- Title
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ZRANB3 is an African-specific type 2 diabetes locus associated with beta-cell mass and insulin response
- Authors
- Adeyemo, A.A., Zaghloul, N.A., Chen, G., Doumatey, A.P., Leitch, C.C., Hostelley, T.L., Nesmith, J.E., Zhou, J., Bentley, A.R., Shriner, D., Fasanmade, O., Okafor, G., Eghan, B., Agyenim-Boateng, K., Chandrasekharappa, S., Adeleye, J., Balogun, W., Owusu, S., Amoah, A., Acheampong, J., Johnson, T., Oli, J., Adebamowo, C., South Africa Zulu Type 2 Diabetes Case-Control Study, Collins, F., Dunston, G., Rotimi, C.N.
- Source
- Full text @ Nat. Commun.
Generation of zranb3 genomic zebrafish mutants. a, b Identified mutations found in homozygous F2 animals by CRISPR/Cas9 in Tg(ins:mCherry) or wild-type animals. In both lines genomic DNA deletions in exon 4 resulted in frame shift and premature stop codons (red box), confirmed by sequencing. c Confocal microscopy images of ins:mCherry transgene (red) in principal islets of wild type or mutant 5 dpf larvae. d Confocal images of whole-mount immunostained islets of wild type or mutant 5 dpf larvae, using antibodies against insulin (red) or glucagon (green). Scale bars = 25 µM |
Knockdown of zranb3 reduces β-cell number in zebrafish larvae. Injection of antisense oligonucleotide morpholinos (MO) targeting zranb3 reduced β-cells in 5 dpf larval zebrafish, detected by Tg(ins:mCherry) expression imaged via whole-mount confocal microscopy (a, b). c Depth coding of confocal microscopy images of whole-mount islets reveals individual cell resolution of β-cells in 5 dpf larval islets, color coded by depth (distal to proximal) along the z-axis. d, eWhole-mount immunostaining of MO-injected Tg(ins:mCherry) larval islets using antibody against glucagon (green). f Quantification of β-cell number in 5 dpf larval islets of Tg(ins:mCherry) injected with indicated concentrations of MO against zranb. (n = 17–19). Bars represent average across groups. Error bars represent SEM. p Values are from the t test. Scale bars = 25 µM |
ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions. PHENOTYPE:
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ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions. EXPRESSION / LABELING:
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(A) Validation of the efficacy of the zranb3 MO (5ng) to significantly suppress zranb3 mRNA expression without inducing off-target toxicity evaluated using the presence of a marker of MO-induced toxicity, the delta113 isoform of p53. (B) Wholemount bright field images of control MO and zranb3 MO-injected larvae shows no morphological developmental defects at 5 dpf PHENOTYPE:
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Glucose uptake as determined by retinal fluorescence after 2-NBDG treatment. (A) Quantification of glucose uptake at various concentrations in wild-type and zranb3 MO-injected larvae by quantification of 2-NBDG fluorescence. (B) Fluorescent 2-NBDG staining in control (left panel) and zranb3 MO-injected (right panel) larvae PHENOTYPE:
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ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions. PHENOTYPE:
|