header logo image header logo text
Downloads Login
Research
General Information
ZIRC
ZFIN ID: ZDB-FIG-070813-8
Hirata et al., 2007 - Zebrafish relatively relaxed mutants have a ryanodine receptor defect, show slow swimming and provide a model of multi-minicore disease. Development (Cambridge, England)   134(15):2771-2781 Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Fish:
Knockdown Reagent:
Anatomical Term:
Stage Range: Prim-5 to Long-pec

Fig. 5 ryr mutant embryos have a mutation in ryr1b due to a DNA insertion that leads to aberrant splicing of ryr1b mRNA. (A) Meiotic mapping placed the ryr locus between z737 and z8343 in chromosome 18. No recombination was found with a polymorphic marker in the ryr1b gene (0/1110). (B) ryr mutants contained a 32-bp insertion (red) in ryr1b mRNA that added a nonsense codon 5′ to the transmembrane domains. (C) Reverse transcriptase (RT)-PCR using primers #1 and #2 (see B) amplified both a longer mutant band (169 bp) and a shorter wild-type band (137 bp) from a group of wild-type siblings (lane 1), whereas the mutant band was predominant in ryr mutants (lane 2). The wild-type band can be seen faintly in the mutant lane. Only the shorter band was amplified from wild-type AB embryos (lane 3). (D) ryr mutants carry a 4046-bp insertion (red) in the intron between exons 48 and 49 of ryr1b that includes the additional 32-bp sequence found in the mutant cDNA. (E) Injection of an antisense morpholino (MO1, see D) into recently fertilized wild-type embryos effectively decreases normal ryr1b mRNA at 24, 36 and 48 hpf, whereas control MO does not. A fragment of normal ryr1b mRNA was examined by RT-PCR with primers #1 and #2 (see D). (F) Genomic PCR and RT-PCR with primers #1 and #2 using individual embryos as a template. Genotypes, as shown, were identified by genomic PCR. Notice that, in mutants, the intensity of the wild-type (short) band amplified by RT-PCR was comparable to that of the mutant (long) band at 24 hpf (lane 3) but was diminished at 48 hpf (lane 6).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
ryr1b WT control Prim-5 whole organism RTPCR
Prim-25 whole organism RTPCR
Long-pec whole organism RTPCR
WT standard conditions Prim-5 whole organism RTPCR
Long-pec whole organism RTPCR
WT + MO1-ryr1b standard conditions Prim-5 whole organism RTPCR
Prim-25 whole organism RTPCR
Long-pec whole organism RTPCR
ryr1bmi340/+ standard conditions Prim-5 whole organism RTPCR
Long-pec whole organism RTPCR
ryr1bmi340/mi340 standard conditions Prim-5 whole organism RTPCR
Long-pec whole organism RTPCR
Antibody Labeling Details No data available
Phenotype Details No data available
Acknowledgments:
ZFIN wishes to thank the journal Development (Cambridge, England) for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Development