title: Heterozygous mutation of eEF1A1b resulted in spermatogenesis arrest and infertility in male tilapia, Oreochromis niloticus.
abstract: Eukaryotic elongation factor 1 alpha (eEF1A) is an essential component of the translational apparatus. In the present study, eEF1A1b was isolated from the Nile tilapia. Real-time PCR and Western blot revealed that eEF1A1b was expressed highly in the testis from 90 dah (days after hatching) onwards. In situ hybridization and immunohistochemistry analyses showed that eEF1A1b was highly expressed in the spermatogonia of the testis. CRISPR/Cas9 mediated mutation of eEF1A1b resulted in spermatogenesis arrest and infertility in the F0XY fish. Consistently, heterozygous mutation of eEF1A1b (eEF1A1b+/-) resulted in an absence of spermatocytes at 90 dah, very few spermatocytes, spermatids and spermatozoa at 180 dah, and decreased Cyp11b2 and serum 11-ketotestosterone level at both stages. Further examination of the fertilization capacity of the sperm indicated that the eEF1A1b+/-XY fish were infertile due to abnormal spermiogenesis. Transcriptomic analyses of the eEF1A1b+/-testis from 180 dah XY fish revealed that key elements involved in spermatogenesis, steroidogenesis and sperm motility were significantly down-regulated compared with the control XY. Transgenic overexpression of eEF1A1b rescued the spermatogenesis arrest phenotype of the eEF1A1b+/-testis. Taken together, our data suggested that eEF1A1b is crucial for spermatogenesis and male fertility in the Nile tilapia.
authors.affiliation: Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing, 400715, China.
authors.forename: Jinlin
authors.initials: J
authors.lastname: Chen
authors.affiliation: Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing, 400715, China.
authors.forename: Dongneng
authors.initials: D
authors.lastname: Jiang
authors.affiliation: Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing, 400715, China.
authors.forename: Dejie
authors.initials: D
authors.lastname: Tan
authors.affiliation: Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing, 400715, China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Fan
authors.affiliation: Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing, 400715, China.
authors.forename: Yingying
authors.initials: Y
authors.lastname: Wei
authors.affiliation: Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing, 400715, China.
authors.forename: Minghui
authors.initials: M
authors.lastname: Li
authors.affiliation: Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Sciences, Southwest University, Chongqing, 400715, China.
authors.forename: Deshou
authors.initials: D
authors.lastname: Wang
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D023061:Gene Order; D018390:Gene Targeting; D056726:Genetic Association Studies; D056426:Genetic Loci; D006066:Gonads / Q000378:metabolism; D006579:Heterozygote; D007248:Infertility, Male / Q000235:genetics* / Q000378:metabolism; D008297:Male; D009154:Mutation; D020648:Peptide Elongation Factor 1 / Q000235:genetics* / Q000378:metabolism; D021381:Protein Transport; D013091:Spermatogenesis / Q000235:genetics*; D017210:Tilapia / Q000235:genetics* / Q000378:metabolism; D059467:Transcriptome
chemical_list: D020648:Peptide Elongation Factor 1