ZFIN ID: ZDB-FIG-151110-3
Kimmel et al., 2015 - Diabetic pdx1-mutant zebrafish show conserved responses to nutrient overload and anti-glycemic treatment. Scientific Reports   5:14241 Full text @ Sci. Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibody:
Fish:
Condition:
Anatomical Terms:
Stage Range: Protruding-mouth to Days 7-13
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage Range: Protruding-mouth to Days 7-13

Fig. 3 Islet formation but impaired expansion in pdx1 mutants.

(a) Immunostaining for insulin (Ins, magenta) and GFP protein (green) in wild type and mutant NeuroD:eGFP+ embryos at 3 dpf. Maximum intensity projection (MIP) of confocal stacks. Scale bar, 20┬Ám. (b) Number of Ins-expressing cells from embryos as shown in (a). (wt, n = 9; mu, n = 10) **p < 0.001 (t-test). The line indicates the median, boxes span the 25th to 75th percentile, whiskers the 10th to 90th percentile. (dpf, days post fertilization) (c, top) Schematic of experiment to evaluate feeding induction of secondary islets. Embryos were provided with nutrient powder (SDS100) from day 5 until day 7, and imaged live on day 8 to detect secondary islet cells. (c, bottom) NeuroD:eGFP+ endocrine cells are detected outside of the islet in control pdx1+/+ larvae (left), which are absent in pdx1-/- mutants (right). The pancreas is outlined (gray) based on a simultaneously acquired brightfield image. (d) Quantitation of secondary islets formed in wild type (n = 11) and mutant (n = 11) larvae treated as in (c), ***p < 0.0001, t-test. (e) Representative 7 dpf NeuroD:eGFP+ larvae treated with Ly411575 starting on day 4 and examined live by fluorescence microscopy. Heterozygous and control larvae, but not mutants, show robust induction of new GFP+ endocrine cells in the pancreatic tail (arrows). (f) Quantitation of second wave islet cells from NeuroD:eGFP+ larvae treated as in (e). ***p < 0.0001; **p < 0.001; n.s., not significant (one-way ANOVA, Tukey Multiple Comparison test). n(pdx1+/+) = 8, n(pdx1+/-) = 13, n(pdx1-/-) = 7. In (c) and (e), the fluorescent signal in the prinicipal islet has been overexposed to reveal single cells and clusters in the surrounding pancreas. Single cells and clusters were quantitated as secondary islets.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
EGFP nl1Tg standard conditions Protruding-mouth endocrine pancreas IHC
Days 7-13 primary islet IFL
Days 7-13 secondary islet IFL
nl1Tg chemical treatment: LY-411575 Days 7-13 primary islet IFL
Days 7-13 secondary islet IFL
pdx1sa280/+ ; nl1Tg chemical treatment: LY-411575 Days 7-13 primary islet IFL
Days 7-13 secondary islet IFL
pdx1sa280/sa280 ; nl1Tg standard conditions Protruding-mouth endocrine pancreas IHC
Days 7-13 primary islet IFL
Days 7-13 Not Detected secondary islet IFL
pdx1sa280/sa280 ; nl1Tg chemical treatment: LY-411575 Days 7-13 primary islet IFL
Days 7-13 Not Detected secondary islet IFL
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-ins IHC nl1Tg standard conditions Protruding-mouth pancreatic B cell
IHC pdx1sa280/sa280 ; nl1Tg standard conditions Protruding-mouth pancreatic B cell
Phenotype Details
Fish Conditions Stage Phenotype
pdx1sa280/sa280 ; nl1Tg standard conditions Protruding-mouth endocrine pancreas decreased size, abnormal
Protruding-mouth pancreatic B cell decreased amount, abnormal
Days 7-13 secondary islet absent, abnormal
Days 7-13 secondary islet EGFP expression absent, abnormal
Days 7-13 secondary islet decreased amount, abnormal
pdx1sa280/sa280 ; nl1Tg chemical treatment: LY-411575 Days 7-13 secondary islet absent, abnormal
Days 7-13 secondary islet EGFP expression absent, abnormal
Days 7-13 secondary islet decreased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Scientific Reports for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Sci. Rep.