Seminar Title
Analysis of RNA Population in Goat Spermatozoa
Seminar Type
Defence Seminar
Biotechnology and Medical Engineering
Speaker Name
Bijayalaxmi Sahoo ( Rollno : 515bm1001)
Speaker Type
Hybrid Mode: BM Seminar Room + Online in MS Teams (URL: Code: 9j41igc
Date & Time
28 Sep 2023 11:30 AM
Prof. Mukesh Kumar Gupta

This study investigated the RNA population in goat spermatozoa by RLM-RACE, long-read Nanopore sequencing, and short-read Illumina sequencing. The effects of heparin- and arginine-induced sperm capacitation and motility enhancement on spRNAs were also investigated. Results showed that goat spermatozoa contain 20.03 fg of RNA, which included mRNA, rRNA, miscRNA, miRNA, circRNA, etc. Nanopore sequencing resulted in long-sized reads of 500bp to 2Kb, which could be mapped to 123 transcripts by de novo assembly. On the other hand, full-length cDNA cloning by RLM-RACE identified 31 transcripts. The full-length transcripts of five fertility-related genes (AR, CATSPER3, DAZL, HSP90AA1, SPACA1, PRM3) were found in the goat spermatozoa. The short-read RNA-seq Illumina discovered 16604 transcripts, including 357 mRNAs, 127 circRNAs, 655 lncRNAs, and 160 imprinted genes. The coding mRNAs were involved in different biological pathways, viz. Wnt signaling, cAMP-signaling, AMPK signaling, and MAPK signaling pathways. Results further showed that heparin-induced capacitation upregulated 1254 transcripts, including genes (PRND, LLCFC1, SPESP1) that were previously reported to participate in signaling pathways of sperm capacitation and acrosome reaction. Similarly, arginine-induced sperm capacitation and motility enhancement were also associated with the upregulation of 1321 genes. The differentially expressed genes in both the heparin- and arginine-treated groups were involved in cAMP/PKA signaling, PI3-Akt signaling, MAPK signaling, calcium signaling, and oxidative stress pathways. DCFDA staining confirmed the increased ROS generation in goat spermatozoa upon treatment with heparin or arginine. In conclusion, this study documents the suite of RNA populations in goat spermatozoa and their relevance in heparin- and arginine-induced sperm capacitation. A molecular mechanism of heparin- and arginine-mediated sperm capacitation is proposed.