Article Figures & Data
Figures
- Figure 1.
FGF-23-deficient mouse model. A, Kaplan–Meier survival curve of WT and FGF-23-deficient (KO) mouse lifespan showing shortened FGF-23-deficient (KO) mouse lifespan (males and females used; n = 10–13/genotype; median survival 63 d, log-rank test ***p < 0.0001). B, During the lifespan study, WT and KO mice were weighed weekly from weaning to death or nine weeks, as relevant. KO mice fail to gain weight after approximately five weeks of age. C, Representative Alizarin red histology of aorta from WT and KO mice. Scale bar represents 100 µm. D, FGF-23 mRNA detected in WT and FGF-23-deficient (heterozygous tissue, HET) adult brain and thymus by ΔΔ Ct qPCR (n = 5; mean ± SEM; ANOVA, ***p < 0.0001). E, Alizarin red staining of WT and KO brain (cerebellum shown; KO lesions indicated by arrows). Graph shows the average number of calcium lesions (five brains/genotype, average number of lesions counted in three bregma-matched sections; t test, **p < 0.005).
- Figure 2.
FGF-23 deficiency impairs spatial discrimination. Separate cohorts of WT, HET, and KO mice were measured at three and five weeks of age. A, B, Open field performance % of time spent along the walls or in the middle of the field and velocity (m/s). C, Context-dependent fear conditioning quantified as % of time spent freezing on training day before shock, training day after foot shock, and testing day, 24 h later when returned to the same context. D, Spatial novelty schematic. Identical objects were explored at a set position (positions a and b) and the following day 1 of the objects was moved to a new position (position c). E, F, Spatial novelty scored as the discrimination index [((time with moved – time with non-moved/total time) × 100); three and five weeks] or % of object interaction time spent with each object, each day (five weeks shown; males and females used; n = 9–11/genotype; mean ± SEM; ANOVA *p < 0.01, **p < 0.005, ****p < 0.0001).
- Figure 3.
FGF-23 deficiency does not affect gross morphology or synaptic plasticity. WT and KO mice measured at six weeks of age. A, Input-output curves plotted as the initial slopes of the evoked field EPSP (fEPSP; mV/ms) as a function stimulus intensity (mV). No change is measured between genotypes. B, Paired-pulse ratio at 50- to 300-ms intervals showing no differences. C, θ Burst induced LTP measured as the fEPSP over time before and after stimulation is not different between genotypes (male mice only for electrophysiology: n = 5–6 mice/genotype, two to four slices per mouse, mean ± SEM; t test). D, Mice injected with 60 mg/kg of PTZ were timed for latency to generalized seizure activity. E, Quantification of total hippocampus (HCX), dentate gyrus, and fimbria volumes (–1.22 to –3.88 mm from bregma). Representative WT and KO Nissl stain. Scale bar is 200 µm (males and females used; n = 8–10 mice/genotype, mean ± SEM; t test).
- Figure 4.
FGF-23 deficiency reduces the number of immature neurons. A, Quantification of all proliferating SGZ cells (Ki67; stereological count) at three and six weeks of age. WT and KO hippocampal neurogenic cell populations as average total cell number across three bregma levels/animal (–1.34 to –2.10 mm) of dorsal hippocampal SGZ. B, Three- and six-week radial-like glial stem cells (BLBP). C, Representative six-week BLBP IHC. D, Percentage of proliferating radial-like glial stem cells (BLBP/Ki67- and BLBP/Ki67+) at six weeks. E, Number of TAPs in six-week-old brain (Sox2+/GFAP-). F, Number of astrocytes (S100β+) in six-week-old SGZ. G, Number of immature neurons (DCX) in three- and six-week-old brain. H, Representative six-week DCX IHC. I, Morphology schematic used to quantify maturation stage of immature neurons at the top of each stage bar graph. Quantification of % of 100 DCX+ immature neurons at each maturation stage. J, Percentage of proliferating immature neurons (DCX/Ki67- and DCX/Ki67+) at six weeks. Scale bars represent 100 or 200 µm; males and females (Lagace et al., 2007) used; n = 6–9/genotype, mean ± SEM; t test: *p < 0.02, **p < 0.002.
Tables
Figure Data structure Statistical test p value 1A Dataset with two groups Log-rank 0.0001
CI: 95%1C Dataset with two groups t test 0.005
CI: 95%1D, left Dataset with two groups t test n.s.
CI: 95%1D, right Dataset with more than two groups One-way ANOVA 0.0001
CI: 95%2A Dataset with more than two groups One-way ANOVA n.s.
CI: 95%2B Dataset with more than two groups One-way ANOVA n.s./0.01
CI: 95%2C Dataset with more than two groups One-way ANOVA n.s./0.005
CI: 95%2D Dataset with more than two groups One-way ANOVA n.s./0.0001
CI: 95%3A Dataset with two groups t test* n.s.
CI: 95%3B Dataset with two groups t test* n.s.
CI: 95%3C Dataset with two groups t test** n.s.
CI: 95%3D Dataset with two groups Log-rank n.s.
CI: 95%3E Dataset with two groups t test* n.s.
CI: 95%4A Dataset with two groups t test* n.s.
CI: 95%4B Dataset with two groups t test* n.s.
CI: 95%4D Dataset with two groups t test* n.s.
CI: 95%4E Dataset with two groups t test 0.02
CI: 95%4F Dataset with two groups t test 0.03
CI: 95%4G Dataset with two groups t test* 0.002
CI: 95%4I Dataset with two groups t test* n.s.
CI: 95%4J Dataset with two groups t test* n.s.
CI: 95%↵Statistical method and p value for all comparisons is detailed by figure, data structure, and statistical test applied. CI indicates confidence interval. When a p value was statistically significant (p < 0.05), the exact p value is reported. Non-significant data are noted as n.s. * denotes that when multiple t tests occurred on a single graph, the relevant condition or age was compared between WT and KO mice tested; ** denotes that the KO was compared to the WT at 0, 60, and 90 min.
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