Project description:In order to identify unexpected, or indeed previously uncharacterized genes may be important in sex- or gonad development, we developed a custom cDNA microarray represent 3837 unique transcripts of Scylla paramamosain derived from our EST project. Thirty-nine putative transcripts were observed to differentially expressed in testis and ovaries (P<0.05).
Project description:In order to identify unexpected, or indeed previously uncharacterized genes may be important in sex- or gonad development, we developed a custom cDNA microarray represent 3837 unique transcripts of Scylla paramamosain derived from our EST project. Thirty-nine putative transcripts were observed to differentially expressed in testis and ovaries (P<0.05). Two-condition experiment, Ovary vs. Testis. Biological replicates: 3 Ovaries, 3 Testis, 2 dye-swaps.
Project description:Background: The Scylla paramamosain is a very important aquaculture crustacean species in the southeast coastal areas of China including Shantou. For the past few years, mud crab cultured in Niutianyang of Shantou suffered from serious diseases, especially the bacterial diseases (such as Vibrio parahaemolyticus). In eukaryotes, small RNAs can regulate gene expression in post-transcription to act on host-pathogen interaction system. Aims: V.parahaemolyticus isolated from Shantou Niutianyang crab culture area was injected to S.paramamosains to carry out an essential analysis on global miRNA expression in diverse tissues between two groups by the Illumina Solex deep sequencing technology. Methodology:To examine the relationship between mud crab miRNA expression and the bacterial pathogen, we collected mixed two pools of equal amounts of RNA from 7 different mud crab tissues (mesenteron, heart, liver, gill, brain, muscle and blood) and sequencing by Illumine/Solexa deep sequencing technology under normal conditions and during infection with V.parahaemolyticus. The high throughput sequencing resulted in 19,144,358 and 18,559,070 raw reads corresponding to 17,496,577 and 16,888,096 high-quality mappable reads for the normal and infected mixed pools, respectively. Stem-loop RT-qPCRs were used to confirm the microRNAs expression in different tissues of two pools. The results show that miRNAs might play a key role in regulating gene expression during mud crab S.paramamosain infection with V.parahaemolyticus. Conclusions: We identified a large number of miRNAs during the mud crab Scylla paramamosain infection with V.parahaemolyticus, some of which are differentially expressed between the treatments and the controls. The study provides an opportunity for further understanding of small RNA function in the regulation of molecular response and gives us clues for further studies of the mechanisms of V.parahaemolyticus infection in mud crab. Examination of miRNA expression in normal Scylla paramamosain group and the Scylla paramamosain infected with Vibrio parahaemolyticus
Project description:Scylla paramamosain (Crustacea) is a commercially important euryhaline species distributed along the coast of southern China and other Indo-Pacific countries. However, a sudden variation in salinity will cause injury or even death of S. paramamosain. In this paper, we simulated a sudden decrease in salinity due to heavy precipitation in crab ponds. Comparison of gill microstructures of individuals in the control group and decreased salinity group showed gills became shorter and thicker, while the top of the filaments became swollen and then returned to normal after 120 h. A total of 3962 proteins were identified by proteomic sequencing of gills after 120 h under conditions of decreased salinity. 845 proteins were differentially expressed proteins: 371 up-regulated and 474 down-regulated. Of the enriched KEGG pathways, 20 were up-regulated and 14 were down-regulation (p<0.05). Among the significantly enriched up-regulated pathways, six were associated with amino acid metabolism and three were associated with Na+-K+-ATPase enzymatic activities. Pathways associated with redox metabolism and energy metabolism were identified. These results showed that in response to a decrease in salinity, S. paramamosain could adapt to the environment after 120 h. Molecular mechanism of this adaptation involved amino acid metabolism and Na+-K+-ATPase ion transport. Meanwhile, energy metabolism and redox metabolism were critical to the adaptation to a sudden decrease in salinity. This study, for the first time at the protein level, revealed the molecular mechanisms underlying salinity adaption of S. paramamosain and provides theoretical guidelines for the cultivation of S. paramamosain and other marine crustaceans.