[Neural stem cell-specific peroxisome proliferator-activated receptor γ knockout mice: breeding and genetic...

To breed neual stem cell-specific peroxisome proliferator-activated receptor γ (PPARγ) knockout mice.

Two transgenic mouse models, namely B6.PPARγloxp/loxp and B6.Nestin-Cre were interbred, and the first- generation offsprings were backcrossed with B6.PPARγloxp/loxp to obtain the second-generation mice. Genomic DNA was extracted from the second-generation mice for PCR to amplify the loxp and Cre gene fragments followed by agarose gel electrophoresis to verify their sizes. The mice with the PPARγloxp/loxp.Nestin-Cre (KO) genotype were selected as the neural stem cell-specific knockout PPARγ mice, with B6.PPARγloxp/loxp (loxp) mice as the control. Tissue samples were collected from specific regions of the mouse brain and peripheral tissue for detecting the expression of PPARγ mRNA using RT-PCR and real-time quantitative PCR.

Genotyping results showed PPARγloxp and Cre bands in the knockout mice, which showed obviously decreased mRNA expression of PPARγ, suggesting successful establishment of neural stem cell-specific PPARγ knockout mice. The two transgenic mice we used were fertile, and their breeding pattern followed the laws of Mendelian inheritance.