请输入关键字
请输入关键字
订购
*国家
中国
美国
*省份
*城市
*姓名
*电话
*单位
*职位
*邮箱
*请输入验证码
*验证码
B-hGCGR mice
Strain Name C57BL/6-Gcgrtm1(GCGR)/Bcgen Common Name  B-hGCGR mice
Background C57BL/6 Catalog number  110105
Aliases 
GGR, GL-R,G, GR

Gene description


GCGR encodes a glucagon receptor that is important in controlling blood glucose levels. Defects in this gene are a cause of non-insulin-dependent diabetes mellitus (NIDDM). Activation of the human glucagon receptor (GCGR) by its endogenous ligand glucagon triggers the release of glucose from the liver during fasting, making it a potential drug target for type 2 diabetes.


mRNA expression analysis


from clipboard


Strain specific analysis of GCGR gene expression in WT and B-hGCGR mice by RT-PCR. Mouse GCGR mRNA was detectable only in liver cells of wild type C57BL/6 mice (+/+). Human GCGR mRNA was detectable only in homozygous B-hGCGR mice (H/H) , but not in wild type C57BL/6 mice (+/+). 


Protein expression analysis

from clipboard


Strain specific GCGR expression analysis in homozygous B-hGCGR mice by western blot. Kidney tissue was collected from wild type C57BL/6 mice (+/+) and homozygous B-hGCGR mice (H/H), and analyzed by western blot with anti-GCGR antibody. Mouse GCGR was detectable in wild type mice and homozygous B-hGCGR mice, as the antibody is crossly reactive with GCGR in human and mice. Human GCGR was exclusively detectable in homozygous B-hGCGR mice but not in wild type C57BL/6 mice.

Analysis of leukocytes cell subpopulation in B-hGCGR mice

from clipboard


Analysis of spleen leukocyte subpopulations by FACS
Splenocytes were isolated from female C57BL/6 and B-hGCGR mice (n=4, 7-week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hGCGR mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hGCGR in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in spleen. Values are expressed as mean ± SEM.

Analysis of T cell subpopulation in B-hGCGR mice


from clipboard


Analysis of spleen T cell subpopulations by FACS
Splenocytes were isolated from female C57BL/6 and B-hGCGR mice (n=4, 7-week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ T cells were gated for CD3+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of CD4+ T cells, CD8+ T cells and Tregs in homozygous B-hGCGR mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hGCGR in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell sub types in spleen. Values are expressed as mean ± SEM.

Analysis of leukocytes cell subpopulation in B-hGCGR mice
from clipboard
Analysis of subpopulation of leukocytes in lymph node by FACS
Lymph node was isolated from female C57BL/6 and B-hGCGR mice (n=4, 7-week-old). Flow cytometry analysis of the lymph node was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ T cells were used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells, and NK cells in homozygous B-hGCGR mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hGCGR in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in lymph node. Values are expressed as mean ± SEM.

Analysis of T cell subpopulation in B-hGCGR mice

from clipboard


Analysis of subpopulation of T cells in lymph node by FACS
Lymph node was isolated from female C57BL/6 and B-hGCGR mice (n=4, 7-week-old). Flow cytometry analysis of the lymph node was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ T cells were used for further analysis as indicated here. B. Results of FACS analysis. Percent of CD4+ T cells, CD8+ T cells and Tregs in homozygous B-hGCGR mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hGCGR in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in lymph node. Values are expressed as mean ± SEM.


Analysis of leukocytes cell subpopulation in B-hGCGR mice


from clipboard


Analysis of blood leukocyte subpopulations by FACS
Blood cells were isolated from female C57BL/6 and B-hGCGR mice (n=4, 7-week-old). Flow cytometry analysis of blood cells was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hGCGR mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hGCGR in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in blood. Values are expressed as mean ± SEM.

Analysis of T cell subpopulation in B-hGCGR mice

from clipboard


Analysis of subpopulation of T cells in blood by FACS
Blood cells were isolated from female C57BL/6 and B-hGCGR mice (n=4, 7-week-old). Flow cytometry analysis of blood cells was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ T cells were used for further analysis as indicated here. B. Results of FACS analysis. Percent of CD4+ T cells, CD8+ T cells and Tregs in homozygous B-hGCGR mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hGCGR in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in lymph node. Values are expressed as mean ± SEM.

In vivo efficacy of anti-human GCGR antibody with B-hGCGR mice

from clipboard


from clipboard


Experimental schedule for in vivo efficacy of anti-human GCGR antibody. Anti-human GCGR antibody-crotedumab (in house) was administered by intraperitoneal injection once a week on days 1 to 8. Blood were collected for analysis of blood glucose, insulin, glucagon and lipid on the days showed in the schematic diagram and the table. 


In vivo efficacy of anti-human GCGR antibody with B-hGCGR mice


from clipboard

Anti-human GCGR antibody reduces blood glucose in male B-hGCGR mice. 
A. Random blood glucose from male mice before and at multiple time points after injection of crotedumab (in house) or isotype control antibody (n = 6). B. Body weights. C. OGTT on day 4. D. Area under the curve for the OGTT shown in C. Serum levels of (E) insulin, (F) glucagon on Day 7. Anti-GCGR antibody reduced the random blood glucose, fasting blood glucose and OGTT compared to the isotype antibody in B-hGCGR mice. Serum levels of insulin was reduced slightly and glucagon level was significantly increased in the antibody treated group. All of the results in B-hGCGR mice were similar to those in the wild-type C57BL/6. 
Results indicated that the regulatory function on blood glucose in humanized B-hGCGR mice was similar to the wild-type C57BL/6. Anti-human GCGR antibody was efficacious in controlling blood glucose in B-hGCGR mice. Values are expressed as mean ± SEM. OGTT, oral glucose tolerance test. 


In vivo efficacy of anti-human GCGR antibody with B-hGCGR mice


from clipboard

Anti-human GCGR antibody improved lipid metabolism in male B-hGCGR mice. 

Wild-type C57BL/6 and B-hGCGR mice were treated with crotedumab (in house) or isotype control antibody (n = 6). Blood were collected on Day 11 for triglycerides and cholesterol analysis. Serum levels of TG (A) and TC (B). HDL-C (C) and LDL-C (D) levels in serum. Serum levels of TG was reduced, while TC, HDL-C and LDL-C were increased in the anti-human GCGR antibody treated male mice group compared to the isotype control. All of the results in B-hGCGR mice were similar to those in the wild-type C57BL/6. Results indicated that lipid metabolism in humanized B-hGCGR mice was similar to the wild-type C57BL/6. Results indicated that anti-human GCGR antibody was efficacious in controlling blood lipid in male B-hGCGR mice. Values are expressed as mean ± SEM. TG, triglycerides; TC, total cholesterol; HDL-C, high density lipoprotein cholesterol; LDL-C, low density lipoprotein cholesterol.