Abstract
OBJECTIVE: To construct a HIV-1 gp120 transgenic mouse model (gp120(+)) with α7 nicotinic acetylcholine receptor (α7nAChR) gene knockout. METHODS: The α7nAChR gene knockout mice (α7R(-/-)) were crossed with HIV-1gp120 transgenic mice (gp120(+)) to generate F1 generation mice. We selected the F1 mice with the genotype of α7R(+/-)/gp120(+) to mate to obtain the F2 mice. The genotypes of the F3 mice were identified by PCR, and the protein expressions in the double transgenic animal model was analyzed by immunohistochemistry. BV2 cells were treated with gp120 protein and α7nAChR inhibitor, and the expressions of IL-1β and TNF-α were detected using ELISA. RESULTS: The results of PCR showed the bands of the expected size in F3 mice. Two F3 mice with successful double gene editing (α7R(-/-)/gp120(+)) were obtained, and immunohistochemistry showed that the brain tissue of the mice did not express α7 nAChR but with high gp120 protein expression. In the in vitro cell experiment, treatment with gp120 promoted the secretion of IL-1β and TNF-α in BV2 cells, while inhibition of α7nAChR significantly decreased the expression of IL-1β and TNF-α (P < 0.001). CONCLUSIONS: By mating gp120 Tg mice with α7R(-/-) mice, we obtained gp120 transgenic mice with α7nAChR gene deletion, which serve as a new animal model for exploring the role of α7nAChR in gp120-induced neurotoxicity.