Mongolian Horse Transcriptome And MiRNA Library Characteristics And Correlation Analysis Before And After High Load Exercise Training

The Mongolia horse is a typical grassland breed, in the long process of natural selection and artificial selection, they’ve been survived from hard nature condition, so the Mongolian horses’ genome contains a large number of genes, which had priceless value,such as: stamina, resistance, etc. This research took 6 Mongolian horses for 4 months high load training, collected blood and muscle samples before and after the training respectively, detected the change of physiological and biochemical indexes by blood,using the next generation sequencing technology to establish the transcriptome and mi RNA library by muscle samples at the same time, enriched the differentially expressed genes to the GO analysis and KEGG pathway, found the significant different metabolic pathways and related genes. Later, we did correlation analysis for transcriptome results and mi RNA results. getting the regulation network diagram of mi RNA-m RNA before and after the training. We established Pre-mi R-1 expression vector and successfully transfected into Mongolian horse in skeletal muscle cells by in vitro culture. verified the mi RNA-m RNA targeted regulation relationship.The main results were as follows:1. after the training, GLU, CK, MCH, MCHC, PLT, PCT increase significantly; and significant drop of UR, HGB, RBC, WBC, Lymph, Mon, Gran, RDW; no significant change found in MPV, HCT, MCV, LDH, AST, ALT.2. Screened out 1102 different express genes in transcriptome library, 299 were up regulated, and 803 were down regulated. These DEGs were annotation to 3398 GO terms, including muscle structure development, cardiovascular system development,cardiovascular system, circulatory system development, muscle tissue development,muscular organ development, muscle contraction, muscle cell differentiation, etc. and they were annotated to 193 KEGG pathways, athletics related pathway including:Dilated cardiomyopathy(DCM), Hypertrophic cardiomyopathy, Cardiac muscle contraction pathway, Calcium signaling pathway, the actin cytoskeleton adjustment pathway, etc.3. we got 248 different express mi RNAs in library, 54 were up regulated and 194 were down regulated. the GO enrichment results showed that the mi RNA’s target genes were annotated to developmental process, anatomical structure development, protein binding, enzyme binding, etc. In KEGG analysis, 531 target genes were annotated to210 pathways, the significantly enriched pathway includes: actin cytoskeleton metabolic, Calcium signaling pathways, Cardiac muscle contraction, etc.4. combining the transcriptome and mi RNA library results, we found 29 mi RNA targeted on 236 genes. 15 mi RNA down regulated 68 genes and 14 mi RNA up regulated 168 genes. The GO enrichment results showed the significant target genes were annotated to system development, anatomical structure development etc. the KEGG enrichment results showed 85 pathways such as: regulation of actin cytoskeleton, calcium signaling pathways, myocardial contraction and hypertrophic cardiomyopathy, dilated cardiomyopathy, etc. were significantly annotated.5. Establishing Pre-mi R-1 expression vector and transfected into Mongolian horse in skeletal muscle cells. After the relative expression detection of mi RNA-1, Pre-mir-1and IGF1 by RT-PCR, We came to a conclusion that the Pre-mir-1 transfected into skeletal muscle cells successfully, processing into mature mi RNA-1, then inhibit or silence the target gens’ s expression, to verify the targeted regulation relationship between mi RNA-1 and IGF1, also demonstrate the validity of the m RNA-mi RNA correlation analysis results at the same time.