Fig. 4: SEMA6A loss delays puberty and gonadal maturation in female and male mice.
From: SEMA6A drives GnRH neuron-dependent puberty onset by tuning median eminence vascular permeability
a Age at VO in adult females of indicated genotypes (Sema6a+/+ n = 15; Sema6a+/- n = 17; Sema6a-/- n = 9. Sema6a+/- p = 0.9376, Sema6a-/- p < 0.0001 and delayed Sema6a-/- only p < 0.0001 vs. Sema6a+/+). Red dots indicate Sema6a-/- females without VO at sacrifice. b Ages at VO and FE in adult females of indicated genotypes (Sema6a+/+ n = 9; Sema6a+/- n = 18; Sema6a-/- n = 9. VO: Sema6a+/- p = 0.5186 and Sema6a-/- p = 0.0002 vs. Sema6a+/+. FE: Sema6a+/- p = 0.5550 and Sema6a-/- p < 0.0001 vs. Sema6a+/+). Red triangle indicates Sema6a-/- female without FE at sacrifice. c Age at BPS in adult males of indicated genotypes (Sema6a+/+ n = 11; Sema6a+/- n = 22; Sema6a-/- n = 14. Sema6a+/- p = 0.0615 and Sema6a-/- p < 0.0001 vs. Sema6a+/+). Red dots indicate Sema6a-/- males without BPS at sacrifice. d Quantification of ME innervation (as in e, f) in adult female and male brains at VO/BPS revealed a significant decreased GnRH staining in Sema6a-/- compared to Sema6a+/+ in both sexes (females: n = 3 per group; p = 0.0311 males: n = 5 per group; p = 0.0389). Red dots indicate Sema6a-/- females without VO. Coronal sections of female (e) and male (f) ME immunolabelled for GnRH. Presence of GnRH neuron axon terminals in Sema6a+/+ brains (black arrows) is reduced in Sema6a-/- (Δ). g–i Weight (g) and H&E histological analysis (h) of adult female ovaries. Sema6a-/- mice exhibited significantly smaller ovaries (Sema6a+/+ n = 20, Sema6a-/- n = 12; p = 0.0049) but normal folliculogenesis (i, n = 6 per group; primary p = 0.7666, secondary p = 0.6837, pre-antral p = 0.3500, early antral p = 0.8205). Red dot indicates Sema6a-/- female without FE. j–l Weight (j) and immunostaining for Leydig cell marker CYP17A1 (k) of adult male testes. Sema6a-/- mice exhibited significant smaller testes (Sema6a+/+ n = 10, Sema6a-/- n = 12) and reduced number of Leydig cell evaluated as CYP17A1+ area (l, n = 6 per group; p = 0.0002). Red dots indicate Sema6a-/- males without BPS at sacrifice. m RT-qPCR analysis for Lhb transcript in male pituitaries (Sema6a+/+ n = 7, Sema6a-/- n = 9; p = 0.0161) calculated relative to controls using Gapdh-normalized Cq threshold values. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 and NS (not significant) after One- (a, c) or Two-way (b) ANOVA followed by Dunnett’s post-hoc test, or Two-tailed unpaired Student’s t test (d, g, i, j, l, m) Abbreviations: VO vagina opening, FE first estrous, BPS balanopreputial separation, 3v third ventricle, del. delayed. Scale bars: 500 μm (h), 250 μm (e, f, k). Data are presented as mean ± SD. Source data are provided as a Source Data file.
