ABSTRACT
Skin is the body's largest organ, and the main function of skin is to protect underlying organs from possible external damage. Melanocytes play an important role in skin pigmentation. The Bama pig has a "two-end-black" phenotype with different coat colors across skin regions, e.g., white skin (without melanocytes) and black skin (with melanocytes), which could be a model to investigate skin-related disorders, specifically loss of melanocytes. Here, we generated expression profiles of mRNAs and long noncoding RNAs in Bama pig skins with different coat colors. In total, 14,900 mRNAs and 7549 lncRNAs were expressed. Overall, 2338 mRNAs/113 lncRNAs with FDR-adjusted p-value ≤ 0.05 were considered to be differentially expressed (DE) mRNAs/lncRNAs, with 1305 down-regulated mRNAs and 1033 up-regulated mRNAs in white skin with|log2(fold change)| > 1. The genes down-regulated in white skin were associated with pigmentation, melanocyte-keratinocyte interaction, and keratin, while up-regulated ones were mainly associated with cellular energy metabolisms. Furthermore, those DE lncRNAs were predicted to be implicated in pigmentation, keratin synthesis and cellular energy metabolism. In general, this study provides insight into the transcriptional difference involved in melanocyte-loss-induced keratinocyte changes and promotes the Bama pig as a biomedical model in skin research.
Subject(s)
Gene Expression Profiling/methods , Melanocytes/cytology , RNA, Long Noncoding/genetics , RNA, Messenger/genetics , Skin/chemistry , Animals , Energy Metabolism , Gene Expression Regulation , Melanocytes/chemistry , Phenotype , Principal Component Analysis , Sequence Analysis, RNA , Skin/cytology , Skin Pigmentation , SwineABSTRACT
Natural antisense transcripts (NATs) are widely present in mammalian genomes and act as pivotal regulator molecules to control gene expression. However, studies on the NATs of pigs are relatively rare. Here, we identified a novel antisense transcript, designated PLA2G16-AS, transcribed from the phospholipase A2 group XVI locus (PLA2G16) in the porcine genome, which is a well-known regulatory molecule of fat deposition. PLA2G16-AS and PLA2G16 were dominantly expressed in porcine adipose tissue, and were differentially expressed between Tibetan pigs and Rongchang pigs. In addition, PLA2G16-AS has a weak sequence conservation among different vertebrates. PLA2G16-AS was also shown to form an RNA-RNA duplex with PLA2G16, and to regulate PLA2G16 expression at the mRNA level. Moreover, the overexpression of PLA2G16-AS increased the stability of PLA2G16 mRNA in porcine cells. We envision that our findings of a NAT for a regulatory gene associated with lipolysis might further our understanding of the molecular regulation of fat deposition.
Subject(s)
Adipose Tissue/metabolism , Phospholipases A2, Calcium-Independent/genetics , RNA, Long Noncoding/genetics , RNA, Messenger/chemistry , Animals , Cell Line , Evolution, Molecular , Gene Expression Regulation , Organ Specificity , Phospholipases A2, Calcium-Independent/chemistry , RNA Stability , RNA, Long Noncoding/metabolism , Species Specificity , Sus scrofa , Tissue DistributionABSTRACT
OBJECTIVE: To find out the optimal extraction technique for HDCA (a-hydroxycholic acid). METHOD: According to the orthogonal design L9(3(4)), the optimal extraction technique was sought through experimental investigation, and the content of HDCA was determined by TLC. RESULT: The optimal extraction method was eightfold 8% NaOH solution and 16 hours. The optimal purification method was six fold ethyl acetate, 5% active carbon, and 30 minutes twice. CONCLUSION: The above mentioned extraction technique is optimal and feasible in extraction of HDCA.
