As the combining target with sperms, ZP3 undergoes an important role in the fertilization of oocytes and therefore it has been the focus in studying the mechanism of mammalian. According to the sequence of the zona pellucida 3 gene of Lagurus lagurus (lZP3), three RNA interference recombinant vectors were constructed with pGenesil-1 aiming at lZP3 mRNA by synthesizing oligonucleotides. And then co-transfected into the Hela cells by Lipofectamine2000 and co-injected into the mice by hydrodynamics-based transfection method with the expression vector pCDNA3-lZP3. In order to select the efficient target sites of lZP3 for RNAi, the mRNA expression level of lZP3 gene in Hela cells and the mouse liver was detected by semi-quantative RT-PCR and real-time PCR. Results show that there are 2 interference vectors can interfere of the expression of lZP3 mRNA, and the mRNAs of the exogenous genes expressed in the mouse liver are coincident with those of in Hela cells after co-transfected with the interference vectors and expression vector. It also suggests that the mice can be the experimental materials for selecting the efficiency target sites of the RNA interference.
Animals ; Arvicolinae ; genetics ; Female ; Gene Expression Regulation ; HeLa Cells ; Humans ; Liver ; metabolism ; Mice ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; Receptors, Cell Surface ; deficiency ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

Leptin receptor deficiency causes morbid obesity and hyperlipidemia in mice. Since physical exercise enhances energy expenditure, it is an important part of successful weight-control regimens. We investigated the mechanism by which swim training regulates leptin receptor deficiency-induced obesity and lipid disorder in a mouse model of obesity (obese db/db mouse). Swim training for 6 weeks significantly decreased body weight gain and adipose tissue mass in both sexes of obese and lean mice, compared to their respective sedentary controls. These effects were particularly evident in obese mice. Swim training also caused significant decreases in serum levels of triglycerides, free fatty acids and total cholesterol in both obese and lean mice. In obese mice, swim training increased the levels of mRNAs and proteins encoding uncoupling protein 1 (UCP1), UCP2 and UCP3 in brown adipose tissue, white adipose tissue and skeletal muscle, respectively. In conclusion, these findings suggest that, in mice, swim training can effectively prevent body weight gain, adiposity and lipid disorders caused by leptin receptor deficiency, in part through activation of UCPs in adipose tissue and skeletal muscle, which may contribute to alleviating metabolic syndromes, such as obesity, hyperlipidemia and type 2 diabetes.
Adipose Tissue/metabolism ; Animals ; Body Weight ; Female ; Ion Channels/genetics/*metabolism ; Lipid Metabolism ; Male ; Mice ; Mitochondrial Proteins/genetics/*metabolism ; Muscle, Skeletal/metabolism ; Obesity/genetics/*metabolism/prevention & control ; *Physical Conditioning, Animal ; RNA, Messenger/metabolism ; Receptors, Cell Surface/*deficiency/genetics/*metabolism ; Receptors, Leptin ; Swimming