ABSTRACT
miR-101a promotes the differentiation of goat skeletal muscle satellite cells (SMSCs), as we previously reported, but the underpinning mechanism remains to be illuminated. In this study, we predicted the target gene of miR-101a by employing online softwares PicTar, TargetScan and miRanda, and found that enhancer of zeste homologue 2 (EZH2) was targeted by miR-101a. Further we identified that EZH2 contained miR-101a binding sites at its 3'UTR by using the dual-luciferase reporter assay system. In addition, we showed that during SMSC differentiation, the downregulated levels of EZH2 mRNA and protein were accompanied by increasing miR-101a expression via qRT-PCR and Western blot. Additionally, the expression of EZH2 significantly increased (P<0.01) when miR-101a was suppressed, whereas overexpressing miR-101a almost had no effect on EZH2 expression (P>0.05). These data demonstrated that miR-101a promotes SMSC differentiation directly through EZH2, which provides a theoretical reference for further elucidating the mechanism of miR-101a in SMSC differentiation.
Subject(s)
Cell Differentiation/genetics , Enhancer of Zeste Homolog 2 Protein/genetics , MicroRNAs/genetics , Satellite Cells, Skeletal Muscle/physiology , 3' Untranslated Regions/genetics , Amino Acid Sequence , Animals , Binding Sites/genetics , Cell Line , Down-Regulation/genetics , GoatsABSTRACT
Postmortem examinations were made in 99 goats in Nimu County of Tibe, and parasites were collected and identified based on morphology. The collected parasites were categorized, and infection status was analyzed. The helminth infection rate was 100% among the goats, and all showed a pattern of mixed infection. The identified parasites belonged to 21 species, 15 genera, and 9 families. The Trichuris genusï¼36.4%ï¼ was the most prevailing among nematodes in the gastrointestinal tract; Paramphistomum cerviï¼60.6%ï¼ and Paramphistomum gotoiï¼60.6%ï¼ were predominant among trematodes detected; Cysticercus tenuicollisï¼52.5%ï¼ was the predominant cestode detected; and Orientobilharzia turkestanicum was the major parasite detected in the portal vein (69.7%).
Subject(s)
Goat Diseases , Helminthiasis, Animal , Animals , Gastrointestinal Tract , Goats , Helminthiasis , Nematoda , TibetABSTRACT
Integrin linked kinase (ILK) is a scaffold protein, which plays important roles in hair follicle development. The cDNA sequence of novel ILK gene in sheep was cloned by PCR method and analyzed by bioinformatics. Tissue expression profiling in eight tissues and temporal profiling at different wool follicle anagen stages in skin was analyzed. The results showed that the whole open reading frame (ORF) of ILK gene was 1 359 bp in length, which encoded 452 amino acids. Bioinformatic analysis indicated that the secondary structure of ILK gene was mainly made up of three ankyrin repeats and a kinase domain, and there were multiple phosphorylation and Protein Kinase C sites in this gene. The RT-PCR result confirmed that ILK mRNA was expressed in heart, liver, spleen, lung, skeletal muscle, skin, and small intestine, and the expression level was much higher in skin, spleen, and liver than others. The q-PCR analysis demonstrated that the ex-pression level of ILK was significantly increased from March to May (early follicle anagen initiation) in both sheep breeds, Chinese Merino and Kazakh sheep, and there were certain differences from June to October between the two breeds. The above results indicated that ILK gene may play key roles in regulating secondary follicle growth.
