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
|
