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The present study aimed to investigate serine protease 55 (PRSS55), which was reported to play a role in sperm\u2010uterotubal junction (UTJ) migration and sperm\u2010zona pellucida (ZP) binding. We found that PRSS55 was specifically expressed in testicular spermatids and epididymal spermatozoa. By constructing knockout mice targeting all transcripts of <jats:italic>Prss55<\/jats:italic>, we demonstrated that deletion of <jats:italic>Prss55<\/jats:italic> resulted in a serious decline of male fertility, with significantly increased sperm malformation and decreased sperm motility. In <jats:italic>Prss55<\/jats:italic><jats:sup>\u2212\/\u2212<\/jats:sup> mice, increased structural abnormality, including deficient \u201c9\u00a0+\u00a02\u201d microtubules, damaged peripheral dense fibre, and defective mitochondrial cristae, were found in sperm. In addition, sperm showed decreased expression of electron transfer chain molecules and lower ATP contents. These could be the potential causes of the astheno\/teratozoospermia phenotype of the <jats:italic>Prss55<\/jats:italic><jats:sup>\u2212\/\u2212<\/jats:sup> mice, and provided new evidence for the previously reported impaired sperm\u2010UTJ migration. Moreover, preliminary studies allowed us to speculate that PRSS55 might function by activating type II muscle myosin in the testis, which is involved in many processes requiring motivation and cytoskeleton translocation. Thus, PRSS55 is essential for the structural differentiation and energy metabolism of sperm, and might be a potential pathogenic factor in astheno\/teratozoospermia. Our results provide an additional explanation for the male sterility of <jats:italic>Prss55<\/jats:italic><jats:sup>\u2212\/\u2212<\/jats:sup> mice, and further reveal the role of PRSS55.<\/jats:p>","DOI":"10.1111\/jcmm.16116","type":"journal-article","created":{"date-parts":[[2021,1,13]],"date-time":"2021-01-13T19:07:16Z","timestamp":1610564836000},"page":"2040-2051","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":28,"title":["PRSS55 plays an important role in the structural differentiation and energy metabolism of sperm and is required for male fertility in mice"],"prefix":"10.1111","volume":"25","author":[{"given":"Feng","family":"Zhu","sequence":"first","affiliation":[{"name":"State Key Laboratory of Reproductive Medicine Department of Histology and Embryology Nanjing Medical University  Nanjing China"},{"name":"Department of Pathology The First People's Hospital of Changzhou  Changzhou 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Embryology Nanjing Medical University  Nanjing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yiqiang","family":"Cui","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Reproductive Medicine Department of Histology and Embryology Nanjing Medical University  Nanjing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaofei","family":"Liu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Reproductive Medicine Department of Histology and Embryology Nanjing Medical University  Nanjing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuejiang","family":"Guo","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Reproductive Medicine Department of Histology and Embryology Nanjing Medical University  Nanjing 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