Title The expression characteristics of FAM71D and its association with sperm motility
Authors Ma, Qian
Li, Yuchi
Luo, Manling
Guo, Huan
Lin, Shouren
Chen, Jianbo
Du, Ye
Jiang, Zhimao
Gui, Yaoting
Affiliation Peking Univ, Shenzhen Hosp, Guangdong Key Lab Male Reprod Med & Genet, Inst Urol,Shenzhen PKU HKUST Med Ctr, Shenzhen 518036, Peoples R China.
Keywords FAM71D
sperm flagella
calmodulin
sperm motility
asthenozoospermia
MALE-INFERTILITY
MAMMALIAN SPERM
MICE DEFICIENT
MOUSE SPERM
SPERMATOZOA
CALMODULIN
CELLS
FLAGELLUM
RNA
FERTILIZATION
Issue Date 2017
Publisher HUMAN REPRODUCTION
Citation HUMAN REPRODUCTION. 2017, 32(11), 2178-2187.
Abstract STUDY QUESTION: What are the features of FAM71D (Family with sequence similarity 71, member D) expression and is there an association between FAM71D expression and sperm motility? SUMMARY ANSWER: FAM71D, a novel protein exclusively expressed in the testis, is located in sperm flagella and is functionally involved in sperm motility. WHAT IS KNOWN ALREADY: Some testis-specific proteins have been reported as potential diagnostic biomarkers to evaluate the spermatogenesis process and sperm quality. We have identified a novel testis-specific protein, FAM71D, through microarray data analysis, yet little is known about its expression and function. STUDY DESIGN, SIZE, DURATION: FAM71D mRNA and protein expression was quantified during mouse testis development. Its localization in germ cells was detected by dual-labeled immunostaining in testis sections and sperm smears. The clinical significance was assessed by comparing FAM71D expression in spermatozoa from normozoospermic controls and asthenozoospermic patients. PARTICIPANTS/MATERIALS, SETTING, METHODS: Testes were dissected from C57BL/6 J male mice at postnatal ages of 1, 2, 3, 4, 6, 8 weeks and 6 months, and sperm was collected from cauda epididymides of adult mice by the swim-up method. Human spermatozoa were isolated from 100 human semen samples by density gradient Percoll centrifugation. RT-qPCR and western blot were performed to semi-quantify the expression of FAM71D in mouse testis, and in the ejaculated spermatozoa of normozoospermic controls and asthenozoospermic patients. Immunofluorescence staining was used to detect the localization of FAM71D. Co-immunoprecipitation assay was performed to evaluate the interaction between FAM71D and calmodulin. An antibody blocking assay was employed to assess the role of FAM71D in sperm motility. MAIN RESULTS AND THE ROLE OF CHANCE: Our results showed that FAM71D was exclusively expressed in the testis in an age-dependent manner. FAM71D expression exhibited dynamic change in the cytoplasm of spermatids during spermiogenesis and was finally retained in sperm flagella. FAM71D could interact with calmodulin. Use of anti-FAM71D antibody on sperm significantly decreased sperm motility. Expression level of FAM71D was markedly reduced in the ejaculated spermataozoa of asthenozoospermic patients (P < 0.05), and this was correlated with sperm progressive motility (r = 0.7435, P < 0.0001). LARGE SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: The sample size was limited and it is necessary to verify the correlation of FAM71D expression with sperm motility in larger cohorts. Furthermore, our results were descriptive and follow-up studies would be needed to elucidate the detailed role of FAM71D in sperm motility. WIDER IMPLICATIONS OF THE FINDINGS: This is the first systematic study to document the expression of endogenous FAM71D and a function for FAM71D in sperm motility. It provides new insights into our understanding of sperm motility regulation and causes of male infertility. STUDY FUNDING/COMPETING INTERESTS: This study was funded by the National Natural Science Foundation of China, Guangdong Natural Science Foundation and the Shenzhen Project of Science and Technology. The authors have no competing interests.
URI http://hdl.handle.net/20.500.11897/484545
ISSN 0268-1161
DOI 10.1093/humrep/dex290
Indexed SCI(E)
Appears in Collections: 深圳医院

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