Overexpression of Tfap2a in Mouse Oocytes Impaired Spindle and Chromosome Organization

• Published in: International journal of molecular sciences 2022-11, Vol.23 (22), p.14376
• Main Authors: Lin, Juan, Ji, Zhuqing, Di, Zhengyang, Zhang, Yeqing, Yan, Chen, Zeng, Shenming
• Format: Article
• Language: English
• Subjects: Acetylation; Amino acid sequence; Animals; Antibodies; AP-2 protein; Chromatin; Chromosomes; Chromosomes - genetics; Chromosomes - metabolism; Cytoplasm; Diabetes; Diabetes mellitus (non-insulin dependent); Diabetes Mellitus, Type 2 - metabolism; DNA methylation; DNA-binding protein; Gametocytes; Gene expression; histone acetylation; Histone H3; Histone H4; histone lactylation; Histones; Histones - metabolism; Immunoblotting; Immunofluorescence; Localization; Lysine; Lysine - metabolism; Mammals; Maturation; Meiosis; meiotic maturation; Metaphase; Mice; Microinjection; mouse oocyte; Myc protein; Nucleotide sequence; Oocytes; Oocytes - metabolism; Ovaries; Proteins; Spindles; Tfap2a; Transcription Factor AP-2 - genetics; Transcription Factor AP-2 - metabolism
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms232214376

Transcription factor AP-2-alpha (Tfap2a) is an important sequence-specific DNA-binding protein that can regulate the transcription of multiple genes by collaborating with inducible viral and cellular enhancer elements. In this experiment, the expression, localization, and functions of Tfap2a were investigated in mouse oocytes during maturation. Overexpression via microinjection of Myc-Tfap2a mRNA into the ooplasm, immunofluorescence, and immunoblotting were used to study the role of Tfap2a in mouse oocyte meiosis. According to our results, Tfap2a plays a vital role in mouse oocyte maturation. Levels of Tfap2a in GV oocytes of mice suffering from type 2 diabetes increased considerably. Tfap2a was distributed in both the ooplasm and nucleoplasm, and its level gradually increased as meiosis resumption progressed. The overexpression of Tfap2a loosened the chromatin, accelerated germinal vesicle breakdown (GVBD), and blocked the first polar body extrusion 14 h after maturation in vitro. The width of the metaphase plate at metaphase I stage increased, and the spindle and chromosome organization at metaphase II stage were disrupted in the oocytes by overexpressed Tfap2a. Furthermore, Tfap2a overexpression dramatically boosted the expression of p300 in mouse GV oocytes. Additionally, the levels of pan histone lysine acetylation (Pan Kac), histone H4 lysine 12 acetylation (H4K12ac), and H4 lysine 16 acetylation (H4K16ac), as well as pan histone lysine lactylation (Pan Kla), histone H3 lysine18 lactylation (H3K18la), and H4 lysine12 lactylation (H4K12la), were all increased in GV oocytes after Tfap2a overexpression. Collectively, Tfap2a overexpression upregulated p300, increased the levels of histone acetylation and lactylation, impeded spindle assembly and chromosome alignment, and ultimately hindered mouse oocyte meiosis.