PMC

Male Vax1 HET mice have reduced sperm count and motility. A, H&E-stained testes. Scale bar, 10 μm. B, Total sperm count per epididymis, C, Percentage of motile sperm (n = 6). D, H&E-stained sperm. Scale bar, 1 μm. Data represent means ± SEM (Student's t test as compared with control). **, P < .01; ***, P < .001.

A, Gene transcript levels in gonadotrope pull-down of pituitary from RiboTag male mice crossed with αGSU-icre⁺ (cre⁺) and αGSU-icre⁻ (cre⁻) mice (n = 4). Gene transcript levels of male Vax1 WT and HET were determined by qRT-PCR of hypothalamus (n = 5–8) (B) and pituitary (n = 5–8) (C). All data were analyzed by Student's t test as compared with control. **, P < .01; ***, P < .001. A.U., arbitrary units.

A, Gene transcript levels in gonadotrope pull-down of pituitary from RiboTag:αGSU-icre⁺ diestrous and proestrous females (n = 2–3, each n represents two animals). Female Vax1 WT and HET gene transcript levels were determined by qRT-PCR of pituitary (n = 6–9) (B), hypothalami (n = 6–9) (C), and ARC and AVPV (n = 6–7) (D). Statistical analysis was by Student's t test as compared with control. *, P < .05; **, P < .01.

Hormone levels in Vax1 WT and HET mice. Serum was collected from adult (A–C) male and (D–G) female diestrus mice and assayed for T, FSH, LH, and E₂. Data represent means ± SEM. Gray line indicates median. Statistical analysis was by Student's t test as compared with WT. *, P < .05; **, P < .01. Male T levels (A) were analyzed by the nonparametric Mann-Whitney test (P = .976).

A, Estrous cycling was monitored daily in Vax1 WT and HET females. B, Time spent in each stage of the cycle. C, Average cycle length was determined (n = 15). D, H&E-stained ovary for histology evaluation. E, Number of CL in the ovary of diestrous females (n = 5–7). Number of total (F) and healthy (G) oocytes in superovulated Vax1 WT and HET females (n = 3–4) is shown. B, Statistical analysis by two-way ANOVA followed by Bonferroni. *, P < .05; **, P < .01; ***, P < .001 as compared with WT in the same stage of the cycle. C and E–G, Student's t test as compared with control. *, P < .05; ***, P < .001.

Vax1 adult mice lose more than 50% of GnRH neurons. Immunohistochemistry of GnRH was performed in WT and Vax1 HET mice. A, Illustrative images of GnRH staining in the male hypothalamus. Arrowheads indicate GnRH neurons. Scale bar, 100 μm. B, Total GnRH neuron counts in the adult mouse brain (n = 3–4). Statistical analysis was by Student's t test as compared with WT of the same gender. *, P < .05. C, Serum LH was measured prior to and 10 minutes after a single GnRH challenge in Vax1 diestrous/metestrous females and males. Statistical analysis by two-way ANOVA, as compared before GnRH. **, P < .01; ***, P < .001.

A, RT-PCR for Vax1 and the housekeeping gene, H2AFz, in various tissues from male and female C57BL/6J mice. B, Fertility assessment of Vax1 HET mice. Virgin inexperienced 10- to 16-week-old male WT and Vax1 HET (WT, HET) mice were cohoused with virgin inexperienced 10- to 16-week-old WT or Vax1 HET female mice and the number of days to first vaginal plug monitored. In a second set of experiments, virgin male and female mice aged 9–11 weeks of age at the start of the experiment were used to determine fertility in Vax1 HETs. C, The number of days to first litter (n = 5–12). D, The average litter size of the first three litters (n = 10–13). E, The number of litters in 3 months were recorded (n = 10–13). Data represent means ± SEM. Statistical analysis was done by Student's t test as compared with WT × WT. *, P < .05; **, P < .01.