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Establishment of the mouse model of conditional knockout of Apr3 and preliminary study on the phenotypes

TAN Dong-qiong, YE Jun, HAN Lian-shu, QIU Wen-juan, GU Xue-fan, ZHANG Hui-wen   

  1. Department of Pediatric Endocrinology and Genetic Metabolism, Shanghai Institute for  Pediatric Research, Xinhua Hospital,Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China
  • Online:2015-08-28 Published:2015-09-30
  • Supported by:

    National Nature Science Foundation of China, 81071121

Abstract:

Objective  To establish the mouse model of conditional knockout of apoptosis related protein 3 (Apr3) and preliminary explore the phenotypes. Methods  The knockout plan was designed and Apr3(-/+) chimeric mice were obtained and bred to get Apr3(-/-) mice. The Apr3(-/-) homozygous mice with complete knockout of Apr3 were obtained by inbreeding and wild type control mice. DNAs of genome of mouse tail tissues were extracted. Wild type and knockout gene fragments were amplified by PCR and the phenotypes of mice were determined at DNA level. RNAs and proteins of mouse livers were extracted and the expression of Apr3 gene was detected by RT-PCR and Western blotting. The genetic traits and trend of body weight change of Apr3 knockout mice and indexes that might affect the growth, development, and metabolism, such as blood glucose, blood lipid, and liver function, were observed. Major lysosomal accumulated organs were hematoxylin-eosin stained and observed. Results  The mouse model of Apr3 knockout was successfully established. Apr3(-/-) mice were survived and fertile, but the body weight of two-month-old Apr3(-/-) mice was smaller than that of wild type mice. Results of the liver function test showed that the aspartate aminotransferase (AST) level of two-month-old Apr3(-/-) mice was higher than that of wild type mice (P<0.05). Results of H-E staining indicated significant degeneration and edema of liver cells and periportal inflammatory cell infiltration. Conclusion  The mouse model of conditional knockout of Apr3 is established. The knockout of Apr3 causes low body weight, abnormal morphology of liver tissue, and elevated AST of two-month-old mice.

Key words: Apr3 gene, knockout, phenotype;mouse