Fig. 3

Ovarian morphology and function are unaltered despite the lack of ADGRA3, and estrogen treatment fails to induce vaginal opening in Adgra3^−/− mice. a Representative HE-stained sections of ovaries harvested from 10-week-old mice. Primary and secondary follicles and corpus luteum (CL) formation were observed in all Adgra3^+/+ (n = 3), Adgra3^−/− (n = 3) in proestrus, and Adgra3^−/−CV (n = 3) mice analyzed. b Plasma estradiol levels in 8- to 10-week-old Adgra3^+/+ (n = 5), Adgra3^−/− (n = 3), and Adgra3^−/−CV (n = 3) female mice. Limit of detection (LOD) = 4.04 pmol/L. c Relative mRNA expression of aromatase, Cyp19a1, in ovaries of 8- to 10-week-old Adgra3^+/+ (n = 7), Adgra3^−/− (n = 4), and Adgra3^−/−CV (n = 4) mice. d Relative mRNA expression of estrogen receptor α (Esr1) in the vagina of 8- to10-week-old Adgra3^+/+ (n = 6), Adgra3^−/− (n = 4), and Adgra3^−/−CV (n = 3) mice. e Pre-pubertal external genitalia of Adgra3^+/+ and Adgra3^−/− mice at P15 (vehicle-injected). f Left: Genitalia of Adgra3^+/+ and Adgra3^−/−CV at P15 after estrogen treatment. Right: Representative HE-stained sections of the distal vagina of Adgra3^+/+ and Adgra3^−/−CV at P15 after estrogen treatment (n = 5 per group). Arrow indicates an opened vaginal opening, and an arrowhead indicates a lack of vaginal opening observed in 55% of Adgra3^−/− females after estrogen treatment. V, vagina. g Bone mineral density (BMD) measured in femurs harvested from Adgra3^+/+ (n = 4) and Adgra3^−/− (n = 5) mice at 1 year of age. For ethical reasons, only bones from mice with an open vagina were investigated at year 1. Each dot represents one animal in all graphs. Data are presented as mean ± SEM, and significance was calculated by one-way ANOVA post hoc Dunnett’s multiple comparisons test for three groups and unpaired t-test of two groups. *p = 0.0198