Establishment of eNOS Knockout Rat Model and Phenotypic Study

doi:10.3969/j.issn.1674-5817.2015.04.002

Abstract

Abstract: Objective To establish an endothelial nitric oxide synthase (eNOS) knockout rat model. Methods The encoding eNOS specific Zinc-finger nucleases (ZFN) mRNA was transcripted in vitro and purified, then injected into zygotes. The injected zygotes were transferred into oviducts of pseudopregnant rats. The genotype of knockout rats were identified by PCR and sequencing. The histopathological changes were observed by HE staining. The eNOS expression were detected by Western blot. The body weight differences were detected at 8-16 weeks ages, and the blood pressures, systokic pressures, diastolic pressures and heart rates were determined at 8 weeks age by using a tail-cuff system. Result Total of 441 zygotes were injected and 365 zygote cells were transplanted into oviducts of 12 recipients. Twenty-three offspring were born, and the results of PCR and sequencing showed that 4 offsprings were eNOS knockout rats. The screening result showed No.8 offspring was homozygous, and it exhibited limb defect, abnormal structures of artery. The results of Western blot showed that eNOS were not expressed in the kidney and heart in eNOS knockout rats. The comparison of body weights showed that the effect of eNOS genotype were significant different. Blood pressures, systokic pressures and diastolic pressures were higher in eNOS knockout rats than the SD rats, and there were significant differents. Heart rates were lower in the females eNOS knockout rats, but in the males eNOS knockout rats there were not significantly different. Conclusion eNOS knockout rats have been established successfully. This rat model will contribute to research of hypertension.

Key words: Endothelial nitric oxide synthase (eNOS), Zinc-finger nucleases (ZFN), Rats, Gene knockout, Hypertension

CLC Number:

Q95-33

YANG Wei, ZHOU Sheng-lai, DONG Wan-wei, WANG Wei, YU Yang, GUO Xiao-chong, ZHANG E-nuo, WANG Hong-yu, ZHENG Zhi-hong. Establishment of eNOS Knockout Rat Model and Phenotypic Study[J]. Laboratory Animal and Comparative Medicine, 2015, 35(4): 271-276.

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