Effect of<i>INSIG1</i>on the milk fat synthesis of buffalo mammary epithelial cells

Fan, Xinyang; Qiu, Lihua; Teng, Xiaohong; Zhang, Yongyun; Miao, Yongwang

doi:10.1017/s0022029920000710

Cited by 20 publications

(15 citation statements)

References 39 publications

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“…The BuMECs were obtained from five buffalo in lactation (60 days postpartum) as previously reported by our group (Fan et al, 2020). Briefly, the buffalo mammary tissues were stripped of connective tissues.…”

Section: Methodsmentioning

confidence: 99%

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Elongase of very long chain fatty acids 6 (ELOVL6) promotes lipid synthesis in buffalo mammary epithelial cells

Fan

Zhu

Qiu

et al. 2021

Animal Physiology Nutrition

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Recent studies have shown elongase of very‐long‐chain fatty acids 6 (ELOVL6) is a vital protein for endogenous synthesis of saturated and monounsaturated long‐chain fatty acids in some mammals. Nevertheless, its role in lipid synthesis in buffalo mammary gland is still unclear. In this work, the full‐length coding sequence (CDS) of ELOVL6 was cloned and identified from buffalo mammary gland. As a result, the CDS of this gene is 795 bp, which encodes a polypeptide of 264 amino acid residues. The buffalo ELOVL6 contains an ELO domain which belongs to the ELO superfamily. Among the 10 tissues of buffalo in peak lactation detected by RT‐qPCR, the expression level of ELOVL6 was the highest in the brain, followed by the spleen, and then decreased in the mammary gland, muscle, kidney, heart, liver, rumen, intestine and lung. However, only the expression in the brain and spleen was statistically different from that in other tissues (p < 0.05). Compared with that of the dry‐off period, the mRNA abundance of ELOVL6 in the mammary gland was significantly increased in peak lactation. The experiments based on lentivirus transfection in buffalo mammary epithelial cells (BuMECs) displayed that the overexpression of ELOVL6 markedly promoted the expression of INSIG1, INSIG2, SREBP, PPARG, FASN, GPAM, DGAT2 and APGAT6 genes, and the knockdown of ELOVL6 significantly decreased the mRNA abundance of INSIG2, SREBP, FASN, SCD, GPAM, APGAT6 and TIP47 genes. In addition, the increase or decrease of ELOVL6 expression level also caused the corresponding change of total triglyceride content in the BuMECs. The results here suggest that the ELOVL6 can catalyse the synthesis of long‐chain fatty acids in the BuMECs, and it can indirectly affect the expression of genes related to milk fat synthesis through its catalytic products to promote the lipid biosynthesis of BuMECs.

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Section: Methodsmentioning

confidence: 99%

“…The process for generation and transduction was performed as previously reported (Fan et al, 2020). Briefly, the 239 T cells were, respectively, transfected with pLVX‐ELOVL6‐IRES‐ZsGreen1 and pLVX‐IRES‐ZsGreen1 vector (Lv‐GFP) for packaging lentiviral particles.…”

Section: Methodsmentioning

confidence: 99%

Elongase of very long chain fatty acids 6 (ELOVL6) promotes lipid synthesis in buffalo mammary epithelial cells

Fan

Zhu

Qiu

et al. 2021

Animal Physiology Nutrition

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“…Insulin inducible genes INSIG1 and INSIG2 are located in endoplasmic reticulum and play an important role in regulating lipid metabolism ( 75 ). The mRNA abundance of INSIG1 in breast tissue during peak lactation period was lower than that in dry period, which reveal the potential importance of INSIG in milk fat metabolism ( 76 ). After overexpression INSIG1 or INSIG2 in GMECs, the expression of ACACA and FASN was down-regulated, the content of TAG and total cholesterol (TC) were decreased.…”

