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First published online 20 August 2008
doi: 10.1242/dev.017137

Development 135, 3191-3196 (2008)
Published by The Company of Biologists 2008
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Conditional inactivation of Myc impairs development of the exocrine pancreas

Hassan Nakhai, Jens T. Siveke, Lidia Mendoza-Torres and Roland M. Schmid*

Department of Internal Medicine, Technical University of Munich, 81675 Munich, Germany.


Figure 1
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  		Fig. 1. Pancreata of PMyc+/- and PMycKO embryos at E18.5. (A,B) X-Gal staining of dissected intestinal tracts from heterozygous PMyc+/-;R26R and PMycKO;R26R embryos. (C,D) X-Gal stained sections of PMyc+/-;R26R and PMycKO;R26R pancreata. (E-I) Immunostaining for amylase (E,F, brown) and PTF1a (H,I, black, arrows) in PMyc+/-;R26R and PMycKO;R26R embryos. (G) Double-immunofluorescence for amylase (red) and β-galactosidase (green) in PMycKO;R26R embryo. Inserts in H,I represent a 2x enlargement. Scale bar: 50 µm. Abbreviations: p, pancreas; ac, acini.

 

Figure 2
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  		Fig. 2. Pancreatic buds of PMycKO embryos. (A-Q) Wild-type and PMycKO pancreatic buds were analyzed by immunohistochemistry. (A,B) Anti-Myc staining of dorsal buds (black, arrow) at E12.5. (C,D) Immunofluorescence staining of dorsal buds with anti-PDX1 antibody (green) at E13.5. (E,F) Double-immunofluorescence staining of dorsal buds with anti-glucagon (red) and anti-PDX1 (green) at E10.5. (G) Quantification of glucagon+ cell area in dorsal buds of wild-type and PMycKO at E10.5. (H,I) Immunostaining of dorsal buds with anti-neurogenin 3 at E12.5 (black arrows). (K,L) Anti-PHH3 staining of pancreatic buds (black, arrows) at E12.5. (N,O) Immunostaining of pancreatic buds with anti-cyclin D1 antibody (black, arrows) at E12.5. (P,Q) Immunostaining of dorsal buds with anti-CDK4 antibody (cytoplasmic, brown, arrows) at E13.5. (J,M) Quantification of the number of neurogenin 3+- and PHH3+ cells in buds of E12.5 wild-type and PMycKO embryos are shown. Each histogram represents the mean±s.d. for ventral and dorsal buds of three embryos each. Insets in A, P and Q represent a 2x enlargement and a 4x enlargement in B, N and O. Background in C and D was stained by DAPI. Abbreviations: l, liver; st, stomach; vb, ventral bud; db, dorsal bud. Scale bar: 50 µm.

 

Figure 3
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  		Fig. 3. Acinar cells in PMycKO embryos. Serial sections of E14.5 wild-type and PMycKO embryos stained for Myc (A,B, black, arrows) and amylase (C,D, brown, arrows). (E,F) Double-immunofluorescence staining of wild-type and PMycKO sections for amylase (green) and BrdU (red) at E15.5. Arrows indicate the BrdU-positive and arrowheads the BrdU-negative acinar cells. (G) BrdU+ nuclei were counted in amylase immunoreactive cells. Inserts in A and C represent a 4x enlargement and a 2x enlargement in E. Abbreviations: ac, acini; l, liver; p, pancreas. Scale bar: 50 µm.

 

Figure 4
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  		Fig. 4. Endocrine development in PMycKO pancreata. Immunofluorescence staining for insulin (A,B, arrows) in E13.5 dorsal pancreatic buds of wild-type and PMycKO sections. (C-J) Sections of E18.5 wild-type and PMycKO embryos were analyzed for expression of endocrine markers by immunostaining (brown, arrows). (K-P) X-gal stained sections of E18.5 Ptf1a-cre(ex1);R262R and PMycKO;R26R pancreata were immunostained for glucagon. (Q,R) The β and {alpha} cell masses were determined from anti-insulin- and anti-glucagon-stained multiple sections as described in the Materials and methods section. Inserts in I and J represent a 2x and a 4x enlargement respectively. Scale bar: 50 µm.

 

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© The Company of Biologists Ltd 2008