Improved single cell RT-PCR for investigatingmolecular identity of muscle nociceptor

• Main Authors: Hung-Chin Wang, 王宏晉
• Other Authors: Chih-Cheng Chen
• Format: Others
• Language: en_US
• Published: 2007
• Online Access: http://ndltd.ncl.edu.tw/handle/07106259830129792046

碩士 === 國立臺灣大學 === 動物學研究研究所 === 95 === Single cell RT-PCR is a powerful tool to explore the molecular identity of single cells. It can determine the expression of many genes in a single cell simultaneously and help to characterize the diverse cell population like dorsal root ganglion DRG neurons. However, there are still some limitations of single cell RT-PCR to obtain a large amount of informative data in gene expression from a single cell. Therefore, I established a new protocol to perform single cell RT-PCR with modification on cell harvesting approach, increased sensitivity of gene determination and reduced contamination. Ideally, this method will be able to detect over 100 genes in a single neuron. I applied this modified single cell RT-PCR to investigate the molecular identity of muscle afferent neurons and showed that ASIC3 expressing muscle afferent neurons had distinct molecular identity from other neurons. The single cell RT-PCR data were consistent with previous studies using immunostaining. In the thesis, I successfully established a powerful technique to investigate the molecular identity of a diverse neuronal population.