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ZFIN ID: ZDB-FIG-130430-33
Blanc et al., 2012 - Critical function for the Ras-GTPase activating protein RASA3 in vertebrate erythropoiesis and megakaryopoiesis. Proceedings of the National Academy of Sciences of the United States of America   109(30):12099-12104 Full text @ Proc. Natl. Acad. Sci. USA
ADDITIONAL FIGURES
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Long-pec to Protruding-mouth

Fig. 4

rasa3 knockdown in zebrafish recapitulates the scat anemia and thrombocytopenia phenotype. (A) In WT embryos, abundant ¿-dianisidine positive hemoglobinized cells are seen in the heart and vessels (arrow) at 72 h postfertilization, whereas positive cells are absent in rasa3 morphants (MO). (B) Quantitation of the range of embryos containing normal and reduced or null number of hemoglobinized cells at 72 h postfertilization from control (uninjected) and rasa3 MO (injected). X ± SEM; *P < 0.05, t test. (C) By RT-PCR, abnormally processed rasa3 mRNA splice forms are found in MO compared with RNA control (CTRL) and uninjected embryos (WT). actb serves as a control of off-target effects. (D) Silencing of rasa3 results in loss of GFP+ cd41-thrombocytes (green) in the transgenic Tg(cd41:GFP) line (MO) compared with control (WT). (E) Quantitation of GFP+ cd41-thrombocytes by flow cytometry in control and rasa3 morphant (MO) embryos. X ± SEM; *P < 0.05, t test. (F) Treatment of WT zebrafish (uninjected) and rasa3 MO (injected) from the Tg(globin LCR:eGFP) transgenic line with the potent antioxidant N-acetyl cysteine (+NAC) fails to improve anemia in the rasa3 MO embryos, indicating that generation of ROS per se is not a contributing factor in the anemia. NAC, exposed to vehicle carrier only. X ± SEM; *P < 0.05; **P < 0.01, t test.

Gene Expression Details No data available
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-rasa3 standard conditions Protruding-mouth erythrocyte differentiation disrupted, abnormal
Protruding-mouth nucleate erythrocyte hemoglobin complex decreased amount, abnormal
WT + MO1-rasa3 + MO2-rasa3 standard conditions Protruding-mouth erythrocyte differentiation disrupted, abnormal
Protruding-mouth nucleate erythrocyte hemoglobin complex decreased amount, abnormal
WT + MO2-rasa3 standard conditions Protruding-mouth erythrocyte differentiation disrupted, abnormal
Protruding-mouth nucleate erythrocyte hemoglobin complex decreased amount, abnormal
la2Tg + MO1-rasa3 standard conditions Long-pec - Protruding-mouth thrombocyte decreased amount, abnormal
Long-pec - Protruding-mouth thrombocyte differentiation disrupted, abnormal
la2Tg + MO1-rasa3 + MO2-rasa3 standard conditions Long-pec - Protruding-mouth thrombocyte decreased amount, abnormal
Long-pec - Protruding-mouth thrombocyte differentiation disrupted, abnormal
la2Tg + MO2-rasa3 standard conditions Long-pec - Protruding-mouth thrombocyte decreased amount, abnormal
Long-pec - Protruding-mouth thrombocyte differentiation disrupted, 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