Wong MH , Rubinfeld B , Gordon JI .

DK30292 NIDDK NIH HHS , DK37960 NIDDK NIH HHS , T32 H1O7275 PHS HHS , R01 DK030292 NIDDK NIH HHS , R37 DK030292 NIDDK NIH HHS

	Figure 2. Evidence for expression of the Fabpl–ΔN89β-catenin transgene. (A) RT-PCR analysis of RNAs isolated from the jejunum and skeletal muscle of a 6-wk-old-chimeric-transgenic mouse (ΔN89β-catenin) and from the jejunum of a 6-wk-old chimera generated using nontransfected ES cells (nl chimera). The arrow points to the expected size of the product generated from ΔN89β-catenin mRNA. (B) Duplicate immunoblots are shown each containing total cellular proteins isolated from the jejunum of a 6-wk-old normal control B6-ROSA26↔ 129/Sv chimeric mouse and a 6-wk-old B6-ROSA26↔ 129/Sv(ΔN89β-catenin) chimeric-transgenic animal (both with 30–40% 129/Sv contribution based on coat color). The blot on the left of the panel was probed with antibodies that recognize the myc epitope tag present at the NH2 terminus of the 80-kD ΔN89β-catenin mutant (arrow). The blot on the right of the panel was probed with antibodies raised against the COOH terminus of β-catenin (molecular mass of wild-type β-catenin = 91 kD). (C) PLP-fixed frozen section of a polyclonal villus from a B6-ROSA26↔ 129/Sv(ΔN89β-catenin) mouse. The section was incubated with affinity-purified rabbit antibodies to the myc tag, Cy3-tagged donkey anti–rabbit Ig, and the nuclear stain bis-benzimide (dark blue). Myc-tagged ΔN89β-catenin (magenta) is prominently represented in 129/Sv but not B6-ROSA26 villus epithelial cells. (Genotyping was accomplished by staining an adjacent serial section with antibodies to β-gal.) (D) Section of a polyclonal villus from a normal chimeric mouse stained with the same reagents as in C. 129/Sv epithelial cells lack the transgene, and therefore do not contain any myc-tagged protein. Bars: (C and D) 25 μm.
	Figure 3. Villus branching in 129/Sv(ΔN89β-catenin) epithelium. (A) Wholemount preparation of the jejunum from a 6-mo-old chimeric-transgenic mouse. Arrow, branched 129/Sv villus. (B) Section from the same wholemount stained with nuclear fast red. Each limb of the branched villus is equivalent in height. There are no obvious morphologic abnormalities in the crypts associated with this villus (arrows). (C) A branched polyclonal villus. Open arrow, a stripe of β-gal–positive (blue) B6-ROSA26 epithelium in one of the branches. The other branch is wholly 129/Sv (closed arrow; white). Inset, section of the branched villus incubated with rabbit antibodies to β-gal and Cy3-conjugated donkey anti–rabbit Ig. The stripe of B6-ROSA26 epithelium in one of the branches appears red. Closed arrow, the other, β-gal–negative, 129/Sv branch. Bars: (B and C) 25 μm.
	Figure 4. Quantitation of villus branching. Branching was defined and quantitated as described in Materials and Methods. Branched villi were scored in the B6-ROSA26 and 129/Sv jejunal components of 6-mo-old normal chimeras (B6-ROSA26↔ 129/ Sv) and chimeric-transgenic (B6-ROSA26↔ 129/Sv[NΔ89β-catenin]) mice. Each line of chimeric-transgenic mice was derived from an independent clone of ES cells stably transfected with Fabpl–NΔ89β-catenin DNA. Mean values ±1 SD are plotted. Asterisk, the frequency of branched villi was <0.01%.
