title: Identification of seven genes essential for male fertility through a genome-wide association study of non-obstructive azoospermia and RNA interference-mediated large-scale functional screening in Drosophila.
abstract: Non-obstructive azoospermia (NOA) is a complex and severe condition whose etiology remains largely unknown. In a genome-wide association study (GWAS) of NOA in Chinese men, few loci reached genome-wide significance, although this might be a result of genetic heterogeneity. Single nucleotide polymorphisms (SNPs) without genome-wide significance may also indicate genes that are essential for fertility, and multiple stage validation can lead to false-negative results. To perform large-scale functional screening of the genes surrounding these SNPs, we used in vivo RNA interference (RNAi) in Drosophila, which has a short maturation cycle and is suitable for high-throughput analysis. The analysis found that 7 (31.8%) of the 22 analyzed orthologous Drosophila genes were essential for male fertility. These genes corresponded to nine loci. Of these genes, leukocyte-antigen-related-like (Lar) is primarily required in germ cells to sustain spermatogenesis, whereas CG12404, doublesex-Mab-related 11E (dmrt11E), CG6769, estrogen-related receptor (ERR) and sulfateless (sfl) function in somatic cells. Interestingly, ERR and sfl are also required for testis morphogenesis. Our study thus demonstrates that SNPs without genome-wide significance in GWAS may also provide clues to disease-related genes and therefore warrant functional analysis.
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing 210029, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Yu
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing 210029, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Wu
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Epidemiology and Biostatistics, School of Public Health, Nanjing Medical University, Nanjing 211166, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Wen
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing 210029, China.
authors.forename: Yujuan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing 210029, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Epidemiology and Biostatistics, School of Public Health, Nanjing Medical University, Nanjing 211166, China.
authors.forename: Bixian
authors.initials: B
authors.lastname: Ni
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Epidemiology and Biostatistics, School of Public Health, Nanjing Medical University, Nanjing 211166, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Lin
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Epidemiology and Biostatistics, School of Public Health, Nanjing Medical University, Nanjing 211166, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Dong
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing 210029, China.
authors.forename: Zuomin
authors.initials: Z
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Epidemiology and Biostatistics, School of Public Health, Nanjing Medical University, Nanjing 211166, China ctong@zju.edu.cn shajh@njmu.edu.cn guo_xuejiang@njmu.edu.cn zhibin_hu@njmu.edu.cn.
authors.forename: Zhibin
authors.initials: Z
authors.lastname: Hu
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing 210029, China, ctong@zju.edu.cn shajh@njmu.edu.cn guo_xuejiang@njmu.edu.cn zhibin_hu@njmu.edu.cn.
authors.forename: Xuejiang
authors.initials: X
authors.lastname: Guo
authors.affiliation: State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing 210029, China, ctong@zju.edu.cn shajh@njmu.edu.cn guo_xuejiang@njmu.edu.cn zhibin_hu@njmu.edu.cn.
authors.forename: Jiahao
authors.initials: J
authors.lastname: Sha
authors.affiliation: Life Sciences Institute and Innovation Center for Cell Biology, Zhejiang University, Hangzhou 310058, China ctong@zju.edu.cn shajh@njmu.edu.cn guo_xuejiang@njmu.edu.cn zhibin_hu@njmu.edu.cn.
authors.forename: Chao
authors.initials: C
authors.lastname: Tong
mesh_terms: D000818:Animals; D044466:Asian People / Q000235:genetics; D053713:Azoospermia / Q000235:genetics*; D004195:Disease Models, Animal; D004330:Drosophila / Q000235:genetics*; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D005298:Fertility / Q000235:genetics*; D056726:Genetic Association Studies; D056426:Genetic Loci; D020022:Genetic Predisposition to Disease; D006801:Humans; D008297:Male; D020641:Polymorphism, Single Nucleotide; D034622:RNA Interference; D011960:Receptors, Estrogen / Q000235:genetics / Q000378:metabolism; D013091:Spermatogenesis / Q000235:genetics; D015238:Sulfotransferases / Q000235:genetics / Q000378:metabolism; D013737:Testis / Q000378:metabolism
chemical_list: D029721:Drosophila Proteins; C475538:ERR protein, Drosophila; D011960:Receptors, Estrogen; D015238:Sulfotransferases; C500949:sfl protein, Drosophila