Ample Pairs

We show that the ample degree of a stable theory with trivial forking is preserved when we consider the corresponding theory of belles paires, if it exists. This result also applies to the theory of $H$-structures of a trivial theory of rank $1$.Comment: Research partially supported by the program MTM2014-59178-P. The second author conducted research with support of the programme ANR-13-BS01-0006 Valcomo. The third author would like to thank the European Research Council grant 33882

Terminal differentiation of intestinal goblet cells is affected in <i>agr2</i> morphants.

<p>Significant increases in immature Alcian blue-stained goblet cell numbers were detected in 104- (C, <i>n</i> = 46) and 120-hpf (G, <i>n</i> = 23) <i>agr2</i> morphants compared to those in either 104- (A, <i>n</i> = 40) and 120-hpf (E, <i>n</i> = 30) wild type or in 104- (B, <i>n</i> = 41) and 120-hpf (F, <i>n</i> = 27) agr2–5 mmMO1 and 5 mmMO2-coinjected embryos. Inset shows a mature and an immature goblet cell. Comparison of both immature and mature goblet cell numbers among <i>agr2</i> morphants, wild type or agr2–5 mmMO1 and 5 mmMO2-coinjected embryos at 104 and 120 hpf are shown (D, H). A Student's <i>t</i>-test was conducted to compare immature goblet cell numbers in <i>agr2</i> morphants with those in wild type or agr2–5 mmMO1 and 5 mmMO2-coinjected embryos. *p<0.001. Scale bars represent 100 µm.</p

Transmission electron microscopy shows abnormal goblet cell structures in <i>agr2</i> morphants.

<p>Mid-intestinal images of 104- and 120-hpf wild type (A, D), agr2–5 mmMO1 and 5 mmMO2-coinjected (B, E), and <i>agr2</i> morphants (C, F) are shown. ER ultrastructure at higher magnification is shown in insets. Arrows indicate mature goblet cells and arrowheads denote immature goblet cells. Scale bars represent 2 µm.</p

Intestinal cell proliferation is not altered in <i>agr2</i> morphants.

<p>Images of p-Histone H3-stained cells in the mid-intestines and posterior intestines of 104-hpf wild type embryos (A), agr2–5 mmMO1 and 5 mmMO2-coinjected embryos (B) and <i>agr2</i> morphants (C) are shown. Arrows indicate p-Histone H3-stained cells. (D) Comparison of the percentages of p-Histone H3-stained M phase cells among wild type embryos, agr2–5 mmMO1 and 5 mmMO2-coinjected embryos, and <i>agr2</i> morphants is shown. Error bars indicate the standard error. Scale bars represent 100 µm.</p

Enlarged areas of mature Alcian-blue stained goblet cells are detected in <i>agr2</i>-overexpressed embryos.

<p>Substantially increased areas of mature Alcian blue-stained intestinal goblet cells in 42% 104-hpf <i>agr2</i>-overexpressed (C, <i>n</i> = 29) embryos compared to <i>lacZ</i>-overexpressed (B, <i>n</i> = 20) embryos and wild type (A, <i>n</i> = 18) embryos were observed. Comparison of the area of Alcian blue-stained goblet cells in wild type, <i>agr2</i>- and <i>lacZ</i>-overexpressed embryos is shown (D). Arrows indicate examples of Alcian-blue stained goblet cells with enlarged areas. Student's <i>t</i>-test was conducted and *p<0.001. Scale bars represent 100 µm.</p

<i>agr2</i> morpholino antisense oligomer knockdown analyses.

<p>(A) Phenotype comparison among wild type, agr2–5 mmMO1 and 5 mmMO2-coinjected, and agr2-MO1 and agr2-MO2-coinjected embryos at 24 and 104 hpf. (B) Whole-mount immunohistochemistry demonstrates that coinjection of agr2-MO1 and agr2-MO2 prevents the synthesis of Agr2 protein in intestinal goblet cells. Confocal images of either wild type, agr2–5 mmMO1 and 5 mmMO2-coinjected, or agr2-MO1 and agr2-MO2-coinjected 104 hpf embryos were recorded under transmitted mode (a, d, g) or using 494/517 nm excitation/emission wavelengths (b, e, h). Merged images are shown (c, f, i). (C) Green fluorescence was not detected in agr2-MO1, agr2-MO2 and <i>CMV</i>-<i>agr2</i>-<i>mo</i>-<i>GFP</i> coinjected (c) 30 hpf embryos, whereas bright green fluorescence was observed in <i>CMV</i>-<i>agr2</i>-<i>mo</i>-<i>GFP</i>-injected (a) and agr2–5 mmMO1, agr2–5 mmMO2 and <i>CMV</i>-<i>agr2</i>-<i>mo</i>-<i>GFP</i> coinjected (b) 30 hpf embryos. Scale bars represent 100 µm.</p

Zebrafish <i>agr2</i> is solely expressed in intestinal goblet cells.

<p>Fluorescent whole-mount double <i>in situ</i> hybridization was conducted on 104-hours post fertilization (hpf) embryos using <i>agr2</i> (green) and <i>glucagon</i> (red) as RNA probes. Confocal images were recorded using excitation/emission wavelengths of 494/517 nm for fluorescein (A) and 550/570 nm for cyanine 3 (B). Merged image is shown (C). Arrows indicate <i>agr2</i>-expressing goblet cells and arrowheads specify <i>glucagon</i>-expressing enteroendocrine cells. Scale bars represent 100 µm.</p