Establishment of three estrogen receptors (esr1, esr2a, esr2b) knockout lines for functional study in Nile tilapia. Issue 191 (July 2019)

• Record Type: Journal Article
• Authors: Yan, Longxia; Feng, Haiwei; Wang, Feilong; Lu, Baoyue; Liu, Xingyong; Sun, Lina; Wang, Deshou
• Is Part Of: Journal of steroid biochemistry and molecular biology. Issue 191(2019)
• Publication Date: 2019-07
• Subjects: dah days after hatching -- E2 17β-estradiol -- 11-KT 11-ketotestosterone -- IHC immunohistochemistry -- CRISPR/Cas9 custered regularly interspaced short palindromic repeats/CRISPR associated -- gRNA guide RNA -- GSI gonadosomatic index -- RT-PCR reverse transcription PCR -- EIA enzyme immunoassay -- VCL curvilinear velocity -- VSL straight linear velocity -- BCF flagellum beat frequency; Three estrogen receptors -- Mutation -- CRISPR/Cas9 -- Phenotype -- Differential functions -- Nile tilapia; Steroid hormones -- Periodicals; Biochemistry -- Periodicals; Hormones -- Periodicals; Molecular Biology -- Periodicals; Hormones stéroïdes -- Périodiques; Steroid hormones; Periodicals; 572.579
• DOI: 10.1016/j.jsbmb.2019.105379
• Languages: English
• ISSNs: 0960-0760

Abstract

Highlights: We established three esr mutant lines and analyzed their phenotypes in tilapia. Mutation of esr1 displayed normal oogenesis and spermatogenesis at different developing stages. Mutation of esr2a displayed retarded follicle growth in female and reduced fertility in male. Mutation of esr2b was completely infertile due to reproductive tracts malformation in both female and male. Abstract: Estrogens play fundamental roles in regulating reproductive activities and they act through estrogen receptors ( ESR s) in all vertebrates. To date, distinct roles of estrogen receptors have been characterized only in human and model organisms, including mouse, rat, zebrafish and medaka. Physiological role of estrogen/receptor signaling in reproduction remains poorly defined in non-model organisms. In the present study, we successfully generated esr1, esr2a and esr2b mutant lines in tilapia by CRISPR/Cas9 and examined their phenotypes. Surprisingly, the esr1 mutants showed no phenotypes of reproductive development and function in both females and males. The esr2a mutant females showed significantly delayed ovarian development and follicle growth at 90 and 180 dah, and the development caught up later at 360 dah. The esr2a mutant males showed no phenotypes at 90 dah, and displayed smaller gonads and efferent ducts, less spermatogonia and more abnormal sperms at 180 dah. In contrast, the esr2b mutants displayed abnormal development of ovarian ducts and efferent ducts which failed to connect to the genital orifice, and which in turn, resulted in infertility in female and male, respectively, although they produced gametes in their gonads. Taken together, our study provides evidence for differential functions of esr1, esr2a and esr2b in fish reproduction.