• GIPR, gastric Inhibitory polypeptide receptor, belongs to the G-protein coupled receptor family. GIPR is expressed in many tissues including pancreas, stomach, brain, liver, etc. This protein plays a crucial role in regulating insulin secretion, glucose, and lipid metabolism. Mutations or changes within this gene have been associated with various health conditions including obesity and diabetes.
• GLP1R, glucagon-like peptide-1 receptor, G protein-coupled receptor, 7 transmembrane proteins, human-mouse homology 91.79%. Widely distributed in multiple organs or tissues of the body, including the central nervous system, gastrointestinal tract, cardiovascular system, pancreas, liver, adipose tissue, muscle, etc.
• Glucagon acts through a 7-transmembrane class II receptor coupled to G proteins of either Gsα type or Gq type. Receptor activation of Gsα activates adenylate cyclase, resulting in cyclic adenosine monophosphate (cAMP) accumulation and increased protein kinase A activity and activation of the Gq receptor leads to an increase in intracellular Ca2+ through activation of phospholipase C.
• B-hGCGR/hGIPR/hGLP1R mice were obtained by mating B-hGCGR mice (110105), B-hGIPR mice (112714), and B-hGLP1R mice (170164).
• Mouse Gcgr mRNA was detectable in wild-type C57BL/6JNifdc mice but not in homozygous B-hGCGR/hGIPR/hGLP1R mice. Human GCGR mRNA was detectable only in homozygous B-hGCGR/hGIPR/hGLP1R mice, but not in wild-type mice.
• Mouse Gipr mRNA was detectable in wild-type C57BL/6 mice but not in homozygous B-hGCGR/hGIPR/hGLP1R mice. Human GIPR mRNA was detectable only in homozygous B-hGCGR/hGIPR/hGLP1R mice, but not in wild-type mice. GIPR was detected in liver, brain, muscle, adipose, and pancreas in wild-type mice and homozygous B-hGCGR/hGIPR/hGLP1R mice, as the antibody cross-recognizes both human and mouse GIPR.
• Human GLP1R mRNA was detectable only in homozygous B-hGCGR/hGIPR/hGLP1R mice, but not in wild-type mice. The pancreas, brain, lung, kidney, stomach, and large intestine show human GLP1R positive, liver and adipose show human GLP1R negative in homozygous B-hGCGR/hGIPR/hGLP1R mice. The pancreas, brain, lung, kidney, stomach, large intestine, small intestine, liver, and adipose tissue show mouse GLP1R negative in homozygous B-hGCGR/hGIPR/hGLP1R mice

B-hGCGR/hGIPR/hGLP1R mice were obtained by mating B-hGCGR mice (110105), B-hGIPR mice (112714), and B-hGLP1R mice (170164). For validation data of this mouse model, you can refer to the validation data from the related gene humanized mouse models.

• Gene targeting strategy for B-hGCGR mice. The exons 2-14 of the mouse Gcgr gene that encode the full-length protein were replaced by human GCGR exons 2-14 in B-hGCGR mice. The human GCGR protein expression will be driven by the endogenous mouse Gcgr promoter, while mouse Gcgr gene transcription and translation will be disrupted.
• Gene targeting strategy for B-hGIPR mice. The full coding sequences (CDS) of the mouse Gipr gene was replaced by human GIPR CDS + 3’UTR in B-hGIPR mice. The human GIPR protein expression will be driven by the endogenous mouse Gipr promoter, while mouse Gipr gene transcription and translation will be disrupted.
• Gene targeting strategy for B-hGLP1R mice. The full coding sequences (CDS) of the human GLP1R gene was inserted into the mouse Glp1r gene exon 1 in B-hGLP1R mice. The human GLP1R protein expression will be driven by the endogenous mouse Glp1r promoter, while mouse Glp1r gene transcription and translation will be disrupted

Strain-specific analysis of GCGR mRNA expression in wild-type C57BL/6JNifdc mice and B-hGCGR/hGIPR/hGLP1R mice by RT-PCR. Kidney RNA was isolated from wildtype C57BL/6JNifdc mice (+/+) and homozygous B-hGCGR/hGIPR/hGLP1R mice (H/H, H/H, H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse and human GCGR primers. Mouse Gcgr mRNA was detectable in wild-type C57BL/6JNifdc mice but not in homozygous B-hGCGR/hGIPR/hGLP1R mice. Human GCGR mRNA was detectable only in homozygous B-hGCGR/hGIPR/hGLP1R mice, but not in wild-type mice.

Strain-specific analysis of GIPR mRNA expression in wild-type C57BL/6JNifdc mice and B-hGCGR/hGIPR/hGLP1R mice by RT-PCR. Brain RNA was isolated from wildtype C57BL/6JNifdc mice (+/+) and homozygous B-hGCGR/hGIPR/hGLP1R mice (H/H, H/H, H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human GIPR primers. Mouse Gipr mRNA was detectable in wild-type C57BL/6 mice but not in homozygous B-hGCGR/hGIPR/hGLP1R mice. Human GIPR mRNA was detectable only in homozygous B-hGCGR/hGIPR/hGLP1R mice, but not in wild-type mice.

Strain-specific analysis of GLP1R mRNA expression in wild-type C57BL/6JNifdc mice and B-hGCGR/hGIPR/hGLP1R mice by RT-PCR. Lung RNA was isolated from wildtype C57BL/6JNifdc mice (+/+) and homozygous B-hGCGR/hGIPR/hGLP1R mice (H/H, H/H, H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with human GLP1R primers. Human GLP1R mRNA was detectable only in homozygous B-hGCGR/hGIPR/hGLP1R mice, but not in wild-type mice.

Western blot analysis of GIPR protein expression in homozygous B-hGCGR/hGIPR/hGLP1R mice. Various tissue lysates were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hGCGR/hGIPR/hGLP1R mice (H/H, H/H, H/H) and then analyzed by western blot with anti-GIPR antibody (Abcam, ab136266). 40 μg total proteins were loaded for western blotting analysis. GIPR was detected in liver, brain, muscle, adipose, and pancreas in wild-type mice and homozygous B-hGCGR/hGIPR/hGLP1R mice, as the antibody cross-recognizes both human and mouse GIPR.

Human GLP1R expression in different tissues of B-hGCGR/hGIPR/hGLP1R mice by IHC. Tissues were stained with human-specific antibodies GLP1R (Abcam, ab254352). The pancreas, brain, lung, kidney, stomach, and large intestine show human GLP1R positive (A, B, C, E, F, G). Liver and adipose show human GLP1R negative (D, H). Original magnification ×200. Abbreviations: IHC, immunohistochemistry.

Mouse GLP1R expression in different tissues of B-hGCGR/hGIPR/hGLP1R mice by IHC. Tissues were stained with mouse-specific antibodies GLP1R (Abcam, ab218532). The pancreas, brain, lung, kidney, stomach, large intestine, small intestine, liver, and adipose tissue show mouse GLP1R negative. Original magnification ×200. Abbreviations: IHC, immunohistochemistry. Positive Control: wild-type pancreas.