	Figure 5. Forced expression of ΔN89β-catenin has no discernible effects on epithelial cell differentiation. Frozen sections were prepared from PLP-fixed jejunums of 6-mo-old chimeric-transgenic animals. (A) A polyclonal villus stained with biotin-conjugated Dolichos biflorus agglutinin (DBA), Cy3-conjugated avidin, rabbit anti–β-gal, and FITC-conjugated donkey anti–rabbit Ig. Glycoconjugates containing GalNAcα3GalNAc and GalNAcα3Gal recognized by DBA appear yellow-orange; β-gal appears green. The polarity and differentiation of enterocytes appears to be unaffected, as judged by the distribution of these glycoconjugates in apical membranes and supranuclear Golgi apparatus (closed arrow). Similarly, based on their reaction with DBA, the number and differentiation of goblet cells (open arrows) is equivalent in the 129/Sv and B6-ROSA26 components of the polyclonal villus. (B) The base of a polyclonal villus with its crypt-villus junction indicated by closed arrows. Three crypts are seen (open arrows at their base): the one on the left is supplying cells to another villus. The section was incubated with rat anti-β4 integrin subunit, Cy3 donkey anti–rat Ig, rabbit anti–β-gal, FITC donkey anti–rabbit Ig, and bis-benzimide. Nuclei (blue); the β4 integrin subunit (orange); β-gal (green-brown). The location of β4 integrin at the base of epithelial cells and its distribution along the crypt-villus unit are unaffected by ΔN89β-catenin. (C) Villi sectioned perpendicular to their crypt-villus axis. The tight junction protein ZO-1 (orange) was detected with rat anti-ZO-1 and Cy3 donkey anti–rat Ig. β-Gal (green) was visualized with the same reagents used in the preceding sections. The levels and location of ZO-1 in the 129/Sv(ΔN89β-catenin) and B6-ROSA26 components of polyclonal villi are similar (e.g., open arrows). (D) Villi sectioned perpendicular to their crypt-villus axis as in C. The section was incubated with rat anti-β7 integrin, Cy3-donkey anti–rat Ig, rabbit anti-laminin, rabbit anti–β-gal, FITC donkey anti–rabbit Ig, and bis-benzimide. B6-ROSA26 cells exhibit diffuse staining of their cytoplasm due to the presence of β-gal (green). β7 integrin is confined to intraepithelial lymphocytes (orange). Comparable numbers of these cells are seen in the B6-ROSA26 and 129/ Sv(ΔN89β-catenin) components of polyclonal villi and in wholly 129/Sv(ΔN89β-catenin) villi. Laminin appears as linear green immunoreactivity underlying 129/Sv and B6-ROSA26 epithelium (e.g., closed arrows). The intensity of staining is similar under cells of both genotypes. Bars, 25 μm.
	Figure 6. ΔN89β-catenin stimulates proliferation and apoptosis in 129/Sv crypts. (A) The number of M-phase cells in juxtaposed B6-ROSA26 and 129/Sv jejunal crypt sections were scored in 6-mo-old normal chimeras, in two lines of chimeric-transgenic mice generated using separate ES cell clones stably transfected with Fabpl–ΔN89β-catenin DNA, and in two lines of chimeric-transgenic mice produced from independent ES cells clones stably transfected with Fabpl-mouse E-cadherin DNA (Hermiston et al., 1996). Each mouse served as its own control: the average number of M-phase cells in their 129/Sv crypts were divided by the average number of M-phase cells in their B6-ROSA26 crypts. Mean values ±1 SD obtained from animals in each group are plotted except for line 2 of B6↔ 129/Sv (E-cadherin) chimeras where only two animals were examined. Note that the stimulation of crypt proliferation is similar in mice belonging to a given line of B6-ROSA26↔ 129/Sv(ΔN89β-catenin) chimeras. Differences between lines produced from different Fabpl–ΔN89β-catenin ES cell clones were attributed to differences in the levels of expression of this transgene. (B) Apoptotic cells were scored in jejunal sections prepared from the same animals used to define the mitotic index in A.
	Figure 7. Forced expression of ΔN89β-catenin is associated with a slowing of epithelial migration. A 6-mo-old chimeric-transgenic mouse was pulse labeled with BrdU 60 h before killing. Left, a section stained with rabbit anti–β-gal and Cy3 donkey anti–rabbit Ig. The villus encompassed by the box is polyclonal. Right, an adjacent section of the polyclonal villus, stained with goat anti-BrdU, Cy3 donkey anti–goat Ig, and bis-benzimide. The leading edge of the column of BrdU–positive 129/Sv(ΔN89β-catenin) epithelial cells (magenta-colored nuclei) has only reached the middle portion of the villus (arrow), whereas the leading edge of the BrdU–positive B6-ROSA26 column is already near the villus tip. Bars, 25 μm.
