# Gonadal hormones contribute to sex differences in behavior, pathology and epigenetic modifications in the 3×Tg-AD mouse model of Alzheimer’s disease

## TL;DR

Removing sex hormones in male Alzheimer's mice improves memory and reduces disease signs, while in females it worsens memory and increases disease-related gene activity.

## Contribution

This study reveals sex-specific effects of gonadal hormones on Alzheimer's pathology and epigenetic regulation in a mouse model.

## Key findings

- Loss of male gonadal hormones improves spatial learning and reduces amyloid-beta levels in Alzheimer's mice.
- Female gonadal hormone removal impairs learning and increases expression of AD-related genes without affecting amyloid-beta.
- Sex differences in AD pathology are linked to changes in histone variant MacroH2A1 binding at specific genes.

## Abstract

Sex-dependent differences in prevalence and severity are characteristics of Alzheimer’s disease (AD). Using the 3×Tg-AD mouse model, we previously reported that adult males show early behavioral dysfunction, altered epigenetic factors and lack of plaque/tangle pathology. Conversely, adult females retain more severe AD-like pathology and behavior. The present study examines whether gonadal hormones play a role in these differences in current cohorts of 3×Tg-AD mice.

3×Tg-AD and wild-type mice were gonadectomized or sham-operated at 3 months of age. After behavioral phenotyping at 6 months of age, the animals were assessed for molecular markers of AD pathology and expression of genes and histone variants associated with neurodegeneration.

In female transgenic (AD) mice, gonadectomy resulted in poorer spatial learning performance. In contrast, in transgenic male animals, gonadectomy improved spatial learning and memory. Compared to sham-operated AD females, gonadectomized AD females exhibited enhanced expression of mouse (m) Mapt and App genes, consistent with reduced binding activity of the repressive histone variant macroH2A1 at the mMapt gene, but there was no effect on Aβ42 or pTau181 levels. In contrast, gonadectomized AD males showed significantly increased macroH2A1 binding at the mPsen1 promoter, reduced expression of the App and MacroH2A1 genes, and reduced cortical soluble Aβ42 levels compared to sham-operated AD males.

In sum, the results suggest that reduction in serum levels of female gonadal hormones impairs spatial learning capacity, whereas loss of male gonadal hormones enhances spatial learning and memory. In females, gonadectomy reduces binding of the repressive histone variant MacroH2A1 to the mouse Mapt gene and increases expression of the mouse App and Mapt genes without affecting Aβ42 or pTau181 levels. Conversely, loss of male gonadal hormones increases binding of MacroH2A1 to the mouse Psen1 gene and decreases App expression and Aβ42 levels but has no effect on tau expression. Our work suggests that adult gonadal hormones contribute to sex differences in AD-like pathology and performance in learning and memory tasks. Moreover, sex-specific differences in AD-like pathology are partially due to the action of histone variants associated with neurodegeneration, such as macroH2A1.

More women than men have Alzheimer’s disease, and it tends to be more severe in women - are sex hormones responsible?

We compared what happens to male and female mice when sex hormones are removed.

Removing sex hormones led to different results in male and female mice. Loss of sex hormones in males improved learning and memory and lowered amyloid-beta levels (a protein linked to Alzheimer’s disease). But in females, loss of sex hormones led to learning problems and did not affect amyloid-beta. These differences were linked to changes in how certain proteins attach to the DNA, which altered the amount of amyloid-beta differently in male versus female brains.

These results help us understand how Alzheimer’s disease works differently in males and females. This knowledge can be the basis for developing different treatments for males and females.

Loss of gonadal hormones is protective for spatial learning and memory and AD-like pathology in male 3xTg-AD (Alzheimer’s disease) model mice but is detrimental in females.Gonadectomy reduces binding of the repressive histone variant MacroH2A1 to the female mouse Mapt gene, consistent with increased expression of this gene in female, but not male, 3xTg-AD mice.In males only, gonadectomy increases repressive MacroH2A1 binding at the mouse Psen1 gene, reduces expression of the mouse App gene, and reduces cortical soluble Aβ42.Loss of gonadal hormones may trigger sex-specific epigenetic changes, including alterations in histone variants such as MacroH2A1, that contribute via different mechanisms to sex-specific differences in pathology and behavior in Alzheimer’s disease.

Loss of gonadal hormones is protective for spatial learning and memory and AD-like pathology in male 3xTg-AD (Alzheimer’s disease) model mice but is detrimental in females.

Gonadectomy reduces binding of the repressive histone variant MacroH2A1 to the female mouse Mapt gene, consistent with increased expression of this gene in female, but not male, 3xTg-AD mice.

In males only, gonadectomy increases repressive MacroH2A1 binding at the mouse Psen1 gene, reduces expression of the mouse App gene, and reduces cortical soluble Aβ42.

Loss of gonadal hormones may trigger sex-specific epigenetic changes, including alterations in histone variants such as MacroH2A1, that contribute via different mechanisms to sex-specific differences in pathology and behavior in Alzheimer’s disease.