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 F0 XY 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.
Highlights
Eukaryotic elongation factor 1 alpha is an essential component of the translational apparatus
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
Only one Eukaryotic elongation factor 1 alpha (eEF1A) gene has been reported in zebrafish (Danio rerio), sea bream (Sparus aurata), Nile tilapia (Oreochromis niloticus) and medaka (Oryzias latipes)[5,6,7,8]
Summary
Eukaryotic elongation factor 1 alpha (eEF1A) is an essential component of the translational apparatus. Eukaryotic translation elongation factor 1A (eEF1A) is one of the most abundant protein synthesis factors in eukaryotic cells. Mutation of the elongation factors for proteins synthesis, may result in infertility in vertebrates, as was demonstrated for eEF420. Besides its canonical function in translation elongation, eEF1A is involved in other cellular processes, such as severing microtubule[21], bundling F actin[22], controlling cell apoptosis[23] and regulating protein degradation[24,25]. EEF1A might be directly involved in gametogenesis via protein synthesis in germ cells. Species with different eEF1A genes expressed in the testis and ovary would be a good model to study the diverse function of the eEF1A genes in spermatogenesis and oogenesis
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