To investigate the biological role of the lncRNA NEAT1 in porcine skeletal muscle development, this study employed a series of molecular and cellular approaches. First, the full-length cDNA sequence of porcine NEAT1 was determined using RACE technology, revealing a 3284-base pair transcript. RT-qPCR was then used to analyze the spatiotemporal expression patterns of NEAT1 in various porcine tissues and PSCs at different developmental stages, while its subcellular localization was characterized. Finally, siRNA-mediated silencing of NEAT1 was performed to assess its effects on PSCs proliferation and differentiation. RT-qPCR analysis showed that NEAT1 is highly expressed in porcine heart and skeletal muscle tissues. During PSCs development, NEAT1 expression is dynamically regulated: it is significantly upregulated during the proliferation phase and markedly downregulated upon entry into the differentiation phase. Subcellular localization studies demonstrated that NEAT1 is distributed in both the cytoplasm and nucleus of PSCs, with predominant ctoplasmic localization during proliferation; as differentiation proceeds, cytoplasmic NEAT1 abundance decreases, resulting in distinct subcellular expression patterns between the two stages. Functional validation revealed that silencing NEAT1 significantly reduces PSC proliferative activity, accompanied by downregulation of key proliferation markers (PCNA, CCNA2, CCNB1, CCNE2, CCND1, and CDK4; P < 0.05) and upregulation of the cell cycle inhibitor CDKN1A (P < 0.05). Concurrently, NEAT1 silencing enhances PSC differentiation, as evidenced by increased expression of muscle differentiation markers (MyoD, MyoG, and MyHC; P < 0.05). I conclusion, NEAT1 plays a critical role in porcine skeletal muscle development by promoting PSCs proliferation and inhibiting their differentiation. These findings provide a foundation for further exploration of the molecular regulatory mechanisms underlying NEAT1 function in porcine skeletal muscle development.

Highlights

• This study is the first to systematically characterize the full-length cDNA sequence of porcine NEAT1 (3284 bp) and clarify its dynamic spatiotemporal expression patterns.
• NEAT1 acts as a key regulator of porcine skeletal muscle satellite cells, promoting proliferation, and inhibiting differentiation, with distinct subcellular localization patterns.