Establishment and phenotypic analysis of mesenchymal cell bicaudal-C gene conditional knockout mice models
10.3760/cma.j.cn114453-20230321-00058
- VernacularTitle:间质细胞双尾C基因条件性敲除小鼠模型建立及其表型分析
- Author:
Yanshuang YIN
1
;
Xiu LI
;
Zhigang YANG
;
Shize MA
;
Yan CAO
;
Ran XIAO
Author Information
1. 中国医学科学院北京协和医学院整形外科医院研究中心 中国医学科学院体表组织器官再造研究重点实验室，北京 100144
- Keywords:
Mice, knockout;
Stromal cells;
Bicaudal-C gene;
Conditional gene knockout;
Tamoxifen
- From:
Chinese Journal of Plastic Surgery
2023;39(9):1003-1009
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish mesenchymal cell bicaudal-C (Bicc1) gene conditional knockout mice models and analyze their phenotypes.Methods:Bicc1 f/+ mice were crossed with Pdgfra promoter-driven Cre mice to obtain the offspring mice. Genomic DNA was extracted from the toe and tail tissues from 1-2 weeks old mice, amplified by PCR and detected at the DNA level by agarose gel electrophoresis. Three Bicc1 gene conditional knockout mice (experimental group) and three wild-type mice (control group) were selected after identification and grew to 3 weeks of age for follow-up experiments. The Bicc1 gene was knocked out by the induction of tamoxifen intraperitoneal injection. After 1 week, the kidney, skeletal muscle, skin and adipose tissue samples were collected. Real-time quantitative PCR (RT-qPCR) was performed to determine the expression levels of Bicc1 mRNA in the collected tissue samples. HE and Masson staining were performed with tissue samples fixed in 10% paraformaldehyde, and observed with a light microscope. The SPSS 28.0 software was used to analyze the data, t-test was used for comparison between groups, and P<0.05 was considered statistically significant. Results:Mesenchymal cell Bicc1 gene conditional knockout mice models were obtained by breeding, and the genotype was Bicc1 f/fCre +/-. The genotype of the wild-type mice was Bicc1 f/fCre -/-. RT-qPCR showed that the expression levels of Bicc1 mRNA in kidney, skeletal muscle, skin and adipose tissue of the experimental mice were significantly lower than those of the control group (all P<0.01). HE staining and Masson staining showed that compared with the control group, glomerular atrophy could be observed in the experimental group, renal capsules were irregular in shape, and some renal capsules disappeared. The arrangement of skeletal muscle fibers were loose and scattered, and the accumulation of muscle fibers was not dense. There were no significant differences between the skin and adipose tissue. Conclusion:Mesenchymal cell Bicc1 gene conditional knockout mice models were successfully established, which could provide models for studying the mechanisms of action of Bicc1 gene in different tissues and organs. Mesenchymal cell conditional Bicc1 gene knockout affected the phenotypes of kidney and skeletal muscle in mice.
