tff2-knockout mice  (International Mouse Phenotyping Consortium)

Journal: Diabetes

Article Title: Trefoil Factor 2 Expressed by the Murine Pancreatic Acinar Cells Is Required for the Development of Islets and for β-Cell Function During Aging

doi: 10.2337/db23-0490

Figure Lengend Snippet: Loss of Tff2 in the E16.5 pancreas leads to a reduction of islet and β-cells. A : Volcano plot showing a total of 1,143 DEGs in mutant compared with control pancreata at E16.5, with 448 downregulated and 695 upregulated DEGs ( n = 3 pancreata per group). FC, fold change; FDR, false discovery rate. B : Hallmark pathways from GSEA. C : List of the top 15 downregulated DEGs. D : Reads per kilobase of transcript per million reads mapped (RPKM) levels of canonical pancreatic β-cell genes. E : Conventional qRT-PCR analysis of selected endocrine cell genes relative to β-actin ( n = 4–6 pancreata). Error bars represent SEM. F and G : Representative co-IF images from control ( Tff2 fl/fl ) and mutant ( Pdx1:Cre;Tff2 fl/fl ) embryos (E16.5) double-stained for Chromogranin A (ChgA: red) and DBA (green) ( F ) or Tff2 (red) and Insulin (green) ( G ) and DAPI (blue). Scale bar = 50 μm. H–K : Quantification of total islet, β, and ductal mass ratios relative to total pancreas area (10–16 sections 100 μm apart per pancreas, n = 4 pancreata). The ratio of total Chromogranin A + area (square micrometers) divided by total pancreas area (square micrometers) ( H ), the ratio of total insulin + area (square micrometers) divided by total pancreas area (square micrometers) ( I ), total pancreas area (square micrometers) ( J ), and the ratio of total DBA + area (square micrometers) divided by total pancreas area (square micrometers) ( K ) ( n = 4 pancreata). Error bars represent SD. * P < 0.05, ** P < 0.01 and not significant (ns) determined by two-tailed Student t test for E and H – K and unpaired t test with Holm-Šídák method for D .

Article Snippet: The mice harboring conditional knockout allele for Tff2 were designed and produced by Ozgene Pty Ltd. (Perth, Australia) ( Supplementary Fig. 2 ).

Techniques: Mutagenesis, Control, Quantitative RT-PCR, Staining, Two Tailed Test

Journal: Diabetes

Article Title: Trefoil Factor 2 Expressed by the Murine Pancreatic Acinar Cells Is Required for the Development of Islets and for β-Cell Function During Aging

doi: 10.2337/db23-0490

Figure Lengend Snippet: Upon reaching 1 year of age, homozygous mutant mice exhibit higher body weights and fasting blood glucose levels as well as IGT. A and B : The body weight of control ( Tff2 fl/fl ; black), heterozygous ( Pdx1:Cre;Tff2 fl/+ ; blue), and homozygous ( Pdx1:Cre;Tff2 fl/fl ; red) male ( A ) and female ( B ) mice was monitored from 10 to 60 weeks ( n = 5–41 for males, n = 5–40 for females). C and D : Blood glucose levels after a 6-h fasting period were monitored from 10 to 60 weeks of age for male ( C ) and female ( D ) cohorts ( n = 5–41 for males, n = 3–40 for females). E–J : IP-GTT in male and female cohorts at 20 ( E and F ), 40 ( G and H ), and 60 weeks ( I and J ) ( n = 5–41 for males, n = 5–40 for females). Right graphs in E – J depict quantification of the AUC from 0 to 90 min after intraperitoneal injection of d -glucose (2 g/kg body weight). Error bars represent SD. * P < 0.05, ** P < 0.01, *** P < 0.001 and not significant (ns) determined by one-way ANOVA with Tukey post hoc test ( A – J ).

Article Snippet: The mice harboring conditional knockout allele for Tff2 were designed and produced by Ozgene Pty Ltd. (Perth, Australia) ( Supplementary Fig. 2 ).

