Isolation, characterization, and SREBP1 functional analysis of mammary epithelial cell inbuffalo

Abstract

Compared to cow milk, buffalo milk contains more protein, fat, and vitamin. Buffalo milk is an ideal food in human life. Sterol regulatory element‐binding protein 1 (SREBP1), an important transcription factor, regulates the expression and activity of enzyme and protein involved in milk fat synthesis to influence on the synthesis and secretion of triglyceride in mammary epithelial cells. In the present study, we successfully isolated buffalo mammary epithelial cell by using enzymatic digestion, and then described the growth characteristics and expression characteristics of mammary epithelial cells. Moreover, we cloned the SREBP1 gene from total RNA isolated from milk fat globule and analyzed the function of the SREBP1 gene. After infected with shRNA‐SREBP1 lentiviral particle and treated with fatty acid, the expression trend of ACACA, FABP3, FAS, SCD, ERK1, ERK2, PPARy, and Insigl genes was consistent with the expression trend of SREBP1 gene. These results suggested that SREBP1 gene is a central transcription factor in regulating milk fat synthesis and SREBP1 gene may act on ERK1/ERK2 signaling pathway to regulate the expression of PPARy gene. The current study will provide a theoretical basis for further reveal the molecular mechanism of milk fat synthesis in buffalo mammary epithelial cells. Practical applications This study aim to separate and analysis characterization of mammary epithelial cell in buffalo. Compared to cow milk, buffalo milk contains more protein, fat, and vitamin. Buffalo milk is an ideal food in human life. This study will provide a theoretical basis for further research on the molecular mechanism of milk fat synthesis in buffalo mammary epithelial cells.

MicroRNA-212 targets SIRT2 to influence lipogenesis in bovine mammary epithelial cell line

“…It is generally accepted that PPAR γ is a critical factor for lipogenesis and has relationships with other lipogenic genes (Farmer, 2006; Rosen and MacDougald, 2006; Siersbaek et al ., 2012). In buffalo, SREBP1 gene may act on ERK1 / ERK2 signaling pathway to regulate the expression of PPARy gene (Xu et al ., 2019 a , 2019 b ) and, as a transcription regulator, PPAR γ is able to enhance the expression of many genes related to lipogenesis including FASN (Lefterova et al ., 2008; Zhang et al ., 2016). Moreover, many other studies have indicated that these genes (PPAR γ, FASN , SREBP1 ) are associated with milk fat synthesis in mammary epithelial cells (Yang et al ., 2017).…”

“…Many studies have confirmed that SIRT2 is involved in lipogenesis (Perrini et al ., 2020; Xu et al ., 2019 a , 2019 b ; Lantier et al ., 2018) and the co-transfection experiment showed that by targeting SIRT2 , miR-212 upregulates the mRNA expression of FASN and SREBP1 but not PPAR γ. These are the first data to confirm that SIRT2 can directly regulate FASN and SREBP1 negatively, whilst the result of PPAR γ is at odds with a recent report (Xu et al ., 2019 a , 2019 b ) that in bovine ovarian granular cells, SIRT2 knockdown or treatment with inhibitors produced negative effects on PPAR γ. It was supposed that the mechanism might be SIRT2 mediating the transcription factors FOXO1 .…”

The 84-bp Indel Polymorphism of The Sterol Regulatory Element-Binding Protein 1 (SREBP1) Gene in Several Cattle Breeds in Indonesia

“…SREBP1 or also known as sterol regulatory element binding transcription factor 1 (SREBF1) is also essential in the fat metabolism of ruminants because it also influences the fat characteristics of both meat and dairy products. This has been demonstrated in cattle (Harvatine & Bauman 2006;Proskura et al 2017;Gamarra et al 2021;Kawaguchi et al 2021), buffalo (Deng et al 2017;Xu et al 2019), goat (Xu et al 2016), and sheep (Tsiplakou et al 2015;Liang et al 2020;Carcangiu et al 2021). In bovine, the SREBP1 gene is located on chromosome 19 with 21 exons and encodes 1183 amino acids (Hoashi et al 2007).…”