Abstract

EsDmrt-like is a novel testis-specific Dmrt gene identified from the Chinese mitten crab Eriocheir sinensis. To explore its biological functions, we performed RNA interference (RNAi) by injection of long double-stranded RNA (dsRNA) targeting the coding region of the EsDmrt-like gene. At 24h post-injection with EsDmrt-like-dsRNA, the relative expression level of EsDmrt-like mRNA significantly decreased by 80% (P<0.01) compared to negative control of PBS group, while no RNAi effect was detected using a dsRNA of the green fluorescent protein (GFP), indicating target sequence specificity of this interference effect. Although the EsDMRT-like protein could be obviously detected in the PBS group, its expression in the EsDmrt-like-dsRNA group disappeared in four of the five tested individuals. Taken together, the expressions of EsDmrt-like mRNA and its protein were successfully inhibited by RNAi. After extending RNAi treatment for one month, the crab testicular development exhibited inhibitory effects. The average testis size was reduced and the gonadosomatic index (GSI) was significantly decreased by 75% compared with the controls. The histological and immunohistochemical analysis showed that the spermatogenesis was blocked, with rare spermatozoa in the seminiferous tubules, and the immunohistochemical signal in the spermatogonia and spermatocytes became extremely weak in the EsDmrt-like-dsRNA group. These data demonstrated a critical role for EsDmrt-like in testicular development and spermatogenesis in E. sinensis, and in vivo repetitive injection of EsDmrt-like-dsRNA could work efficiently to inhibit male testicular development. Statement of relevanceThe Chinese mitten crab Eriocheir sinensis is one of the most important aquaculture species and has high commercial value as a food source. EsDmrt-like is a novel testis-specific Dmrt gene identified from the E. sinensis. We performed RNAi by repetitive injection of long double-stranded RNA (dsRNA) targeting the coding region of the EsDmrt-like gene, and certified that EsDmrt-like plays a critical role during spermatogenesis and testicular development.

Full Text
Paper version not known

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call

Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.