{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T10:33:03Z","timestamp":1788345183827,"version":"build-2803163510"},"reference-count":32,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,12,15]],"date-time":"2022-12-15T00:00:00Z","timestamp":1671062400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Endocrinol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Growing evidence has indicated that epigenetic factors might be associated with the pathophysiology of idiopathic nonobstructive azoospermia (iNOA). As the most common RNA modification, N6-methyladenosine (m<jats:sup>6<\/jats:sup>A) methylation has recently attracted more attention in the regulation of spermatogenesis; however, its role in the mechanisms of iNOA is still unclear.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objective<\/jats:title><jats:p>To determine the differential expression of mRNA and m<jats:sup>6<\/jats:sup>A methylation status in the testes of iNOA patients.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Testes tissues from diagnosed iNOA and controlled obstructive azoospermia (OA) patients were collected and grouped according to the histological examinations. Total RNA was isolated and quantified by an m<jats:sup>6<\/jats:sup>A RNA Methylation Quantification Kit. The expression level of mRNAs was detected by qRT-PCR analysis. Differentially expressed m<jats:sup>6<\/jats:sup>A genes were analyzed using the human ArrayStar m<jats:sup>6<\/jats:sup>A epitranscriptomic microarray, and bioinformatics analyses were applied.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>A total of 36 iNOA and 8 controlled patients were included. The global expression of m<jats:sup>6<\/jats:sup>A in the iNOA group was significantly decreased. A dosage relationship was observed between the m<jats:sup>6<\/jats:sup>A decline and the degree of impaired spermatogenesis, with the successive process of normal spermatogeneis, hypospermatogenesis (HP), maturation arrest (MA), and Sertoli cell-only syndrome (SO). Four down-expressed genes (<jats:italic>BDNF, TMEM38B, RPL3L, and C22orf42<\/jats:italic>) displayed significantly lower expression of m<jats:sup>6<\/jats:sup>A methylation. Additionally, they also showed a gradually down-expressed tendency in the three groups (OA, HP, SO\/MA groups). Moreover, m<jats:sup>6<\/jats:sup>A reader EIF3A was approved to have differential expression through microarrays analysis, which was consistent with the result from the qRT-PCR test.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>The m<jats:sup>6<\/jats:sup>A expression was gradually downregulated in the testes tissue from iNOA patients in accordance with the degree of spermatogenic dysfunction. The determined differential expression of mRNA and m<jats:sup>6<\/jats:sup>A methylation status may represent potentially novel molecular targets for the mechanism study of iNOA in the epigenetic level, which could benefit the understanding of the pathophysiology of iNOA.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fendo.2022.1063929","type":"journal-article","created":{"date-parts":[[2022,12,15]],"date-time":"2022-12-15T08:02:29Z","timestamp":1671091349000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":6,"title":["Joint analysis of m6A and mRNA expression profiles in the testes of idiopathic nonobstructive azoospermia 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