Sex–specific Avp transcript density in the brain.

(A) Avp transcript density in main brain regions detected by RNAscope. Female 3V region and hypothalamus and male hypothalamus had the highest Avp transcript density. (B) Avp transcript density in nuclei, sub–nuclei and regions of the hypothalamus and forebrain (C) Representative micrographs of some of the hypothalamic and forebrain regions with highest Avp expression. Scale bar: 20 µm. (D) Novel Avp transcripts found in nuclei, sub–nuclei and regions of the hippocampus, thalamus and cerebral cortex. Scale bar: 10 µm. Note, that Avp transcripts were found only in the female 3V ependymal layer and thalamus. N=3 mice per sex. ***P<0.001 and **P<0.01.

Sex–specific Avpr1a transcript density in the brain.

(A) Probe specificity was established by a positive signal in renal tubules of the kidney (positive control) with an absent signal in the frontal association cortex (FrA) (negative control). Sale bar: 20 µm. (B) Representative micrographs of medullary central canal (CC) and hypothalamic arcuate nucleus (Arc). (C) Avpr1a transcript density in main brain regions detected by RNAscope. (D) Avpr1a transcript density in nuclei, sub–nuclei and regions of the ventricular regions, hypothalamus, medulla, midbrain and pons, forebrain, olfactory bulb, hippocampus, thalamus, cortex and cerebellum. N=3 mice per sex. Scale bar: 25 µm (controls). ***P<0.001, **P<0.01 and *P<0.05.

Avp and Avpr1a co-localization in the brain.

We found that various nuclei and sub–nuclei exhibited Avp and Avpr1a co-localization in the brain of both sexes. (A) Female and (B) male Avpr1a to Avp ratios in the cortex, forebrain, hippocampus, 3rd ventricular region, thalamus and hypothalamus.

Avp and Avpr1a transcript densities in the pituitary gland.

RNAscope revealed Avp and Avpr1a expression in the posterior pituitary lobe with Avp and Avpr1a transcript densities that were higher in male compared with female mice. N=3 mice per sex.

Sex–specific Avp–positive cells in the brain.

(A) Total numbers of Avp–positive cells in main brain regions detected by RNAscope. (B) Total numbers of Avp–positive cells in nuclei, sub–nuclei and regions of the hypothalamus, hippocampus, forebrain and cortex. Note, that Avp–positive cells were found only in the female 3V ependymal layer and thalamus. Also shown are representative RNAscope micrographs of hypothalamic regions with highest Avp expression; i.e., supraotpic nucleus (SO), paraventricular hypothalamic nucleus, lateral magnocellular part (PaLM), retrochiasmatic area (RCh), supraoptic nucleus, retrochiasmatic part (SOR) and suprachiasmatic nucleus (SCh). N=3 mice per sex. Scale bar: 20 µm. ***P<0.001 and *P<0.05.

Sex–specific Avpr1a transcript numbers in the brain.

(A) Tolal Avpr1a transcript numbers in main brain regions detected by RNAscope. (B) Avpr1a transcript density in nuclei, sub–nuclei and regions of the medulla, hypothalamus, cortex, midbrain and pons, forebrain, thalamus, cerebellum, hippocampus, olfactory bulb and ventricular regions. Also shown are representative RNAscope micrographs of medullary regions with highest Avpr1a expression; i.e., hypoglossal nucleus (12N), intermediate reticular nucleus (IRt), lateral reticular nucleus (LRt) and medullary reticular nucleus, ventral part (MdV). N=3 mice per sex. Scale bar: 20 µm. ***P<0.001, **P<0.01 and *P<0.05.