FIGURE

Fig. 4

ID

ZDB-FIG-091016-29

Publication

Anderson et al., 2009 - Loss of Dnmt1 catalytic activity reveals multiple roles for DNA methylation during pancreas development and regeneration

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Fig. 4

Acinar cell death in dnmt1 mutants occurs in part by p53-dependent apoptosis. (A, B) DRAQ5 staining of DNA in 84 hpf WT (A) and dnmt1^s872 mutant (B) larvae. Pyknotic nuclei (arrows) are frequently observed in the pancreas of dnmt1 mutants, but not WT. (C, D) Apoptosis detection by TUNEL labeling of 84 hpf WT (C) and dnmt1 mutant (D) larvae in Tg(ptf1a:EGFP)^jh1; Tg(ins:dsRed)^m1081 background. C′/C″ and D′/D″ are transverse sections of similar samples at the approximate planes indicated in panels C and D, respectively. (E) Quantification of TUNEL⁺ acinar cells. (F–I) Expression of p53 mRNA in WT (F, H) and dnmt1^s872 mutants (G, I); expression appears dramatically increased in mutants. (J) Fold-change of p53 and target genes mdm2 and p21/waf1 in dnmt1^s872 mutants vs. WT by real-time RT-PCR. (K–N) 100 hpf WT (K, L) and dnmt1^s872 mutant (M, N) larvae, which are either uninjected controls (K, M) or p53 morpholino-injected (p53MO; L, N). p53MO-injected dnmt1^s872 mutants show increased persistence of acinar tissue. (O) More than 82% of p53MO-injected dnmt1^s872 mutants showed rescue (green); n = 22 (control), n = 45 (p53MO). Error bars = SEM. Significance assessed by Student′s t-test.

Expression Data

Genes:	DsRed EGFP tp53
Fish:	WT dnmt1^s872/s872 jh1Tg; m1018Tg dnmt1^s872/s872; jh1Tg; m1018Tg jh1Tg; m1018Tg + MO4-tp53 dnmt1^s872/s872; jh1Tg; m1018Tg + MO4-tp53
Knockdown Reagent:	MO4-tp53
Anatomical Terms:	endocrine pancreas exocrine pancreas head intestinal bulb liver midbrain hindbrain boundary pancreas pharyngeal arch
Stage Range:	Protruding-mouth to Day 4

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	dnmt1^s872/s872 dnmt1^s872/s872; jh1Tg; m1018Tg dnmt1^s872/s872; jh1Tg; m1018Tg + MO4-tp53
Knockdown Reagent:	MO4-tp53
Observed In:	apoptotic process exocrine pancreas exocrine pancreas nucleus
Stage Range:	Protruding-mouth to Day 4

Phenotype Detail

Acknowledgments

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Reprinted from Developmental Biology, 334(1), Anderson, R.M., Bosch, J.A., Goll, M.G., Hesselson, D., Dong, P.D., Shin, D., Chi, N.C., Shin, C.H., Schlegel, A., Halpern, M., and Stainier, D.Y., Loss of Dnmt1 catalytic activity reveals multiple roles for DNA methylation during pancreas development and regeneration, 213-223, Copyright (2009) with permission from Elsevier. Full text @ Dev. Biol.