# Integrated Analysis of Testicular Histology, Sperm Quality, and Gene Expression (TGFB2, DMRT1) in Rooster Semen (Gallus gallus domesticus)

## TL;DR

This study explores how testicular structure, sperm quality, and gene expression (TGFB2, DMRT1) are related in roosters, suggesting an integrated approach for assessing reproductive health.

## Contribution

The study reveals novel correlations between DMRT1 gene expression and reproductive parameters in roosters, supporting integrated analysis for reproductive assessment.

## Key findings

- DMRT1 gene expression negatively correlates with testicular weight, sperm motility, and viability.
- TGFB2 gene expression shows no significant associations but has a moderate positive correlation with DMRT1 expression.
- Integrated analysis of histomorphology, sperm quality, and gene expression is recommended for reproductive evaluation.

## Abstract

The aim of this study was to identify correlations between testicular histomorphology, fresh semen parameters, and the expression level of key spermatogenesis genes—TGFB2 and DMRT1—in roosters. Analysis of TGFB2 and DMRT1 gene expression in fresh semen demonstrated a close relationship between molecular genetic mechanisms and histomorphometric parameters. Thus, the expression level of the DMRT1 gene, which is key in determining sex in birds during embryogenesis, showed a number of negative correlations with parameters such as testicular weight, total/progressive sperm motility, and viability. TGFB2 gene expression in fresh semen showed no significant association with the studied parameters, but correlation analysis revealed a moderate positive association with DMRT1 gene expression. The results of this study support the need for an integrated approach to assessing the reproductive performance of males and the quality of sperm produced.

The study of the relationship between testicular morphology and sperm quality is a pressing issue, for which molecular genetic approaches, including quantitative analysis of gene expression, are being implemented. The aim of this study was to identify correlations between the histomorphological structure of the testes, fresh sperm parameters, and the expression level of key spermatogenesis genes—TGFB2 and DMRT1—in roosters. The experiment was conducted on 10 Russian Snow White roosters aged 28–32 weeks. Sperm quality was assessed by volume, sperm concentration, total and progressive motility, and viability; histological analysis of the rooster testes was performed. The relative expression of the TGFB2 and DMRT1 genes in sperm was analyzed. Multiple correlation analysis of the data was conducted. A positive correlation was found between ejaculate volume and the number of spermatogonia (p = +0.651), a negative correlation between ejaculate volume and the number of second-order spermatocytes (p = −0.704), a negative correlation between the total cross-sectional area of the seminiferous tubules of the testes and sperm viability (p = −0.782), a negative correlation between the number of seminiferous tubules and the average diameter of their cross-section (p = −0.685), and a positive correlation between total and progressive sperm motility (p = +0.794). Analysis of TGFB2 and DMRT1 gene expression in sperm demonstrated a certain relationship between molecular genetic mechanisms and histomorphometric parameters. The expression level of the DMRT1 gene, which plays a key role in sex determination in birds during embryogenesis, had a number of negative correlations with such parameters as testicle weight (r = −0.782), total/progressive sperm motility (r = −0.552; r = −0.612), and viability (r = −0.552). Expression of the TGFB2 gene had no significant relationship with the studied parameters, but correlation analysis revealed a moderate positive relationship (r = +0.321) with DMRT1 gene expression. The data obtained indicate the expediency of integrating morphometric, cellular, and molecular analysis for an objective assessment of rooster reproductive function.