Molecular Mechanism of m6A Methylation Modification Genes <i>METTL3</i> and <i>FTO</i> in Regulating Heat Stress in Sheep
Heat stress is an important environmental factor affecting livestock production worldwide. Primary hepatocytes and preadipocytes derived from Hu sheep were used to establish a heat stress model. Quantitative reverse transcriptase-PCR (qRT-PCR) analysis showed that heat induction significantly increa...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-07-01
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| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/24/15/11926 |
| Summary: | Heat stress is an important environmental factor affecting livestock production worldwide. Primary hepatocytes and preadipocytes derived from Hu sheep were used to establish a heat stress model. Quantitative reverse transcriptase-PCR (qRT-PCR) analysis showed that heat induction significantly increased the expression levels of heat stress protein (HSP) genes and the N<sup>6</sup>-methyladenosine (m6A) methylation modification genes: methyltransferase-like protein 3 (<i>METTL3</i>), methyltransferase-like protein 14 (<i>METTL14</i>), and fat mass and obesity associated protein (<i>FTO</i>). Heat stress simultaneously promoted cell apoptosis. Transcriptome sequencing identified 3980 upregulated genes and 2420 downregulated genes related to heat stress. A pathway enrichment analysis of these genes revealed significant enrichment in fatty acid biosynthesis, degradation, and the PI3K-Akt and peroxisome proliferator-activated receptor (PPAR) signaling pathways. Overexpression of <i>METTL3</i> in primary hepatocytes led to significant downregulation of <i>HSP60</i>, <i>HSP70</i>, and <i>HSP110</i>, and significantly increased mRNA m6A methylation; <i>FTO</i> interference generated the opposite results. Primary adipocytes showed similar results. Transcriptome analysis of cells under <i>METTL3</i> (or <i>FTO</i>) inference and overexpression revealed differentially expressed genes enriched in the mitogen-activated protein kinase (MAPK) signaling pathways, as well as the PI3K-Akt and Ras signaling pathways. We speculate that <i>METTL3</i> may increase the level of m6A methylation to inhibit fat deposition and/or inhibit the expression of HSP genes to enhance the body’s resistance to heat stress, while the <i>FTO</i> gene generated the opposite molecular mechanism. This study provides a scientific basis and theoretical support for sheep feeding and management practices during heat stress. |
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| ISSN: | 1661-6596 1422-0067 |