Establishment and identification of a mouse model of vascular endothelial cell knockout DEPTOR gene

doi:10.3969/j.issn.2095-4344.1316

Abstract

Abstract:

BACKGROUND: There are few studies on DEPTOR and vascular diseases, and no studies have been found in animals. Therefore, the establishment of a new mouse model of vascular endothelial specific knockout DEPTOR is important for studying the relationship between DEPTOR and vascular diseases.
OBJECTIVE: To establish and identify a mouse model of vascular endothelial cell knockout DEPTOR gene.
METHODS: Cre mice and DEPTORflox/+ mice were purchased from the Jackson Laboratory, and C57 mice were provided by Huazhong University of Science and Technology. The study was approved by the Animal Ethic Committee of General Hospital of Middle Theater Command of Chinese PLA, approval number: 20120034. Five DEPTORflox/+male mice were selected to mate with 10 DEPTORflox/+ female mice, and 35 F1 progeny mice with genotype of EPTORflox/flox and DEPTORflox/+ were obtained and mated with 8 vascular endothelial cells specifically expressing Tek recombinase Cre mice. Finally 65 mice with genotypes of Tek-Cre+ x DEPTORflox/flox and DEPTORflox/flox progeny were obtained. The DEPTORflox and Cre genotypes were identified by PCR, and the body length and body mass of Tek-Cre+ x DEPTORflox/flox mice and DEPTORflox/flox mice at 2 months were recorded. The expression of DEPTOR protein in the mouse liver tissues was detected by western blot assay, and immunofluorescence was used to detect the expression of DEPTOR in the mouse vascular endothelial cells.
RESULTS AND CONCLUSION: (1) There were 25 Tek-Cre+ x DEPTORflox/flox mice and 40 DEPTORflox/flox mice, with the body length of (18.61±1.14) and (18.65±1.40) cm, respectively, and body mass of (25.84±1.99) and (25.06±2.15) g, respectively (both P > 0.05). (2) The relative expression level of DEPTOR protein in the Tek-Cre+ x DEPTORflox/flox mice and DEPTORflox/flox mice was 0.28±0.02 and 0.82±0.04, respectively (P < 0.05). (3) The number of DEPTOR-positive cell in vascular endothelial cells was 73.67±2.87 and 10.33±1.54, respectively (P < 0.05). (4) The results indicate that DEPTOR gene is successfully knocked out in Tek-Cre+ x DEPTORflox/flox mice. The homozygous mouse model of vascular endothelial cell knockout DEPTOR gene is successfully constructed and identified by genotype and protein tissue level.

Key words: vascular endothelial cells, DEPTOR mice, Cre/loxp technology, DEPTOR gene, DEPTOR homozygote, gene knockout

CLC Number:

Ding Yan, Meng Biying, Xiang Guangda. Establishment and identification of a mouse model of vascular endothelial cell knockout DEPTOR gene[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(23): 3698-3704.

Figures/Tables 4

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