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

TL;DR

This study uses nanopore sequencing to explore how gene expression changes in pig testes before and after sexual maturity, revealing key regulatory networks involved in reproduction.

Contribution

The study provides a comprehensive analysis of full-length transcripts and non-coding RNAs in porcine testes during sexual maturation using long-read sequencing.

Findings

11,060 mRNAs, 15,338 transcripts, 688 lncRNAs, and 19 circRNAs were differentially expressed between pre- and post-sexual maturity testes.

Differential RNAs were linked to spermatogenesis, metabolic pathways, and Wnt/MAPK signaling, with core genes like PRM1 and ODF2 showing synergistic expression.

DELs and DEcircRNAs were associated with steroid biosynthesis and spermatogenesis, offering potential molecular markers for reproductive efficiency.

Abstract

Background: 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. Methods: 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. Results: 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., PRM1, ODF2, GSTM3) 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. Conclusions: 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.

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