Breeding and genotype identification of NLRP3 gene knock-out mice / 第二军医大学学报

Objective: To breed and identify NLRP3 gene knock-out mice. Methods: The NLRP3 gene knock-out heterozygote mice were bred alone and copulated. The offsprings were to have three genotypes: wild genotype, heterozygote genotype and homozygote genotype. Genomic DNA was obtained from each pups and were subjected to PCR and T7 endonuclease 1 to identify the genotype. The homozygote mice were mated with the opposite sex heterozygote mice to obtain more homozygote pups. Results: Breeding and reproducing were both successful, and we obtained heterozygote genotype and homozygote genotype mice with NLRP3 gene knock-out. Conclusion: Correct methods of breeding, reproducing and identifying can effectively obtain NLRP3 gene knock-out mice from heterozygote mice.

Construction of Rtn4-A/B knockout mousemodel / 第二军医大学学报

Objective To generate Rtn4-A/B knockout mouse model and to explore the biological function of the Rtn4-B gene. Methods The targeting construct for inactivating Rtn4-A/B gene was prepared by bacterial artificial chromosome (BAC). The vector was linearized and electroporated into 129SvEv mouse embryonic stem cells (ES cells). Then the Rtn4-A./B knockout ES cells weremicroinjected into blastula of C57BL/6J mice after superovulation. F1 hybrid mice were bred to obtain mouse aggregation chimeras, and were identifiedby PCR amplification of tail genomic DNA. Results Fourteen clones of gene-targeted ES cells were identified after gene knockout and five male chimeras with a higher than 50 chimeric ratio were produced after microinjection into the blastula. Finally four Rn4-A/B hybrid mice were obtained. Conclusion A Rtn4-A/B deficient mouse strainhas been successfully generated by homologous recombination using genetically modified ES cells.

Changes of peripheral pain threshold in transient receptor potential vanilloid 1 knockout female mice / 第二军医大学学报

Objective To investigate the changes of peripheral pain threshold after knockout of transient receptor potential vanilloid 1(TRPV1) gene. Methods Tail-Flick Analgesia Meterand von-Frey hair were used to determine the peripheral thermal and mechanical thresholds in TRPV1 knockout and wild-type femalemice, and the results of the two groups were compared. Results The Tail-Flick latency in TRPV1 knockout mice was significantly prolonged after hot stimulation compared with that in thewild-type group([3. 59±0. 65] s vs [2. 19±0. 24] s, P0. 05). Conclusion It is suggested that TRPV1 receptor mediate thermal stimuii response under physiological condition, and has no noticeable influence on mechanical stimuii response.

Changes of peripheral pain threshold in transient receptor potential vanilloid 1 knockout female mice / 第二军医大学学报

To investigate the changes of peripheral pain threshold after knockout of transient receptor potential vanilloid l (TRPVl) gene. Methods Tail-Flick Analgesia Meterand von-Frey hair were used to determine the peripheral thermal and mechanical thresholds in TRPVl knockout and wild-type female mice, and the results of the two groups were compared. Results The Tail-Flick latency in TRPVl knockout mice was significantly prolonged after hot stimulation compared with that in the wild-type group([3.59±0.65] s vs [2.l9±0.24] s, P0.05). Conclusion It is suggested that TRPVl receptor mediate thermal stimuli response under physiological condition, and has no noticeable influence on mechanical stimuli response.