Fig. 6: Inhibition APLN improves cell junction protein expression in human cultured testis. | Nature Communications

Fig. 6: Inhibition APLN improves cell junction protein expression in human cultured testis.

From: Targeting APLN/APJ restores blood-testis barrier and improves spermatogenesis in murine and human diabetic models

Fig. 6

a Immunofluorescence of SOX9 (red) in human testis culture in Day 0 and Day 7 paraffin sections. Scale bar, 20 μm. The percentage of SOX9-positive cells was calculted on Day 0 and Day 7 separately. Mean ± SEM. ns, not significant, unpaired two-tailed t test. n = 3 human testis culture examined over 3 independent experiments. b Immunofluorescence of SYCP3 (green) and CREM (red) in human testis culture on Day 0 and Day 7 paraffin sections. Scale bar, 20 μm. The percentage of SYCP3-positive cells was counted. Mean ± SEM. Unpaired two-tailed t test. n = 3 human testis culture examined over 3 independent experiments. c Bright field diagram of human testis culture between different groups. Scale bar, 50 mm. d, e Immunofluorescence of TJP1 and GJA1 (green) and VIM (red) in human testis culture in Day 7 paraffin sections between indicated sample groups. n = 3 per group. Scale bar, 20 μm. Box-and-whisker plots denote the maximum (top whisker), 75th (top edge of box), 25th (bottom edge of box), and minimum (bottom whisker) percentiles, and the median (line in box). Quantitative analysis of TJP1. Two-tailed student’s t test was performed. f Immunofluorescence of TJP1 or GJA1 (green) and VIM (red) in diabetic patient testis culture in Day 7 paraffin sections between indicated sample groups. Scale bar, 50 μm. n = 3 per group. Box-and-whisker plots denote the maximum (top whisker), 75th (top edge of box), 25th (bottom edge of box) and minimum (bottom whisker) percentiles, and the median (line in box). Quantitative analysis of TJP1 and GJA1. Two-tailed student’s t test was performed. g Hypothetical Mechanism. Elevated blood glucose in diabetic patients directly leads to elevated ROS in Sertoli cells, which promotes HIF1A nuclear translocation and activates Apln expression. The excess of APLN disrupted the BTB-related genes by decreasing NAD+, carnitine, and glutathione. Blocking APLN/APJ with F13A and ML221 could significantly ameliorate the BTB damage and improve low sperm quality.

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