LAUSR

Lactate produced by glycolysis in Sertoli cells (SCs) is the main energy substrate for developing germ cells and plays a vital role in spermatogenesis. MicroRNAs (miRNAs) function as posttranscriptional regulators of gene expression in biological processes. We have previously shown that hyperthermia (43°C, 30 min) promotes lactate secretion by inhibiting phosphorylation of adenosine monophosphate‐activated protein kinase (AMPK) in cultured immature boar SCs. However, it is unclear whether miRNAs are involved in AMPK‐modulated glycolysis in SCs. In the present study, we identified 349 miRNAs (227 upregulated and 122 downregulated) in hyperthermia‐treated boar SCs by next‐generation high‐throughput RNA sequencing. MiR‐8‐3p, which was found to be a novel upregulated miRNA in hyperthermia‐treated SCs, suppressed the expression of AMPK upstream genes (protein phosphatase 2 subunit B, PPP2R5B), and further downregulated the expression of p‐AMPK. The miR‐8‐3p mimic upregulated expression of glucose transporter 3, lactate dehydrogenase A and monocarboxylate transporter 1, and increased lactic acid dehydrogenase activity, lactate secretion, and ATP depletion in SCs; the miR‐8‐3p inhibitor had the opposite effects on these parameters. Our findings indicate that miR‐8‐3p acts as a novel regulator of AMPK‐modulated lactate secretion by targeting PPP2R5B in hyperthermic boar SCs.