Molecular Cloning, Expression, and In Situ Hybridization Analysis of Forkhead Box Protein L2 during Development in Macrobrachium nipponense

Abstract

Forkhead box protein L2 (Foxl2) has dramatic effects on early differentiation and development of the female gonad in mammals and fish through the repression of SOX9 expression. In this study, we aimed to isolate a full‐length cDNA sequence encoding Foxl2 from the Macrobrachium nipponense (Mn‐Foxl2) and investigated its gene function. The full‐length cDNA sequence of Mn‐Foxl2 was 2097 bp with an open reading frame of 1740 bp, encoding 579 amino acids. Phylogenetic analysis revealed considerable evolutionary divergence between Mn‐Foxl2 and Foxl2 from fish. Mn‐Foxl2 may be involved in gonadal sex differentiation, based on its dramatically high expression during the sex‐differentiation sensitive period. Tissue distributions indicated that Mn‐Foxl2 mRNA expression was higher in the testis than that in the ovary and higher in males than in females in the same tissues, implying that Foxl2 may promote both male and female sexual development in M. nipponense. In situ hybridization analysis and quantitative polymerase chain reaction analysis in the gonad during the reproductive cycle further confirmed the important roles of Foxl2 in early gonad development in M. nipponense. This study advances our understanding of the role of Foxl2 in M. nipponense and provides the basis for further studies of Foxl2 in crustaceans.

Investigation of novel functions of three genes in oriental river prawn,Macrobrachium nipponense: Molecular Cloning, Expression, andIn situHybridization Analysis

“…As described in detail previously [13], healthy adult M. nipponense with wet weight of 3.785-26g were obtained from Tai Lake in Wuxi, China (120°13’44“E, 31°28’22”N). These specimens were maintained in aerated freshwater under lab conditions at the temperature of 28°C for at least 72 h prior to tissue collection.…”

“…As described in detail previously [13], total RNA was extracted from androgenic gland as template using RNAiso Plus Reagent (Takara Bio Inc.), followed the protocol of the manufacturer. The RNase-free DNase I (Sangon, Shanghai, China) was used to treat the isolated RNA to eliminate possible genomic DNA contamination.…”

“…As described in detail previously [13], a M-MLV reverse transcriptase was used to perform the first strand 3′cDNA and 5′cDNA synthesis for gene cloning using the 3′-Full RACE Core Set Ver.2.0 kit and the 5′-Full RACE kit (Takara Bio Inc., Japan), respectively with the reaction conditions recommended by the manufacturer. The synthesized cDNAs were kept at -20°C.…”

Progress in Aquaculture Genetics and Breeding in China

“…Other papers in this special issue are focused on the characterization of important genes potentially affecting aquaculture performance in various organisms (Jin et al ; Ping et al ; Shan et al ; H. Zhang, Zhang et al ; H. Zhang, Zhao et al ; X. Zhang et al ). Shan et al () cloned the crustacean cardioactive peptide (CCAP) gene from eyestalks of the oriental river prawn, Macrobrachium nipponense , and studied its expression profile under hypoxic conditions.…”

“…Their results suggested that Mn‐Foxl2 may be involved in morphological larval changes, initiation of metamorphosis, and gonadal sex differentiation. Mn‐Foxl2 mRNA expression levels were significantly higher in testes than in ovaries, implying that Mn‐Foxl2 may promote both the male and female sexual development in M. nipponense (Jin et al ).…”