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Science 2 November 2007:
Vol. 318. no. 5851, pp. 806 - 809
DOI: 10.1126/science.1146812
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Reports

Menin Controls Growth of Pancreatic ß-Cells in Pregnant Mice and Promotes Gestational Diabetes Mellitus

Satyajit K. Karnik1, Hainan Chen1*, Graeme W. McLean1*, Jeremy J. Heit1*, Xueying Gu1, Andrew Y. Zhang1, Magali Fontaine2, Michael H. Yen1,3 and Seung K. Kim1,3{dagger}

1 Department of Developmental Biology, Stanford University, Stanford, CA 94305, USA.
2 Department of Pathology, Stanford University, Stanford, CA 94305, USA.
3 Department of Medicine (Oncology Division), Stanford University, Stanford, CA 94305, USA.


Figure 1 Fig. 1. Dynamic regulation of menin and its target genes during facultative islet growth in pregnant mice. (A) Quantification of BrdU incorporation by maternal islet ß-cells (n = 3 to 6 mice sampled per time point). (B and C) Detection of insulin (green) and BrdU (red) in islets from nonpregnant (B) and 17-dpc pregnant mice (C). (D and E) Detection of insulin (green) and menin (red) in islets from nonpregnant (D) and 17-dpc pregnant female mice (E). Scale bar, 50 µm. (F and G) Real-time PCR analysis of Men1 and Mll (F) and Western blot analysis of the indicated proteins (G) from isolated maternal islets. (H) ChIP studies of menin and trimethylated H3K4 associated with p18 and p27 from nonpregnant, pregnant, and postpartum maternal islets (n = 3 to 6 mice). Data here and in Figs. 2, 3, 4 are presented as the means ± SD. *P < 0.05, **P < 0.01. Ab, antibody; IgG, immunoglobulin G; Menin, anti-menin Ab; Methyl, anti-H3K4 Ab; In, input DNA; IP, immunoprecipitate. [View Larger Version of this Image (47K GIF file)]
 

Figure 2 Fig. 2. Phenotypes from misexpression of menin in islets of pregnant mice. (A) Western blot analysis of the indicated proteins in islets from pregnant control and pregnant ßMen1 mice, both at 17 dpc, and from nonpregnant control mice. (B to E) Immunohistologic detection of nuclear menin [red, arrowheads in (D) and arrows in (E)] in islet ß-cells (green) from pregnant control (B and D) and pregnant ßMen1 (C and E) mice, both at 17 dpc. Scale bar, 50 µm (B and C) and 20 µm (D and E). (F) Blood glucose levels in ßMen1 mice (black) and controls (gray) fed ad libitum and exposed to Dox before, during, and after pregnancy (n = 10 to 15 mice per group). (G) Intraperitoneal glucose tolerance tests of Dox-exposed pregnant ßMen1 (black) and pregnant control mice (gray), both at 16 dpc (n = 3 to 6 mice per group). Calculated area under the curve was 25 ± 0.4 area units (ßMen1) versus 22 ± 0.7 area units (controls), P <0.005.(H) Serum insulin concentrations, (I) pancreatic ß-cell mass, and (J) BrdU incorporation studies in 16- to 17-dpc pregnant ßMen1 mice (black), pregnant controls (gray), and nonpregnant controls (white), all administered Dox (n = 3 to 6 mice per genotype). (K and L) Detection of BrdU (red, arrows) and insulin (green) in pregnant control (K) and pregnant ßMen1 mice (L) at 17 dpc. Scalebar, 50 µm. *P < 0.05, **P < 0.01, ***P < 0.001. [View Larger Version of this Image (43K GIF file)]
 

Figure 3 Fig. 3. Stat5 and Bcl6 regulation of menin in maternal islets. (A and B) Detection of Insulin (green) and Stat5 (red) in islets from nonpregnant (A) and pregnant (B) mice. Scale bar, 50 µm. (C) Western blot detection of Stat5, phospho-Stat5, and Bcl6 in extracts of maternal islets. (D) ChIP analysis of Bcl6 association with the Men1, p18, and p27 loci in islets isolated from nonpregnant, pregnant, postpartum day 4, and postpartum day 21 mice. Primer pair 2 (PP2) for Men1 has been described previously (7). (E and F) Real-time RT-PCR analysis of Men1, p18, and p27 (E) or Bcl6 mRNA levels (F) in islets exposed to prolactin (black), vehicle (white), or to both prolactin and progesterone (gray) (n = 3 independent experiments). Real-time RT-PCR analysis of Bcl6 (G) or Men1, p18, and p27 mRNA levels (H) in islets from prolactin-infused (black) and control mice (white). (I) Quantification of BrdU incorporation by islet ß-cells in prolactin-infused (black) or control mice (white) (n = 8 mice per treatment). *P < 0.05, **P < 0.01, ***P < 0.001. [View Larger Version of this Image (47K GIF file)]
 

Figure 4 Fig. 4. Dynamic regulation of menin and its target genes during facultative islet growth in obese Ay mice. (A) Blood glucose concentration and (B) body mass in 12-week-old control and Ay littermate mice fed ad libitum. (C) Real-time RT-PCR analysis of Men1, p18, and p27 mRNA levels in islets from control or Ay littermate mice. (D) Western blot analysis of the indicated proteins in islets from control and Ay littermate mice. (E and F) Detection of insulin (green) and menin (red) in islet ß-cells from control (E) and Ay littermate (F) mice. Scale bar, 50 µm. [View Larger Version of this Image (48K GIF file)]
 

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