Nanopore Sequencing Technology Reveals the Transcriptional Expression Characteristics of Male Pig’s Testes Before and After Sexual Maturity

<b>Background</b>: Testicular development and spermatogenesis are intricate biological processes controlled by a coordinated transcriptional network. However, comprehensive characterization of full-length transcripts and non-coding RNAs (ncRNAs) during porcine testicular sexual maturation remains limited. <b>Methods</b>: This study systematically profiled the transcriptional landscape of pig testes prior to (pre-sexual maturity, PSM) and following (post-sexual maturity, SM) sexual maturity using Oxford Nanopore Technologies (ONT) long-read sequencing. <b>Results</b>: There were 11,060 differentially expressed mRNAs (DEGs), 15,338 differentially expressed transcripts (DETs), 688 differentially expressed lncRNAs (DELs), and 19 differentially expressed circRNAs (DEcircRNAs) between PSM and SM groups among the 9941 mRNAs, 15,339 transcripts, 4136 lncRNAs (58.58% being LincRNAs). These differential RNAs converged on 133 shared GO terms (e.g., spermatogenesis, male gamete generation) and 58 common KEGG pathways (e.g., metabolic pathways, Wnt/MAPK signaling), according to functional enrichment and combined analysis. Core genes (e.g., <i>PRM1</i>, <i>ODF2</i>, <i>GSTM3</i>) demonstrated synergistic expression across gene, transcript, lncRNA-cistarget, and circRNA levels. Furthermore, DELs were associated with steroid biosynthesis and N-glycan biosynthesis, whereas DEcircRNAs, which were mostly upregulated after puberty, were thought to control genes linked to spermatogenesis. <b>Conclusions</b>: This research sheds light on the dynamic transcriptional reprogramming that occurs during the maturation of pig testicles, advances our knowledge of coding and ncRNA regulatory networks in male mammals, and offers useful molecular markers for enhancing pig reproductive efficiency.