Abstract: In order to identify and analyze alternative splicing events（AS） in skeletal muscle of Mongolian cattle during cold resistance, the Illumina Novaseq^TM 6000 platform was used for transcriptomic sequencing of skeletal muscle tissues of Mongolian cattle in winter and summer, respectively. The AS event and differential splicing gene（DSG） between winter and summer in skeletal muscle of Mongolian cattle were identified using rMATS software. The GO function enrichment and KEGG pathway analysis were performed for DSGs. The results showed that a total of 272 950 322 raw reads were obtained by transcriptomic sequencing, and there were 10 966 and 13 756 AS events in the winter and summer groups, respectively. Skipped exon（SE） accounted for 78.64% and 82.48% of AS events in winter and summer groups, respectively. A total of 843 significant DSGs were identified between winter and summer groups. DSGs enriched 380, 105 and 174 GO terms in biological processes, cell components and molecular functional categories, respectively. DSGs were enriched in 20 signal pathways, among which PPAR signal pathway, insulin resistance, TCA cycle and circadian rhythm pathways which were closely related to energy metabolism were significantly enriched. The results indicated that there might be significant differences in lipid metabolism, oxidative stress, insulin sensitivity and circadian rhythm in Mongolian cattle skeletal muscle between winter and summer.