Fig. 6

DD disturbed Mitochondrial Function in Leydig cells during maturation. Increased mitochondrial membrane potential (Δψm) in DD Leydig cells, assessed by TMRE fluorescence (A). Decreased mitochondrial mass in Leydig cells from DD adult rats, measured using MitoTracker staining (B). The Clustered Image Map datasets display R values from Spearman correlation analyses between blood androgen levels and either Δψm or mitochondrial mass under LD and DD light conditions. Spearman correlation coefficients are shown in the box below: blue indicates a positive correlation with blood androgens, while red indicates a negative correlation (C). DD changed ATP content in Leydig cells isolated from rats of different ages (D). Relative expression of genes involved in mitochondrial biogenesis (E-I): Ppargc1a, Tfam, Nrf1, Cytc, and Cox4/2. Genes regulating mitochondrial fusion (J-M): Opa1, Mfn1, and Mfn2. Genes involved in mitochondrial fission (N, L): Fis1 and Drp1. Markers of mitophagy (O, P): Pink1 and Prkn. Whisker plots represent individual data points (n = 4–5), with lines connecting medians across age groups. Data is expressed as fold change relative to control values at 35 pnd. Statistical significance: #p < 0.05, Kruskal-Wallis test among LD age groups; *p < 0.05, Mann-Whitney test between LD and DD groups at each age point; +p < 0.05, indicating significance in the correlation analysis presented in the cluster image map.