Section: Negative Regulation Of De Novo Synthesis Of Famentioning

confidence: 99%

“…Previous studies have shown that INSIG1 can effectively block the proteolysis activation of SREBP family members ( 114 ) and play an important role in de novo milk fat synthesis in ruminants ( 50 ). Fan et al ( 76 ) measured the protein abundance of INSIG1 in the mammary gland of buffalo during peak lactation and dry period, and found that the expression of INSIG1 in the peak lactation period was lower than that in the dry period, and the overexpression INSIG1 significantly decreased the expression of GPAM and AGPAT6. The opposite result was obtained after INSIG1 was knockout, which was also confirmed by Li et al ( 77 ).…”

Section: Tag Synthesismentioning

confidence: 99%

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Regulation of Key Genes for Milk Fat Synthesis in Ruminants

et al. 2021

Front. Nutr.

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Milk fat is the most important and energy-rich substance in milk and plays an important role in the metabolism of nutrients during human growth and development. It is mainly used in the production of butter and yogurt. Milk fat not only affects the flavor and nutritional value of milk, but also is the main target trait of ruminant breeding. There are many key genes involve in ruminant milk fat synthesis, including ACSS2, FASN, ACACA, CD36, ACSL, SLC27A, FABP3, SCD, GPAM, AGPAT, LPIN, DGAT1, PLIN2, XDH, and BTN1A1. Taking the de novo synthesis of fatty acids (FA) and intaking of long-chain fatty acids (LCFA) in blood to the end of lipid droplet secretion as the mainline, this manuscript elucidates the complex regulation model of key genes in mammary epithelial cells (MECs) in ruminant milk fat synthesis, and constructs the whole regulatory network of milk fat synthesis, to provide valuable theoretical basis and research ideas for the study of milk fat regulation mechanism of ruminants.

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Circular RNA_015343 sponges microRNA‐25 to regulate viability, proliferation, and milk fat synthesis of ovine mammary epithelial cells via INSIG1

Wu,

Wang,

Hao

et al. 2024

Journal Cellular Physiology

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In our previous study, circ_015343 was found to inhibit the viability and proliferation of ovine mammary epithelial cells (OMECs) and the expression levels of milk fat synthesis marker genes, but the regulatory mechanism underlying the processes is still unclear. Accordingly in this study, the target relationships between circ_015343 with miR‐25 and between miR‐25 with insulin induced gene 1 (INSIG1) were verified, and the functions of miR‐25 and INSIG1 were investigated in OMECs. The dual‐luciferase reporter assay revealed that miR‐25 mimic remarkably decreased the luciferase activity of circ_015343 in HEK293T cells cotransfected with a wild‐type vector, while it did not change the activity of circ_015343 in HEK293T cells cotransfected with a mutant vector. These suggest that cic_015343 can adsorb and bind miR‐25. The miR‐25 increased the viability and proliferation of OMECs, and the content of triglycerides in OMECs. In addition, INSIG1 was found to be a target gene of miR‐25 using a dual‐luciferase reporter assay. Overexpression of INSIG1 decreased the viability, proliferation, and level of triglycerides of OMECs. In contrast, the inhibition of INSIG1 in expression had the opposite effect on activities and triglycerides of OMECs with overexpressed INSIG1. A rescue experiment revealed that circ_015343 alleviated the inhibitory effect of miR‐25 on the mRNA and protein abundance of INSIG1. These results indicate that circ_015343 sponges miR‐25 to inhibit the activities and content of triglycerides of OMECs by upregulating the expression of INSIG1 in OMECs. This study provided new insights for understanding the genetic molecular mechanism of lactation traits in sheep.

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Effect ofINSIG1on the milk fat synthesis of buffalo mammary epithelial cells

Cited by 20 publications

References 39 publications

Elongase of very long chain fatty acids 6 (ELOVL6) promotes lipid synthesis in buffalo mammary epithelial cells

Elongase of very long chain fatty acids 6 (ELOVL6) promotes lipid synthesis in buffalo mammary epithelial cells

Regulation of Key Genes for Milk Fat Synthesis in Ruminants

Circular RNA_015343 sponges microRNA‐25 to regulate viability, proliferation, and milk fat synthesis of ovine mammary epithelial cells via INSIG1

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