Figure 3. Functional neuronal types in VNOs from male and female mice.
(A &B) Cell counts in each male (×) and female (○) VNO imaging volume, in terms of total number of steroid-responsive cells (a) and number of cells of each VSN functional type (B). Type 8 were more abundant in male than in female mice (p=3.1×10−5, Students’ t-test, significance tested with Ŝidák correction for multiple comparisons). ***: p<0.001; **: p<0.01; *: p<0.05. Note type 6 and 11 were not significant when corrected for multiple comparisons. Throughout the figures involves multiple cell types, y axes are in logarithmic scale, and “# of cells +1” were plotted to avoid displaying log0=−∞.
(C &D) Three-dimensional images (upper) and two-dimensional slices (lower) of male (C) and female (D) VNOs. Red indicates the fluorescence change caused by epitestosterone sulfate (A6940) (ΔF/FA6940), and green colorizes max(ΔF/FA7010, ΔF/FP8200). Type-8 cells, exclusively responsive to A6940, are therefore red; note these were present only in the male VNO but not in the female VNO. Responses induced by P8200 (green) were particularly intense and spread to neighboring pixels, producing a green halo around the yellow cells. Scale bar, 50 μm.
(E) Type-8 VSNs were found in all 6 male VNO imaging volumes (from 4 individual male mice). Dashed lines separate cells from different imaging volumes. Each imaging volume is labeled with the animal’s identity; multiple imaging volumes collected from the same animal are differentiated by letter.
(F) Variability of male (×) and female (○) mice. For each sex, cell count variability of each type, and 95% confidence interval for multinomial random sampling, are shown as in Figure 2B. The confidence intervals are shown in pink and blue for the female and male groups, respectively. Within-sex variabilities were not distinguishable from random sampling in 3 (male) and 11 (female) cell types.
See also Figure S3 & S4, and Movie S2–4.