Key events in the process of sex determination and differentiation in early chicken embryos

Objective Current understanding of sex determination and differentiation mechanisms during early chicken embryonic development remains incomplete. To address this, we applied RNA sequencing to identify male-female expression differences at critical developmental stages (E0 blastocysts, E3.5-E6.5 gen...

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Main Authors: Xiaoqian Lv, Changhua Sun, Xin Liu, Guanzheng Liu, Wei Gong, Hongwu Qian, Zeyu Li, Jun Wu, Xilin Zhu, Jiuzhou Song, Yingjie Niu, Hongyan Sun, Wei Han, Guo hong Chen, Kai Jin, Bichun Li, Qisheng Zuo
Format: Article
Language:English
Published: Asian-Australasian Association of Animal Production Societies 2025-06-01
Series:Animal Bioscience
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Online Access:http://www.animbiosci.org/upload/pdf/ab-24-0679.pdf
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author Xiaoqian Lv
Changhua Sun
Xin Liu
Guanzheng Liu
Wei Gong
Hongwu Qian
Zeyu Li
Jun Wu
Xilin Zhu
Jiuzhou Song
Yingjie Niu
Hongyan Sun
Wei Han
Guo hong Chen
Kai Jin
Bichun Li
Qisheng Zuo
author_facet Xiaoqian Lv
Changhua Sun
Xin Liu
Guanzheng Liu
Wei Gong
Hongwu Qian
Zeyu Li
Jun Wu
Xilin Zhu
Jiuzhou Song
Yingjie Niu
Hongyan Sun
Wei Han
Guo hong Chen
Kai Jin
Bichun Li
Qisheng Zuo
author_sort Xiaoqian Lv
collection DOAJ
description Objective Current understanding of sex determination and differentiation mechanisms during early chicken embryonic development remains incomplete. To address this, we applied RNA sequencing to identify male-female expression differences at critical developmental stages (E0 blastocysts, E3.5-E6.5 genital ridges, E18.5 gonads), focusing on glycolysis, histone acetylation, and DNA methylation. This approach aims to unravel key regulatory mechanisms and advance developmental biology insights. Methods We analyzed molecular mechanisms of chicken sex determination at key stages (E0 blastocysts, E3.5-E6.5 genital ridges, E18.5 gonads) using RNA sequencing. Glycolysis, histone acetylation, and DNA methylation levels were assessed in embryonic stem cells and chicken embryonic fibroblasts. E18.5 gonads were treated with glycolytic activators (SB431542 and PD0325901 [2i]), a DNA demethylation activator (Vitamin C [Vc]), or an inhibitors of histone acetylation modification (valproic acid [VPA]). Sex-related gene expression, hormone levels, and gonad morphology were evaluated to determine treatment effects. Results Key findings revealed that sex differences emerged as early as the blastocyst stage, intensified with embryonic development and were marked by a surge in sexually dimorphic gene expression. Gene Ontology and Kyoto encyclopedia of genes and genomes analyses highlighted the pivotal roles of energy metabolism and epigenetic modification process during this critical period. 2i, VC, or VPA interventions targeting E18.5 embryo gonads, induced a remarkable transformation of ovarian tissue into a testis-like structure, characterized by cortical thinning, medulla densification, downregulation of female-specific genes (FOXL2, WNT4), upregulation of male-specific genes (SOX9, AMH), and increased testosterone secretion. This phenotypic and molecular shift underscores the ability of metabolic and epigenetic modulators to reprogram ovarian development towards a male-like pattern, preserving male sexual characteristics. Conclusion Our study establishes energy metabolism and epigenetic regulation as central drivers of avian sex determination. These findings advance understanding of vertebrate developmental biology and provide a framework for dissecting regulatory networks in avian sexual development.