	Figure 8. ΔN89β-Catenin expression results in augmented cellular pools of E-cadherin. Frozen sections were prepared from PLP-fixed jejunum recovered from a 6-mo-old B6-ROSA26↔ 129/ Sv(ΔN89β-catenin) chimeric-transgenic animal (A–D) and a 6-mo-old normal B6-ROSA26↔ 129/Sv chimera (E and F). (A) Polyclonal villus incubated with rabbit anti–β-catenin, Cy3 donkey anti–rabbit Ig, and bis-benzimide. Prominent β-catenin staining (red) is evident at the adherens junctions and basolateral surfaces of epithelial cells. No immunoreactive protein is detectable in 129/Sv or B6-ROSA26 nuclei. (B) Polyclonal villus incubated with rabbit anti-APC, Cy3 donkey anti–rabbit Ig, and bis-benzimide. The levels and intracellular distribution of APC (red) are similar in B6-ROSA26 and 129/Sv(ΔN89β-catenin) epithelial cells. (C and D) Villi from the chimeric-transgenic mouse that have been sectioned perpendicular their crypt-villus axis. (C) Section stained with rat anti–E-cadherin and Cy3-donkey anti– rat Ig. (D) Dual exposure of the same section after incubation with rabbit anti–β-gal and FITC donkey anti–rabbit Ig. Steady-state levels of E-cadherin are markedly increased in the 129/ Sv(ΔN89β-catenin) component of polyclonal villi. (E and F) Villi from a control normal chimera processed as in (C and D). E-Cadherin levels are comparable in the B6-ROSA26 and 129/Sv villus epithelium. Bars, 25 μm.

Aberle, Assembly of the cadherin-catenin complex in vitro with recombinant proteins. 1994, Pubmed

Aberle, Assembly of the cadherin-catenin complex in vitro with recombinant proteins. 1994, Pubmed
Aberle, beta-catenin is a target for the ubiquitin-proteasome pathway. 1997, Pubmed
Barth, NH2-terminal deletion of beta-catenin results in stable colocalization of mutant beta-catenin with adenomatous polyposis coli protein and altered MDCK cell adhesion. 1997, Pubmed
Behrens, Functional interaction of beta-catenin with the transcription factor LEF-1. 1996, Pubmed , Xenbase
Brown, A retrovirus vector expressing the putative mammary oncogene int-1 causes partial transformation of a mammary epithelial cell line. 1986, Pubmed
Brunner, pangolin encodes a Lef-1 homologue that acts downstream of Armadillo to transduce the Wingless signal in Drosophila. 1997, Pubmed
Cadigan, Wnt signaling: a common theme in animal development. 1997, Pubmed , Xenbase
Chandrasekaran, Use of normal and transgenic mice to examine the relationship between terminal differentiation of intestinal epithelial cells and accumulation of their cell cycle regulators. 1996, Pubmed
Cheng, Origin, differentiation and renewal of the four main epithelial cell types in the mouse small intestine. I. Columnar cell. 1974, Pubmed
Cheng, Origin, differentiation and renewal of the four main epithelial cell types in the mouse small intestine. II. Mucous cells. 1974, Pubmed
Cheng, Origin, differentiation and renewal of the four main epithelial cell types in the mouse small intestine. III. Entero-endocrine cells. 1974, Pubmed
Cheng, Origin, differentiation and renewal of the four main epithelial cell types in the mouse small intestine. IV. Paneth cells. 1974, Pubmed
Cheng, Origin, differentiation and renewal of the four main epithelial cell types in the mouse small intestine. V. Unitarian Theory of the origin of the four epithelial cell types. 1974, Pubmed
Cohn, Use of transgenic mice to map cis-acting elements in the intestinal fatty acid binding protein gene (Fabpi) that control its cell lineage-specific and regional patterns of expression along the duodenal-colonic and crypt-villus axes of the gut epithelium. 1992, Pubmed
Cook, Wingless inactivates glycogen synthase kinase-3 via an intracellular signalling pathway which involves a protein kinase C. 1996, Pubmed
Coopersmith, Bi-transgenic mice reveal that K-rasVal12 augments a p53-independent apoptosis when small intestinal villus enterocytes reenter the cell cycle. 1997, Pubmed
Cox, Armadillo is required for adherens junction assembly, cell polarity, and morphogenesis during Drosophila embryogenesis. 1996, Pubmed
Dickinson, Evidence for a mitogenic effect of Wnt-1 in the developing mammalian central nervous system. 1994, Pubmed
