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[Establishment of a mouse model of testicular Occludin gene knockout based on the CRE/LOXP system]
Shun-Chang Pu1,2, Yu Zhang1,2, Ning Lu1,2
1Department of Biology and Food Engineering, Bozhou College, Bozhou, Anhui 236800, China.
Objective:
To establish a testicular Occludin gene knockout model in mice and observe the phenotypic changes.
Methods:
Occludin-floxed (genotype Floxp/-) mice were constructed based on the Cre/loxp system, which were cross-bred with AQP2-cre (genotype Cre/-) mice to derive Occludin knockout mice (Genotype Floxp/Floxp Cre/-). The genotype of the F1 knockout mice was identified by PCR and Southern blot technology. The expression of the Occludin protein in the knockout mice was determined by qPCR, Western blot and immunohistochemistry to verify the success of the modeling. Comparisons were made in the sperm count between the model and normal mice, followed by analysis of their fertility.
Results:
The target mice of Occludin knockout were successfully constructed, which, compared with the normal controls, showed significantly down-regulated expression of the occludin protein, decreased sperm count and reduced fertility (P < 0.05).
Conclusions:
Occludin gene knockout mice were successfully constructed, and deletion of Occludin affects the reproductive function of the mice.
