ZFIN ID: ZDB-FIG-160311-1
Fernández-Murray et al., 2016 - Glycine and Folate Ameliorate Models of Congenital Sideroblastic Anemia. PLoS Genetics   12:e1005783 Full text @ PLoS Genet.
Anatomical Terms:
Stage Range: Prim-5 to Prim-15
Knockdown Reagents:
Observed In:
Stage: Long-pec

Fig. 6

Rescue of the zebrafish model of congenital sideroblastic anemia.A) Whole-mount in situ hybridizations with probes for slc25a38a and slc25a38b at 24 and 34 hpf stages of development demonstrate predominant erythroid expression. For each stage and probe, head and tail views are shown and sites of expression are labeled. Abbreviations: pbi-posterior blood island, pcv-posterior cardinal vein, ss-somites, bl-blood, ret-retina, br-brain, h-heart. Expression of slc25a38b was observed in the posterior blood island, blood and posterior cardinal vein consistent with its preferential expression in erythroid progenitors and erythrocytes, whereas slc25a38a had the same blood-related expression pattern features and was also expressed in retina, brain and somites at 24 hpf. B) Representative images for hemoglobin staining using o-dianisidine of 48 hpf zebrafish embryos injected with slc25a38a+b morpholinos, or standard control morpholino (STD MO), treated with 100 mM glycine, 1 mM sodium folate, or both, starting from 4 hours post-injection. To control for morpholino specificity, we also injected 5-mismatch (5MM) versions of the same slc25a38a+b morpholinos and stained the injected embryos with o-dianisidine. For STD MO and 5MM morphants, images from only the STD MO untreated group are presented because of the low variation in o-dianisidine scores of untreated embryos versus those where glycine and/or folate were present. C) Graph of the o-dianisidine staining heme scores of 48 hpf zebrafish embryos injected with either slc25a38a+b morpholinos (slcMO) or STD MO and then treated from 4 through to 48 hpf with glycine, folate, or both glycine plus folate. All embryos were scored as having “low”, “medium” or “normal” hemoglobin levels in a blinded manner based on visual inspection of o-dianisidine staining. The total numbers (right axis) of scored embryos are indicated, from a total of three independent experiments. The “***” indicates that the p-value between the numbers of embryos in different scoring categories for slc25a38a+b morphants treated with glycine and folate versus the untreated slc25a38a+b morphant group is <0.001.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
slc25a38a WT standard conditions Prim-5 blood ISH
Prim-5 blood island ISH
Prim-5 brain ISH
Prim-5 optic cup ISH
Prim-5 posterior cardinal vein ISH
Prim-5 somite ISH
Prim-15 blood ISH
slc25a38b WT standard conditions Prim-5 blood ISH
Prim-5 blood island ISH
Prim-5 posterior cardinal vein ISH
Prim-15 blood ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
mitfaw2/w2; mpv17a9/a9 + MO1-slc25a38a + MO1-slc25a38b standard conditions Long-pec erythroblast increased amount, abnormal
Long-pec nucleate erythrocyte hemoglobin complex decreased amount, abnormal
mitfaw2/w2; mpv17a9/a9 + MO1-slc25a38a + MO1-slc25a38b chemical treatment: glycine, chemical treatment: folic acid Long-pec nucleate erythrocyte hemoglobin complex normal amount, ameliorated
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