Fagotto, Binding to cadherins antagonizes the signaling activity of beta-catenin during axis formation in Xenopus. 1996, Pubmed , Xenbase
Falk, Lectins are sensitive tools for defining the differentiation programs of mouse gut epithelial cell lineages. 1994, Pubmed
Friedrich, Promoter traps in embryonic stem cells: a genetic screen to identify and mutate developmental genes in mice. 1991, Pubmed
Funayama, Embryonic axis induction by the armadillo repeat domain of beta-catenin: evidence for intracellular signaling. 1995, Pubmed , Xenbase
Garabedian, Examining the role of Paneth cells in the small intestine by lineage ablation in transgenic mice. 1997, Pubmed
Haegel, Lack of beta-catenin affects mouse development at gastrulation. 1995, Pubmed
Hall, Regulation of cell number in the mammalian gastrointestinal tract: the importance of apoptosis. 1994, Pubmed
Han, Gut reaction to Wnt signaling in worms. 1997, Pubmed
Heasman, Overexpression of cadherins and underexpression of beta-catenin inhibit dorsal mesoderm induction in early Xenopus embryos. 1994, Pubmed , Xenbase
Hermiston, Forced expression of E-cadherin in the mouse intestinal epithelium slows cell migration and provides evidence for nonautonomous regulation of cell fate in a self-renewing system. 1996, Pubmed
Hermiston, In vivo analysis of cadherin function in the mouse intestinal epithelium: essential roles in adhesion, maintenance of differentiation, and regulation of programmed cell death. 1995, Pubmed
Hermiston, Inflammatory bowel disease and adenomas in mice expressing a dominant negative N-cadherin. 1995, Pubmed
Hill, Protein determination using bicinchoninic acid in the presence of sulfhydryl reagents. 1988, Pubmed
Hinck, Dynamics of cadherin/catenin complex formation: novel protein interactions and pathways of complex assembly. 1994, Pubmed
Huber, Nuclear localization of beta-catenin by interaction with transcription factor LEF-1. 1996, Pubmed , Xenbase
Huber, Three-dimensional structure of the armadillo repeat region of beta-catenin. 1997, Pubmed
Ilyas, Beta-catenin mutations in cell lines established from human colorectal cancers. 1997, Pubmed
Jou, Genetic and biochemical dissection of protein linkages in the cadherin-catenin complex. 1995, Pubmed
Kim, Transgenic mouse models that explore the multistep hypothesis of intestinal neoplasia. 1993, Pubmed
Kinzler, Lessons from hereditary colorectal cancer. 1996, Pubmed
Korinek, Constitutive transcriptional activation by a beta-catenin-Tcf complex in APC-/- colon carcinoma. 1997, Pubmed
Laemmli, Cleavage of structural proteins during the assembly of the head of bacteriophage T4. 1970, Pubmed
Loeffler, Somatic mutation, monoclonality and stochastic models of stem cell organization in the intestinal crypt. 1993, Pubmed
McCrea, Induction of a secondary body axis in Xenopus by antibodies to beta-catenin. 1993, Pubmed , Xenbase
McLean, Periodate-lysine-paraformaldehyde fixative. A new fixation for immunoelectron microscopy. 1974, Pubmed
Miller, Signal transduction through beta-catenin and specification of cell fate during embryogenesis. 1996, Pubmed
Molenaar, XTcf-3 transcription factor mediates beta-catenin-induced axis formation in Xenopus embryos. 1996, Pubmed , Xenbase
Morin, Activation of beta-catenin-Tcf signaling in colon cancer by mutations in beta-catenin or APC. 1997, Pubmed
Munemitsu, Deletion of an amino-terminal sequence beta-catenin in vivo and promotes hyperphosporylation of the adenomatous polyposis coli tumor suppressor protein. 1996, Pubmed
Munemitsu, Regulation of intracellular beta-catenin levels by the adenomatous polyposis coli (APC) tumor-suppressor protein. 1995, Pubmed
Nagafuchi, Cell binding function of E-cadherin is regulated by the cytoplasmic domain. 1988, Pubmed
Noordermeer, dishevelled and armadillo act in the wingless signalling pathway in Drosophila. 1994, Pubmed
Nusse, A versatile transcriptional effector of Wingless signaling. 1997, Pubmed
Näthke, The adenomatous polyposis coli tumor suppressor protein localizes to plasma membrane sites involved in active cell migration. 1996, Pubmed
