Loligo opalescens neuronal transcriptome

The squid, Loligo is a classical model to study mechanisms of exitability. However, little is known about the molecular organization of its nervous system. Here, using pyrosequencing approach we performed analysis of transcripts expressed in the central nervous system of Loligo. 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 major components of signal transduction pathways essential for future analysis of molecular bases of neuronal functions in this model organism.

枪乌贼属（Loligo）是研究兴奋性机制的经典模式生物。然而，目前对其神经系统的分子组织机制尚不清楚。本研究采用焦磷酸测序（pyrosequencing）技术，对枪乌贼中枢神经系统中表达的转录本进行了分析。具体而言，本研究开发了一种可检测低丰度转录本的简化表征测序方法。该方法可大幅降低测序通量，同时实现更精准的定量分析。简言之，通过依次添加接头并辅以可控的互补DNA（cDNA）酶切，本方法可实现单转录本读长型分析。结合5'端与3'端靶向文库，可实现单个转录本的更高覆盖度以及高质量的序列组装。本研究的初步分析已鉴定出该模式生物神经元功能分子基础研究所需的信号转导通路核心组分，为后续相关研究奠定了基础。