Lymnaea stagnalis neuronal transcriptome
Lymnaea stagnalis neuronal transcriptome
We developed a reduced representation sequencing method that can detect low abundant transcripts. Our method greatly reduces the amount of sequencing with more accurate quantification. Basically, sequential addition of adaptors paired with controlled digestion of cDNA generates a one transcript on read type of analysis. Combined 5’ with 3’ targeted libraries produces greater coverage of individual transcripts and high quality assembly.
The freshwater pond snail, Lymnaea stagnalis is an important model to study mechanisms of synapse formation and cellular bases of behavior. However, there is very limited information about the molecular organization of its nervous system. Here, using pyrosequencing approach we performed analysis of transcripts expressed in the central nervous system of Lymnaea. Specifically, we developed a reduced representation sequencing method that can detect low abundant transcripts. Our method greatly reduces the amount of sequencing with more accurate quantification. Basically, sequential addition of adaptors paired with controlled digestion of cDNA generates a one transcript on read type of analysis. Combined 5’ with 3’ targeted libraries produces greater coverage of individual transcripts and high quality assembly.   Our initial analysis allowed us to identify more than 20,000 of putative neuronal transcripts including a diversity of novel neuropeptides, receptors, ionic channels, and other components of signal transduction pathways essential for future analysis of molecular bases of neuronal functions in this model organism.