The Application Of Neuron-specific Expression System In C. Elegans

Because of simple anatomy, clear genetic background, completion of sequencing,short life cycle and many other advantages, Caenorhabditis elegans has undoubtedly beenused as a model organism to study behavior and neurobiology, Genetics andDevelopmental Biology. Compare with other model organism, neural system of C. elegansis simple. There are only302neurons in neurons system and they have been classified intoseveral groups---motorneuron, sensory neuron and interneuron. Though less those neurons,they generate such complicated connections carrying out significantly indispensiblefounctions according to the developmental orders. Some of neurons and their subtleconnections have been studied by biotechnology, such as biological electron microscopyThe main job of my project is labling C. elegans neurons, expressing functional gene,studying neuronal function. In my project, we combined the gateway technology,site-specific recombinase system (FRT/FLP), inducible expression systems with confocalmicroscopy imaging techniques to label C. elegans neurons. We chose promoters withintersections, so that we can lable neurons by different colors when different promotersexpress different fluorescent proteins. Besides we also generated a combination systemwhich enrolled intersection of promoters and site-specific recombinase system (FRT/FLP)to lable single neurons. We can label the neuron ASH by intersection of promoters Psra-6and Pgpa-11. After target neuron was labeld, some proteins such as ChR2, mito-miniSOG,CatCh, TeTx and CED-3can express in single neurons, so that further studies in neuronfunctions of nervous system of C. elegans can continue.Lots of transgenic assay in vivo or in vitro need regulate the expression, but somereusable expression has its fatal flaws. Tetracycline-inducible expression system isconsidered as the most ideal gene expression system in eukaryotic gene regulatoryexpression system, but it almost had no applications in C. elegans. Usingtetracycline-inducible expression system in our project, we constructed reaction elementsTRE::TagRFP and regulatory elementsPsrh-11::rtTA3::s12TagRFP. Experimental datarevealed that tetracycline-inducible expression system function in controlling RFPexpression in ASJ neurons temporally. Thus, tetracycline-inducible expression system expression system can be utilized in study gene function C. elegans.