryo at 36 hpf (M) are shown.

(N) nSR100-regulated alternative exons conserved between zebrafish and mouse display altered splicing patterns in nSR100spl morphant embryos, relative to control embryos. Total RNA was isolated from the heads of wild-type and nSR100spl morphant embryos at 50 hpf, and semiquantitative RT-PCR was performed to analyze AS patterns. Percent exon inclusion values are shown below each lane.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP zf154Tg + MO1-srrm4 + MO4-tp53 standard conditions Prim-15 trigeminal ganglion axon IFL
Prim-25 trigeminal ganglion axon IFL
zf154Tg + MO4-tp53 standard conditions Prim-15 trigeminal ganglion axon IFL
Prim-25 trigeminal ganglion axon IFL
isl1a AB/TL + MO1-srrm4 + MO4-tp53 standard conditions Long-pec brain ISH
Long-pec eye ISH
AB/TL + MO4-tp53 standard conditions Long-pec brain ISH
Long-pec eye ISH
srrm4 AB/TL standard conditions Prim-5 hindbrain ISH
Prim-5 midbrain hindbrain boundary ISH
Prim-5 optic tectum ISH
Prim-5 telencephalon ISH
Prim-5 trigeminal ganglion ISH
Prim-25 cerebellum ISH
Prim-25 hindbrain ISH
Prim-25 midbrain hindbrain boundary ISH
Prim-25 optic tectum ISH
Prim-25 retina ISH
Prim-25 telencephalon ISH
Long-pec brain ISH
Long-pec eye ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
AB/TL + MO1-srrm4 + MO4-tp53 standard conditions Long-pec ventricular system increased volume, abnormal
Long-pec whole organism curved ventral, abnormal
zf154Tg + MO1-srrm4 + MO4-tp53 standard conditions Prim-15 axonogenesis disrupted, abnormal
Prim-15 trigeminal ganglion axon decreased length, abnormal
Prim-25 axonogenesis disrupted, abnormal
Prim-25 trigeminal ganglion axon decreased length, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Cell for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Cell, 138(5), Calarco, J.A., Superina, S., O'Hanlon, D., Gabut, M., Raj, B., Pan, Q., Skalska, U., Clarke, L., Gelinas, D., van der Kooy, D., Zhen, M., Ciruna, B., and Blencowe, B.J., Regulation of vertebrate nervous system alternative splicing and development by an SR-related protein, 898-910, Copyright (2009) with permission from Elsevier. Full text @ Cell