Techniques: Mutagenesis, Control, Injection

Journal: Diabetes

Article Title: Trefoil Factor 2 Expressed by the Murine Pancreatic Acinar Cells Is Required for the Development of Islets and for β-Cell Function During Aging

doi: 10.2337/db23-0490

Figure Lengend Snippet: Aged homozygous mutants exhibit smaller pancreas organ size and impaired second-phase insulin secretion. A : Representative brightfield images of 1-year-old (1 yr) control ( Tff2 fl/fl ) and homozygous mutant ( Pdx1:Cre;Tff2 fl/fl ) pancreata. Scale bar = 1 mm. B : Body weight, pancreas weight, and pancreas-to-body weight ratio for aged males ( n = 14–22 for males). Error bars represent SD. C : Representative photomicrographs of hematoxylin-eosin–stained aged control and mutant pancreata. Scale bar = 1 mm. D–F : Quantification of total pancreas area (square micrometers) ( D ), exocrine cell mass ( E ), and islet cell mass ( F ) (five to eight sections 100 μm apart for each pancreas, n = 7–8 male mice). Error bars represent SD. G : Representative photomicrographs of immunohistochemistry staining of insulin (red) and hematoxylin (blue) in aged control and mutant pancreata. Scale bar = 1 mm. H–J : Quantification of total pancreas area (square micrometers) ( H ), the ratio of total insulin + area (square micrometers) divided by total pancreas area ( I ), and total β-cell mass ( J ) relative to total pancreas weight (five to eight sections 100 μm apart for each pancreas, n = 7 male mice). Error bars represent SD. K and L : In vitro GSIS in static incubation for cultured islets isolated from aged control and homozygous mutant male mice ( n = 4–7). Error bars represent SEM. M : Protein content ( N ) of islets from M ( n = 3–4 islet batches from four to seven male mice). Error bars represent SEM. N : Islets were isolated from aged male control or mutant mice and handpicked into groups of 40 and layered onto bead columns for parallel perifusion analysis. The islets were preincubated for 30 min in low glucose (2.8 mmol/L) followed by basal sample collection (1–13 min) to establish a baseline. Glucose was then elevated to 20 mmol/L for 35 min. Eluted fractions were collected at 1-min intervals at a flow rate of 0.3 mL/min, and secreted insulin was measured by radioimmunoassay. O : Quantification of the AUC for first-phase (14 to 20 min) and second-phase (22 to 45 min) insulin release from islets isolated from aged male control or mutant mice ( n = 4 islet batches from eight male mice). Error bars represent SD. P : Conventional qRT-PCR analyses of Tff2 and Tff3 relative to β-actin in exocrine tissue fraction after islet isolation shown in O ( n = 8 mice). Error bars represent SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 and not significant (ns) determined by two-tailed unpaired t test for H–J , O , and P or with Welch correction for B , D–F , L , and M .

Article Snippet: The mice harboring conditional knockout allele for Tff2 were designed and produced by Ozgene Pty Ltd. (Perth, Australia) ( Supplementary Fig. 2 ).

Techniques: Control, Mutagenesis, Staining, Immunohistochemistry, In Vitro, Incubation, Cell Culture, Isolation, RIA Assay, Quantitative RT-PCR, Two Tailed Test

Journal: Diabetes

Article Title: Trefoil Factor 2 Expressed by the Murine Pancreatic Acinar Cells Is Required for the Development of Islets and for β-Cell Function During Aging

doi: 10.2337/db23-0490

Figure Lengend Snippet: Tff2 protein is preferentially expressed by wild-type pancreatic acinar cells. A : Co-IF staining of Tff2 (red), insulin (green), and DAPI (blue) at E16.5, E18.5, P0, P7, and 6 weeks old (6 wks). Control tissue (6 wks, bottom right) treated with secondary antibodies only was used as negative control to adjust signal levels for Tff2. B : Co-IF staining of Tff2 (red), amylase (green), and DAPI (blue) at E16.5, P7, and 6 wks. White arrows indicate Tff2 + amylase + cells (yellow). C : Co-IF staining of Tff2 (red), Cpa-1 (green), and DAPI (blue) at P7 and 6 wks. D : Co-IF staining of Tff2 (red), DBA (green), and DAPI (blue) at P7 and 6 wks. E : Co-IF staining of Tff2 (red), EpCAM (green), and DAPI (blue) in P7 positive control tissues (pancreas, duodenum, stomach). Right panels show a × 4.9 magnification for A to E or × 13 magnification for B E16.5 or × 1.1 magnification for B P7 of the area marked by a white square. Yellow dotted lines and asterisk (*) outline islet/β-cell areas. Blue dotted lines outline ductal cell areas. Scale bar = 50 μm.

Article Snippet: The mice harboring conditional knockout allele for Tff2 were designed and produced by Ozgene Pty Ltd. (Perth, Australia) ( Supplementary Fig. 2 ).

Techniques: Staining, Control, Negative Control, Positive Control