# Estrogen receptor regulation of brain sexual differentiation

> **NIH NIH F31** · COLD SPRING HARBOR LABORATORY · 2020 · $29,520

## Abstract

PROJECT SUMMARY/ABSTRACT
In sexually reproducing species, males and females display different social and reproductive behaviors, such as
mating and aggression. These behaviors typically require no training, indicating they are developmentally
programmed in the brain. In many mammalian species, including primates and rodents, sexual differentiation of
the brain is regulated by nuclear receptor transcription factors (TFs), which bind gonadal steroid hormones, such
as testosterone and estrogens. In mice, a perinatal surge of testosterone permanently masculinizes a key brain
region controlling sex-typical behaviors, called the bed nucleus of the stria terminalis (BNST). Within the perinatal
BNST, testosterone is converted to estradiol, which activates estrogen receptor α (ERα). Previous genetic
knockout and pharmacological studies demonstrate perinatal ERα signaling is necessary and sufficient for
masculinization of BNST circuitry and behavior. Specifically, ERα activation leads to a male-bias in BNST cell
survival and axon guidance, particularly to the anteroventral periventricular nucleus (AVPV), between postnatal
day 4 (P4) and P10. This project seeks to identify and characterize ERα genomic binding sites and target genes
involved in BNST sexual differentiation. Recently our group discovered that estradiol regulates the expression
of the netrin receptor, Unc5b, in adult BNST ERα+ cells. Unc5b is robustly expressed in the neonatal BNST and
has previously been shown to regulate neuron survival and axon guidance. Aim 1 of this project investigates
whether estradiol regulation of Unc5b contributes to BNST sexual differentiation, using mice lacking Unc5b
expression in ERα+ cells (Esr1cre/+;Unc5blx/lx). The experiments in Aim 1 test the hypothesis that estradiol
regulation of Unc5b contributes to male-biased BNST cell survival and/or AVPV innervation. Because ERα likely
masculinizes the BNST through multiple biological pathways, Aim 2 of this project seeks to identify the complete
repertoire of ERα genomic binding sites and target genes in the developing BNST. Previously, I discovered the
first ERα genomic binding sites in the adult brain, using a recently published low-input TF profiling method, called
CUT&RUN (Cleavage Under Targets & Release Under Nuclease). This approach revealed brain-specific ERα
binding sites are enriched near genes involved in neurodevelopmental processes. Aim 2 of this application uses
CUT&RUN to measure estradiol-regulated ERα binding sites in the perinatal BNST/hypothalamus. Aim 2 also
measures perinatal estradiol-regulated gene expression in P4 BNST ERα+ cells. Using a CRISPR-mediated
activation (CRISPRa) system in primary neurons, distal ERα binding sites will be causally linked to the
expression of perinatal estradiol-regulated genes. Overall, the project will reveal how nuclear receptors regulate
genes involved in brain sexual differentiation and, in doing so, will provide novel insight into the molecular basis
of sex-bia...

## Key facts

- **NIH application ID:** 10067180
- **Project number:** 1F31MH124365-01
- **Recipient organization:** COLD SPRING HARBOR LABORATORY
- **Principal Investigator:** Jordan Bruno Gegenhuber
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $29,520
- **Award type:** 1
- **Project period:** 2020-09-01 → 2023-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10067180

## Citation

> US National Institutes of Health, RePORTER application 10067180, Estrogen receptor regulation of brain sexual differentiation (1F31MH124365-01). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10067180. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*