ABSTRACT: RNA sequencing was performed to obtain a landscape of mRNA, circRNA, lncRNA, and miRNA of bovine intramuscular adipocytes in 4 differentiation periods.
Project description:To find the differentially and highly expressed circRNAs between the early and late stages of intramuscular adipocytes differentiation. Explored their expression profiles and further predicted their possible interaction relationships. To generated the final circRNA-miRNA interaction network that have valuable functions in adipocyte differentiation.
Project description:To investigate the function circPPARG1 in the regulation of adipogenic differentiation, we overexpressed circPPARG1 in Intramuscular adipocytes
Project description:To investigate the function lncFABP4 in the regulation of adipogenic differentiation, we overexpressed lncFABP4 in buffalo intramuscular adipocytes
Project description:Abstract Background: Intramuscular fat (IMF) and subcutaneous fat (SF) are two major adipose depots in cattle, each playing distinct roles in meat quality and systemic metabolism. The development of adipose tissue requires proliferation and differentiation of preadipocytes into adipocytes. However, the identity and characteristics of preadipocytes forming IMF and SF, especially the former, remain poorly defined. Results: In this study, we derived four single-preadipocyte clones from stromal vascular fraction (SVF) of IMF and SF tissues of four adult steers and characterized their gene expression signatures. All single-cell clones of IMF and SF preadipocytes were capable of forming adipocytes, as evidenced by lipid droplets accumulation and upregulation of adipocyte marker genes such as PPARG, CEBPA, FABP4, and ADIPOQ upon adipogenic induction. Transcriptomic analysis revealed that although both IMF and SF preadipocyte clones expressed classical preadipocyte markers such as PDGFRA, DLK1, PPARG, CD34, and ZNF423, they had significantly different gene expression profiles. Over 2,000 genes were differentially expressed between IMF and SF preadipocyte clones. Surprisingly, many muscle-specific genes such as MYOG, CKM, MYH2, and MYH3 were expressed at high levels in IMF preadipocyte clones while barely detectable in SF preadipocyte clones. These muscle-specific genes remained elevated in adipocytes differentiated from IMF preadipocyte clones. Functional enrichment analysis revealed that genes upregulated in IMF preadipocyte clones compared to SF preadipocyte clones were significantly enriched in pathways such as PI3K-Akt signaling, MAPK signaling, calcium signaling, cholesterol biosynthesis, muscle structure development, and neuromuscular junction formation, further suggesting that IMF preadipocytes bear characteristics of muscle cells. Differentiation assays demonstrated that IMF SVF cells had strong adipogenic potential, as they readily formed adipocytes upon adipogenic induction, but showed little myogenic potential, as they did not form myotubes upon myogenic induction. In contrast, bovine satellite cells, the widely considered myogenic progenitor cells in skeletal muscle, exhibited both myogenic and adipogenic potential, forming both myotubes and adipocytes under respective induction conditions. Conclusions: These results demonstrate that IMF and SF preadipocytes differ significantly in gene expression profiles with the former expressing many muscle-specific genes. Our findings suggest that IMF preadipocytes may originate from muscle satellite cells or share a common progenitor, whereas SF preadipocytes arise from non-myogenic adipose progenitor cells.
Project description:Beef marbling is caused by intramuscular deposition, and it is an economically important trait in the beef industry. Vitamin A (VA) is an important feed supplement for cattle, but it can hinder marbling if provided in excess. In cattle, VA forms various derivatives such as all-trans retinoic acid (ATRA) and 9-cis retinoic acid (9cRA). Therefore, we investigated the genes involved in bovine intramuscular adipogenesis after VA treatment with ATRA and 9cRA. Differential gene expression levels were validated by microarray analysis in a clonal bovine intramuscular preadipocyte (BIP) cell line derived from the intramuscular adipose tissue of Japanese Black cattle. BIP cells were harvested six days after adipogenic stimulation with either 1 μM ATRA, 1 μM 9cRA, or nonretinoic acids control. The ATRA- and 9cRA-treated cells exhibited reduced transcription of genes involved in the circulatory system and muscle development compared with the no retinoic acid (RA) treatment. In addition, the ATRA- and 9cRA-treated cells exhibited increased transcription of genes involved in the immune system, protein kinase B signaling, and responses to various stimuli. These results demonstrate the lower expression of muscle development in ATRA- and 9cRA-treated BIP cells during adipogenesis.
Project description:Beef marbling is caused by intramuscular deposition, and it is an economically important trait in the beef industry. Vitamin A (VA) is an important feed supplement for cattle, but it can hinder marbling if provided in excess. In cattle, VA forms various derivatives such as all-trans retinoic acid (ATRA) and 9-cis retinoic acid (9cRA). Therefore, we investigated the genes involved in bovine intramuscular adipogenesis after VA treatment with ATRA and 9cRA. Differential gene expression levels were validated by microarray analysis in a clonal bovine intramuscular preadipocyte (BIP) cell line derived from the intramuscular adipose tissue of Japanese Black cattle. BIP cells were harvested six days after adipogenic stimulation with either 1 ?M ATRA, 1 ?M 9cRA, or nonretinoic acids control. The ATRA- and 9cRA-treated cells exhibited reduced transcription of genes involved in the circulatory system and muscle development compared with the no retinoic acid (RA) treatment. In addition, the ATRA- and 9cRA-treated cells exhibited increased transcription of genes involved in the immune system, protein kinase B signaling, and responses to various stimuli. These results demonstrate the lower expression of muscle development in ATRA- and 9cRA-treated BIP cells during adipogenesis. BIP cells were cultured according to previously reported methods (Aso et al. 1995, Mizoguchi et al. 2014). Confluent cultures were transferred to fresh Dulbecco’s modified Eagle’s medium, which contained 50 ng/mL insulin, 0.25 ?M dexamethasone, 5 mM octanoate, 10 mM acetic acid, 10% fetal bovine serum, 100 U/mL penicillin, and 100 ?g/mL streptomycin. The cells were cultured for up to 6 days and the medium was changed every 2 days. BIP cells were treated with ATRA (1 ?M), 9cRA (1 ?M), or they received no treatment (control).