Stetsyuk et al., 2007 - Calsenilin is required for endocrine pancreas development in zebrafish. Developmental dynamics : an official publication of the American Association of Anatomists   236(6):1517-1525 Full text @ Dev. Dyn.

Fig. 1 Expression of calsenilin in the zebrafish pancreas. A: Search for new genes expressed in the pancreas: zebrafish at 24 hours postfertilization (hpf) were cohybridized with digoxigenin-labeled antisense probes (in blue) corresponding to the gene to be tested and fluorescein-labeled antisense insulin probe (in red). Calsenilin that was coexpressed with insulin was kept for further analysis. B: Expression of calsenilin during zebrafish development. Whole-mount in situ hybridization was performed with digoxigenin-labeled antisense calsenilin and insulin probes at different stages of development (21-somites, 24 hpf, 29 hpf, 32 hpf, 39 hpf). Arrows point to insulin-positive regions.

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Stage Range: 20-25 somites to Prim-25

Fig. 2 Coexpression of calsenilin and pancreatic hormones. Double fluorescent in situ hybridization in zebrafish embryos. Zebrafish at 24 hours postfertilization (hpf) and 30 hpf were cohybridized with digoxigenin-labeled antisense calsenilin probe in combination with either DNP-labeled antisense insulin, glucagon, or somatostatin2 probes. Calsenilin was revealed with tyramide-Cy3 substrate (in red) while insulin, glucagon, and somatostatin2 were revealed with tyramide-fluorescein isothiocyanate (in green). Images on the right column represent superposition of the respective left and middle confocal panels.

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Stage Range: Prim-5 to Prim-15

Fig. 3 Expression of calsenilin in mutants with affected retinoic acid synthesizing aldehyde dehydrogenase (RALDH; nls) and Notch (mib) signaling. Whole-mount in situ hybridization with digoxigenin-labeled antisense insulin and calsenilin probes on wild-type (WT), nls mutant defective in retinoic acid (RA) synthesis and mib mutants defective in the Delta-Notch pathway. Black and white arrows indicate insulin and calsenilin expression in the pancreas, respectively.

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Stage: Prim-5

Fig. 4 Pancreatic endocrine development is affected in calsenilin knockdown embryos. A: In vitro efficacy of morpholino antisense oligonucleotides directed against calsenilin. B: Whole-mount in situ hybridization on embryos injected with MO-calsenilin, MO3-calsenilin or with 5-misMO-calsenilin control antisense oligonucleotides. Zebrafish embryos at 24 hours postfertilization (hpf) or 40 hpf were hybridized with a digoxigenin-labeled antisense insulin probe. Quantification of insulin-positive cells either in 5-misMO-calsenilin, MO-calsenilin, or MO3-calsenilin morphants at stage 24 hpf. In knockdown embryos, the number of insulin-expressing cells was strongly decreased. ***P < 0.0001 C: Whole-mount in situ hybridization with digoxigenin-labeled antisense sst2, pdx1, isl1, and pax6.2 in MO-calsenilin and control 5-misMO-calsenilin embryos at 24 hpf. Note that in MO-calsenilin morphants, pancreatic endocrine cells were dispersed and did not associate to form an islet. Quantification of sst2-, isl1-, and Pax6.2-positive cells in wild-type or MO-calsenilin morphants at stage 24 hpf. *P < 0.01; ***P < 0.0001

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Stage Range: Prim-5 to Prim-25
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Stage Range: Prim-5 to Prim-25
Acknowledgments:
ZFIN wishes to thank the journal Developmental dynamics : an official publication of the American Association of Anatomists for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Dev. Dyn.