Hoxa5 Inhibits the Proliferation and Induces Adipogenic Differentiation of Subcutaneous Preadipocytes in Goats

Abstract

The homeobox a5 (Hoxa5) plays considerable roles in the differentiation and lipid metabolism of adipocytes. However, the current knowledge about the mechanistic roles and functions of Hoxa5 in goat subcutaneous preadipocyte remains unclear. Therefore, Hoxa5 loss-of-function and gain-of-function was performed to reveal its functions in adipogenesis. For differentiation, overexpression of Hoxa5 notably increased the expression of adipogenic genes (PPARγ, CEBP/α, CEBP/β, AP2, and SREBP1), as well as promoted goat subcutaneous preadipocyte lipid accumulation. Knockdown of Hoxa5 mediated by siRNA technique significantly inhibited its differentiation and suppressed the accumulation of lipid droplets. Regarding proliferation, overexpression of Hoxa5 reduced the number of cells stained with crystal violet, and inhibited mRNA expression of the marker genes including CCNE1, PCNA, CCND1, and CDK2, and also significantly reduced EdU-positive rates. Consistently, knockdown of Hoxa5 demonstrated the opposite tendency. In conclusion, these data demonstrated that Hoxa5 promotes adipogenic differentiation of goat subcutaneous preadipocyte and inhibits its proliferation in vitro.

miR-130b duplex (miR-130b-3p/miR-130b-5p) negatively regulates goat intramuscular preadipocyte lipid droplets accumulation by inhibiting Krüppel-like factor 3 expression

“…The same result was observed in the rescue experiment of this study. Also, the same problem has been found in previous studies ( Guo et al, 2020 ; Xu et al, 2021 ; Chen et al, 2022 ). In addition, pref-1 , a well-known negative regulator of adipogenesis, was positively correlated with miR130b duplex (miR-130b-3p and miR-130b-5p) and KLF3 regulated goat intramuscular adipocyte differentiation.…”

Effects of HOXC8 on the Proliferation and Differentiation of Porcine Preadipocytes

“…The epigenetic dysregulation of the HOXA5 gene is associated with adipocyte hypertrophy in human obesity [ 38 ] and contributes to adipose differentiation in mice [ 39 , 40 ]. Consistent results for HOXA5 in adipogenesis were obtained in goats [ 41 ]. HOXC10 negatively regulates lipid droplet accumulation by suppressing the expression of lipoprotein lipase in sheep bone marrow mesenchymal stem cells [ 13 ].…”

FOS Inhibits the Differentiation of Intramuscular Adipocytes in Goats

“…Jianzhou Daer goats (Capra hircus) (N 3) were purchased from Sichuan Tiandi Goat Biological Engineering Co., Ltd. (Chengdu, China). Goat intramuscular preadipocytes were isolated and cultured in accordance with previously described methods [15,16]. Concisely, Longissimus dorsi muscle samples were collected from seven-day-old Jianzhou Daer goats (N = 3), and the samples were sheared.…”

“…Bodipy and DAPI stainings were performed according to the method described by Chen et al [16]. Bodipy and DAPI dyes were first diluted in PBS at a ratio of 1:1000.…”

“…The A260/A280 values of the RNA were determined as in the range 1.8-2.0 using a UV spectrophotometer, which met the test requirements. cDNA was reverse-transcribed using a reverse-transcriptase kit (Thermo, Waltham, MA, USA) and the quantitative PCR spiking system described by Chen et al [16]. The UXT gene was used as an internal control.…”