Identification and functional analysis of Dmrt1 gene and the SoxE gene in the sexual development of sea cucumber, Apostichopus japonicus.

Members of the Doublesex and Mab-3-related transcription factor (Dmrt) gene family handle various vital functions in several biological processes, including sex determination/differentiation and gonad development. Dmrt1 and Sox9 (SoxE in invertebrates) exhibit a very conserved interaction function during testis formation in vertebrates. However, the dynamic expression pattern and functional roles of the Dmrt gene family and SoxE have not yet been identified in any echinoderm species. Herein, five members of the Dmrt gene family (Dmrt1, 2, 3a, 3b and 5) and the ancestor SoxE gene were identified from the genome of Apostichopus japonicus. Expression studies of Dmrt family genes and SoxE in different tissues of adult males and females revealed different expression patterns of each gene. Transcription of Dmrt2, Dmrt3a and Dmrt3b was higher expressed in the tube feet and coelomocytes instead of in gonadal tissues. The expression of Dmrt1 was found to be sustained throughout spermatogenesis. Knocking-down of Dmrt1 by means of RNA interference (RNAi) led to the downregulation of SoxE and upregulation of the ovarian regulator foxl2 in the testes. This indicates that Dmrt1 may be a positive regulator of SoxE and may play a role in the development of the testes in the sea cucumber. The expression level of SoxE was higher in the ovaries than in the testes, and knocking down of SoxE by RNAi reduced SoxE and Dmrt1 expression but conversely increased the expression of foxl2 in the testes. In summary, this study indicates that Dmrt1 and SoxE are indispensable for testicular differentiation, and SoxE might play a functional role during ovary differentiation in the sea cucumber.

Sexually dimorphic expression of foxl2 in the sea urchin (Mesocentrotus nudus).

Sea urchin (Mesocentrotus nudus) is an important economically mariculture species in several Asian countries, and gonads are the sole edible parts for people. In addition to commercial value, it is an excellent model for studying gonadal development, sex determination and sex differentiation. Identify sex-related genes is an effective way to reveal the molecular mechanism of gonadal development. In the present study, the foxl2 homologous gene was identified in M. nudus. Foxl2 is not a maternal factor, and is detected for the first time in two-arm stages. Additionally, the expression of foxl2 in the testis is higher than in the ovaries at the same developmental stages. The foxl2 transcripts were specifically enriched in the cytoplasm of germ cellsboth in the ovary and testis, but their proteins were more concentrated in the area near the oocyte nucleus. Overall, this study contributes to our understanding of the dynamic and sexually dimorphic expression pattern of foxl2 and provide a useful germ cell marker during gametogenesis in sea urchin.

Molecular Cloning and Sexually Dimorphic Expression Analysis of nanos2 in the Sea Urchin, Mesocentrotus nudus.

Sea urchin (Mesocentrotus nudus) is an economically important mariculture species in China and the gonads are the solely edible parts to human. The molecular mechanisms of gonad development have attracted increasing attention in recent years. Although the nanos2 gene has been identified as a germ cell marker in several invertebrates, little is known about nanos2 in adult sea urchins. Hereinto, we report the characterization of Mnnano2, an M. nudus nanos2 homology gene. Mnnanos2 is a maternal factor and can be detected continuously during embryogenesis and early ontogeny. Real-time quantitative PCR (RT-qPCR) and section in situ hybridization (ISH) analysis revealed a dynamic and sexually dimorphic expression pattern of Mnnano2 in the gonads. Its expression reached the maximal level at Stage 2 along with the gonad development in both ovary and testis. In the ovary, Mnnanos2 is specifically expressed in germ cells. In contrast, Mnnanos2 is expressed in both nutritive phagocytes (NP) cells and male germ cells in testis. Moreover, knocking down of Mnnanos2 by means of RNA interference (RNAi) reduced nanos2 and boule expression but conversely increased the expression of foxl2. Therefore, our data suggest that Mnnanos2 may serve as a female germ cell marker during gametogenesis and provide chances to uncover its function in adult sea urchin.

Optimizing a desirable fare structure for a bus-subway corridor.

This paper aims to optimize a desirable fare structure for the public transit service along a bus-subway corridor with the consideration of those factors related to equity in trip, including travel distance and comfort level. The travel distance factor is represented by the distance-based fare strategy, which is an existing differential strategy. The comfort level one is considered in the area-based fare strategy which is a new differential strategy defined in this paper. Both factors are referred to by the combined fare strategy which is composed of distance-based and area-based fare strategies. The flat fare strategy is applied to determine a reference level of social welfare and obtain the general passenger flow along transit lines, which is used to divide areas or zones along the corridor. This problem is formulated as a bi-level program, of which the upper level maximizes the social welfare and the lower level capturing traveler choice behavior is a variable-demand stochastic user equilibrium assignment model. A genetic algorithm is applied to solve the bi-level program while the method of successive averages is adopted to solve the lower-level model. A series of numerical experiments are carried out to illustrate the performance of the models and solution methods. Numerical results indicate that all three differential fare strategies play a better role in enhancing the social welfare than the flat fare strategy and that the fare structure under the combined fare strategy generates the highest social welfare and the largest resulting passenger demand, which implies that the more equity factors a differential fare strategy involves the more desirable fare structure the strategy has.