Figure 6.

AAV-mediated gene therapy may ensure long-term benefits with a single treatment

(A) Experimental overview of the in vivo studies.

(B) Representative images of LHCGR expression in the testicular interstitium 6 months after treatment in Lhcgr^−/− mice injected with PBS or AAV8-Lhcgr (n = 4). Nuclei were counterstained with DAPI. NC, negative control. Scale bar: 50 μm.

(C and D) The concentrations of serum (C) and intratesticular (D) testosterone were analyzed 6 months after treatment in 3-week-old Lhcgr^−/− mice injected with PBS or AAV8-Lhcgr (n = 5).

(E) Representative images of CYP17A1expression in the testicular interstitium in Lhcgr^−/− mice injected with PBS or AAV8-Lhcgr (n = 4). Nuclei were counterstained with DAPI. NC, negative control. Scale bar: 50 μm.

(F and G) Representative photographs of the external (F) and internal genitalia (G) of Lhcgr^−/− mice injected with PBS or AAV8-Lhcgr (n = 4). Arrows (F) and arrowheads (G) indicate the testes. Scale bar: 1 cm.

(H and I) Representative light micrographs of testes (H) and epididymis (I) sections from Lhcgr^−/− mice injected with PBS or AAV8-Lhcgr (n = 4). Arrows indicate Leydig cells, and arrowheads indicate full spermatogenesis in the testis (H). Stars indicate spermatozoa in the cauda epididymis (I). Scale bars: 100 μm.

(J) Representative images of testicular sections from Lhcgr^−/− mice injected with PBS or AAV8-Lhcgr (n = 4). Sections were immunostained for DDX4 and TNP2 and counterstained with DAPI. Scale bars: 50 μm.

(K–M) The sperm counts (K) and proportions of sperm with motility (L) and progressive motility (M) were analyzed in Lhcgr^−/− mice injected with PBS or AAV8-Lhcgr (n = 4). Data are represented by boxplots, and whiskers show the minimum to maximum values. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.