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. 2017 Jul 28;9(10):1379–1397. doi: 10.15252/emmm.201607376

Figure 4. Lack of Klb in female mice leads to defects in sexual maturation with delay of puberty, altered cyclicity, reduced LH secretion, and reduced fertility.

Figure 4

  • A–C
    Altered pubertal development in KlbKO mice compared to WT littermates including delayed vaginal opening (KO: n = 12; WT: n = 11) and first estrous (KO: n = 7; WT: n = 11), expressed as cumulative percentage as a function of age (Gehan–Breslow–Wilcoxon test), and reduced body weight (KO: n = 7; WT: n = 7; two‐way RM ANOVA).
  • D
    Body weight of adult KlbKO mice (n = 10) compared to WT littermates (n = 15), using unpaired t‐test.
  • E
    Body composition expressed as percentage of fat mass in KlbKO (n = 20) and WT mice (n = 23), unpaired t‐test.
  • F
    Representative low magnification picture of the uterus and ovaries in WT and KlbKO mice. Scale bar 1 mm.
  • G
    Hematoxylin–eosin‐stained ovarian representative sections of KlbKO and WT mice at 3 months (CL: corpora lutea). Scale bar: 600 μm.
  • H
    Normalized ovarian weight expressed as ovary/body weight ratio in KlbKO (n = 7) and WT mice (n = 10), unpaired t‐test.
  • I
    Quantification of corpora lutea per ovary in KlbKO (n = 5) and WT (n = 5) adult females, unpaired t‐test.
  • J
    Representative estrous cycle patterns of 3‐ to 4‐month‐old WT and mutant females, demonstrating marked alteration in KlbKO.
  • K
    Quantification of time spent in different estrous cycle phases in KlbKO (n = 13) and WT (n = 17) adult females, unpaired t‐test.
  • L
    Fertility evaluated as number of litters per female and litter size in KlbKO (n = 6) and WT littermates (n = 5) in a continuous mating protocol (7 months), unpaired t‐test.
  • M
    KlbKO females display blunted preovulatory LH surge in male‐induced proestrous (MIP) test (WT Di: n = 5; WT Pro: n = 8; KO Di: n = 5; KO Pro: n = 5).
  • N
    GnRH test: basal blood LH levels and 30 min after intraperitoneal GnRH injection (0.25 μg) show LH release in both KlbKO (n = 5) and WT littermates (n = 7).
  • O
    Kisspeptin test: blood LH levels at basal state and 30 min after intraperitoneal Kisspeptin injection (Kp‐10; 1 nmole) in WT (n = 5) and KlbKO (n = 5) female mice.
Data information: Multiple comparison analysis of LH levels was performed using a two‐way ANOVA followed by Fisher's LSD test (for MIP) and Sidak's multiple comparison test (for GnRH and Kisspeptin tests). Values shown are mean ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001. ns: not significant.