ZFIN ID: ZDB-FIG-180209-1
Botthof et al., 2017 - Loss of the homologous recombination gene rad51 leads to Fanconi anemia-like symptoms in zebrafish.. Proceedings of the National Academy of Sciences of the United States of America   114(22):E4452-E4461 Full text @ Proc. Natl. Acad. Sci. USA
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibodies:
Fish:
Anatomical Terms:
Stage Range: 1-cell to Adult
PHENOTYPE:
Fish:
Conditions:
Observed In:
Stage Range: Prim-5 to Long-pec

Fig. 1

The rad51sa23805 allele leads to loss of Rad51 protein and causes DNA damage sensitivity. (A) Western blot showing the expression of Rad51 in testes extracts of WT and mutant zebrafish. β-Actin was used as a loading control. (B) Representative Rad51 immunostained embryos derived from a rad51+/− in-cross. Secondary only (i), Abcam primary (ii), AnaSpec primary (iii). (Magnification, 65×.) (C) Chromosome spreads of 24-hpf WT and mutant embryos treated with 1 µg/mL DEB for 20 h taken using a 100× oil-immersion objective. White arrows indicate characteristic damage (chromosome breaks and radial structures) in response to cross-linking agents (i). Quantification of the damage (ii). Mann–Whitney test, P = 0.0004, n+/+ = 25, n−/− = 26. (D) Comparison of the response of 48-hpf WT and mutant embryos to irradiation (i). (Magnification, 15×.) The black arrow indicates the small head and eye phenotype, which is quantified in ii. Two-tailed Fisher’s exact test pooling WT and heterozygotes as control group, P < 0.001, n = 67. (E) Immunostaining for pH2AX in WT and mutant embryos with pictures of representative embryos taken with a 63× water-immersion objective (i) and quantification of foci (ii). White arrows indicate example foci. Two-tailed Student’s t test, P < 0.0001, n+/+ = 8, n−/− = 11.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
rad51 WT standard conditions Adult testis WB
Ab3-rad51 rad51sa23805/sa23805 standard conditions 1-cell whole organism IHC
Adult Not Detected testis WB
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab2-rad51 IHC rad51sa23805/sa23805 standard conditions 1-cell whole organism
Phenotype Details
Fish Conditions Stage Phenotype
rad51sa23805/sa23805 chemical treatment: diepoxybutane Prim-5 whole organism chromosome damaged, abnormal
rad51sa23805/sa23805 gamma ray Long-pec eye decreased size, abnormal
Long-pec head decreased size, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Proceedings of the National Academy of Sciences of the United States of America for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Proc. Natl. Acad. Sci. USA