Doublesex gene expression of Daphnia pulex during different modes of reproduction by using whole mount in situ hybridization and quantitative expression

Abstract

To analyze the expression pattern of Doublesex( dsx) of Daphnia pulex in different reproductive modes,w e extracted total RNA of different reproductive modes ' Daphnia pulex. Using RT-PCR and Quantitative real-time PCR analyzed dsx expression in different reproductive modes' daphnias. In addition,w e synthesized DIG labeled RNA probe of dsx by in vitro transcription and used w hole mount in situ hybridization researching dsx location expression in different reproductive modes' daphnia. Results show ed that dsx in male and female had no gender specificity of alternative splicing,only a same transcription. Realtime PCR show ed dsx highest expression level w as in male,w hich w as parthenogenetic female's 2. 6 times and 4. 9 times that of the female. They had significant difference( P 0. 05) and gender differences in expression each other. Whole mount in situ hybridization found that dsx gene w as expressed in the somatic tissure in different from reproductive modes ' daphnia,but the difference in quantity and parts. The expression also show ed the gender differences in male and female. In male the signals w ere revealed in the first antenna,the first thoracic limb and the compound eye. The parthenogenetic female and sexual female corresponding sites' expressions w ere relatively w eak. These suggested that Doublesex gene likely played a significant role in alternation of generation and sexual differentiation.