Inducible β-cell miR-21 overexpression

Tg(CAG-Z-miR-21-EGFP) [31 (link)] mice (backcrossed on a C57BL6/J background for >10 generations) were crossed with Ins1tm1(CreERT2)Thor [32 (link)] mice to generate Tg(βmiR-21) mice. Eight-week Tg(βmiR-21) mice and littermate controls (Cre+ and Cre-) were treated with 1 mg/day x 6 days intraperitoneal (IP) tamoxifen. IP glucose tolerance tests (IPGTTs) were performed 21-days post tamoxifen injection after overnight fast, using 2 g/kg body weight of glucose [28 (link)]. Tail vein glucose was determined (AlphaTRAK glucometer; Abbott) at 0, 10, 20, 30, 60, 90, and 120-min post injection. Insulin sensitivity was measured with IP insulin tolerance testing (IPITT) after a 2-h fast, using IP injection of 0.75 U/kg bodyweight of regular humulin-R insulin (Eli Lilly) [28 (link)]. Tail vein glucose was measured at 0, 10, 20, 30, and 60min post injection. Islets were isolated using collagenase 28 days after the initial tamoxifen injection [33 (link)].

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Ibrahim S., Johnson M., Stephens C.H., Xu J., Moore R., Mariani A., Contreras C., Syed F., Mirmira R.G., Anderson R.M, & Sims E.K. (2021). β-Cell pre-mir-21 induces dysfunction and loss of cellular identity by targeting transforming growth factor beta 2 (Tgfb2) and Smad family member 2 (Smad2) mRNAs. Molecular Metabolism, 53, 101289.

Publication 2021

AlphaTRAK glucometer

Bodyweight Collagenase Glucose Glucose tolerance tests Humulin Insulin sensitivity Intraperitoneal injection Mice Regular insulin Tail Tamoxifen Tolerance Vein

Corresponding Organization : University of Chicago

Other organizations : Molecular Biology Consortium

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Variable analysis

independent variables

Genotype (Tg(βmiR-21) mice vs. littermate controls (Cre+ and Cre-))
Tamoxifen treatment (1 mg/day x 6 days, intraperitoneal)

dependent variables

Glucose tolerance (IPGTT)
Insulin sensitivity (IPITT)
Islet isolation

control variables

Age of mice (8-week-old)
Genetic background (C57BL6/J, backcrossed for >10 generations)
Fasting duration (overnight for IPGTT, 2-h for IPITT)

positive controls

Cre+ littermate controls

negative controls

Cre- littermate controls

Annotations

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