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spelling doaj-art-1a515186935f422d99f8ce624bcc93fa2025-08-20T02:16:10ZengAsian-Australasian Association of Animal Production SocietiesAnimal Bioscience2765-01892765-02352025-06-013861081110410.5713/ab.24.067925423Key events in the process of sex determination and differentiation in early chicken embryosXiaoqian Lv0Changhua Sun1Xin Liu2Guanzheng Liu3Wei Gong4Hongwu Qian5Zeyu Li6Jun Wu7Xilin Zhu8Jiuzhou Song9Yingjie Niu10Hongyan Sun11Wei Han12Guo hong Chen13Kai Jin14Bichun Li15Qisheng Zuo16 Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou, China Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou, China Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou, China Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou, China Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou, China Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou, China Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou, China Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou, China Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou, China Department of Animal & Avian Sciences, University of Maryland, College Park, MD, USA Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou, China Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou, China Jiangsu Institute of Poultry Sciences/Poultry institute, Chinese Academy of Agricultural Sciences, Yangzhou, China Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou, China Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou, China Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou, China Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, Yangzhou, ChinaObjective Current understanding of sex determination and differentiation mechanisms during early chicken embryonic development remains incomplete. To address this, we applied RNA sequencing to identify male-female expression differences at critical developmental stages (E0 blastocysts, E3.5-E6.5 genital ridges, E18.5 gonads), focusing on glycolysis, histone acetylation, and DNA methylation. This approach aims to unravel key regulatory mechanisms and advance developmental biology insights. Methods We analyzed molecular mechanisms of chicken sex determination at key stages (E0 blastocysts, E3.5-E6.5 genital ridges, E18.5 gonads) using RNA sequencing. Glycolysis, histone acetylation, and DNA methylation levels were assessed in embryonic stem cells and chicken embryonic fibroblasts. E18.5 gonads were treated with glycolytic activators (SB431542 and PD0325901 [2i]), a DNA demethylation activator (Vitamin C [Vc]), or an inhibitors of histone acetylation modification (valproic acid [VPA]). Sex-related gene expression, hormone levels, and gonad morphology were evaluated to determine treatment effects. Results Key findings revealed that sex differences emerged as early as the blastocyst stage, intensified with embryonic development and were marked by a surge in sexually dimorphic gene expression. Gene Ontology and Kyoto encyclopedia of genes and genomes analyses highlighted the pivotal roles of energy metabolism and epigenetic modification process during this critical period. 2i, VC, or VPA interventions targeting E18.5 embryo gonads, induced a remarkable transformation of ovarian tissue into a testis-like structure, characterized by cortical thinning, medulla densification, downregulation of female-specific genes (FOXL2, WNT4), upregulation of male-specific genes (SOX9, AMH), and increased testosterone secretion. This phenotypic and molecular shift underscores the ability of metabolic and epigenetic modulators to reprogram ovarian development towards a male-like pattern, preserving male sexual characteristics. Conclusion Our study establishes energy metabolism and epigenetic regulation as central drivers of avian sex determination. These findings advance understanding of vertebrate developmental biology and provide a framework for dissecting regulatory networks in avian sexual development.http://www.animbiosci.org/upload/pdf/ab-24-0679.pdfchickendna methylationglycolysishistone acetylationsex determination
spellingShingle Xiaoqian Lv
Changhua Sun
Xin Liu
Guanzheng Liu
Wei Gong
Hongwu Qian
Zeyu Li
Jun Wu
Xilin Zhu
Jiuzhou Song
Yingjie Niu
Hongyan Sun
Wei Han
Guo hong Chen
Kai Jin
Bichun Li
Qisheng Zuo
Key events in the process of sex determination and differentiation in early chicken embryos
Animal Bioscience
chicken
dna methylation
glycolysis
histone acetylation
sex determination
title Key events in the process of sex determination and differentiation in early chicken embryos
title_full Key events in the process of sex determination and differentiation in early chicken embryos
title_fullStr Key events in the process of sex determination and differentiation in early chicken embryos
title_full_unstemmed Key events in the process of sex determination and differentiation in early chicken embryos
title_short Key events in the process of sex determination and differentiation in early chicken embryos
title_sort key events in the process of sex determination and differentiation in early chicken embryos
topic chicken
dna methylation
glycolysis
histone acetylation
sex determination
url http://www.animbiosci.org/upload/pdf/ab-24-0679.pdf
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