In Panel A, the proportion of small (early) growing follicles is plotted against the number of remaining resting follicles (RF) per ovary.

<p>In addition, mean changes in the proportion of resting follicles with at least one close neighbor with respect to age are shown in Panel B, while the proportion of resting follicles (RF) with at least one neighbor are plotted against the number of remaining resting follicles (RF) in Panel C. In addition, in Panel D the correlation between the proportion of early growing follicles and the percentage of follicles with close neighbors is displayed. In all figures, data from WT and Kiss1r KO mice are shown, as blue and red dots, respectively. Yet, regression analyses were applied only to WT data. Similarly, the confidence intervals (CI, presented as blue dotted lines) and the predictive intervals (PI, presented as grey dotted lines) were calculated for the regression slopes obtained from WT individuals. In Panel E, a schematic figure is presented to illustrate of the method used to evaluate the presence of resting follicle neighbors (at a distance of ≤ 60 μm) for resting and early growing follicles. An early growing follicle (GF) lacking close neighbors is highlighted in green, whereas each of the four resting follicles (RF), which have each three neighbors, is marked in red. In Panel F, a distribution plot of frequencies of the number of neighbors in resting and early growing follicles is presented for 3 moth-old WT mice. Data represent the mean ± SEM for n = 5/group. Finally, in Panel G, the Morisita’s index of clustering in WT and Kiss1r KO mice is shown. Values above the dotted line indicate clustered spatial pattern. Different superscript letters indicate statistically significant differences between age-points within each genotype (P<0.05 ANOVA followed by Student-Newman-Keuls test); *, P<0.05 vs. corresponding WT values at the same age-point (Student t-test). Regression analyses, including r² and P values, as well as CI and PI calculations, were conducted using Graph-Pad Prism software. DF: Degrees of freedom.