Orsulic, An in vivo structure-function study of armadillo, the beta-catenin homologue, reveals both separate and overlapping regions of the protein required for cell adhesion and for wingless signaling. 1996, Pubmed
Overduin, Solution structure of the epithelial cadherin domain responsible for selective cell adhesion. 1995, Pubmed
Ozawa, The cytoplasmic domain of the cell adhesion molecule uvomorulin associates with three independent proteins structurally related in different species. 1989, Pubmed
Ozawa, Uvomorulin-catenin complex formation is regulated by a specific domain in the cytoplasmic region of the cell adhesion molecule. 1990, Pubmed
Pai, Negative regulation of Armadillo, a Wingless effector in Drosophila. 1997, Pubmed
Pollack, Dynamics of beta-catenin interactions with APC protein regulate epithelial tubulogenesis. 1997, Pubmed
Potten, The intestinal epithelial stem cell: the mucosal governor. 1997, Pubmed
Riese, LEF-1, a nuclear factor coordinating signaling inputs from wingless and decapentaplegic. 1997, Pubmed
Rimm, Alpha 1(E)-catenin is an actin-binding and -bundling protein mediating the attachment of F-actin to the membrane adhesion complex. 1995, Pubmed
Rocheleau, Wnt signaling and an APC-related gene specify endoderm in early C. elegans embryos. 1997, Pubmed
Rubinfeld, Binding of GSK3beta to the APC-beta-catenin complex and regulation of complex assembly. 1996, Pubmed
Rubinfeld, Stabilization of beta-catenin by genetic defects in melanoma cell lines. 1997, Pubmed
Rubinfeld, Association of the APC gene product with beta-catenin. 1993, Pubmed
Schmidt, Cell migration pathway in the intestinal epithelium: an in situ marker system using mouse aggregation chimeras. 1985, Pubmed
Shapiro, Structural basis of cell-cell adhesion by cadherins. 1995, Pubmed
Shibata, Rapid colorectal adenoma formation initiated by conditional targeting of the Apc gene. 1997, Pubmed
Simon, Use of transgenic mice to map cis-acting elements in the liver fatty acid-binding protein gene (Fabpl) that regulate its cell lineage-specific, differentiation-dependent, and spatial patterns of expression in the gut epithelium and in the liver acinus. 1993, Pubmed
Su, Association of the APC tumor suppressor protein with catenins. 1993, Pubmed
Su, Multiple intestinal neoplasia caused by a mutation in the murine homolog of the APC gene. 1992, Pubmed
Sweetser, Mechanisms underlying generation of gradients in gene expression within the intestine: an analysis using transgenic mice containing fatty acid binding protein-human growth hormone fusion genes. 1988, Pubmed
Thorpe, Wnt signaling polarizes an early C. elegans blastomere to distinguish endoderm from mesoderm. 1997, Pubmed
Totafurno, Theoretical analysis of the flow of cells over villi of the small intestine. 1990, Pubmed
Totafurno, The crypt cycle. Crypt and villus production in the adult intestinal epithelium. 1987, Pubmed
Trahair, Use of transgenic mice to study the routing of secretory proteins in intestinal epithelial cells: analysis of human growth hormone compartmentalization as a function of cell type and differentiation. 1989, Pubmed
Tsukamoto, Expression of the int-1 gene in transgenic mice is associated with mammary gland hyperplasia and adenocarcinomas in male and female mice. 1988, Pubmed
Wong, Forced expression of the tumor suppressor adenomatosis polyposis coli protein induces disordered cell migration in the intestinal epithelium. 1996, Pubmed
Wright, The kinetics of villus cell populations in the mouse small intestine. I. Normal villi: the steady state requirement. 1982, Pubmed
Yanagawa, Accumulation of Armadillo induced by Wingless, Dishevelled, and dominant-negative Zeste-White 3 leads to elevated DE-cadherin in Drosophila clone 8 wing disc cells. 1997, Pubmed
Yost, The axis-inducing activity, stability, and subcellular distribution of beta-catenin is regulated in Xenopus embryos by glycogen synthase kinase 3. 1996, Pubmed , Xenbase
van de Wetering, Armadillo coactivates transcription driven by the product of the Drosophila segment polarity gene dTCF. 1997, Pubmed