Single-cell gene expression analysis in C. elegans [electronic resource]
- Sarah Joann Aerni.
- Physical description
- 1 online resource.
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- Aerni, Sarah Joann.
- Batzoglou, Serafim primary advisor.
- Kim, Stuart, primary advisor.
- Paik, David, advisor.
- Stanford University Program in Biomedical Informatics.
- Complex organisms contain many individual cells that express unique sets of genes. Unraveling the full complexity of organisms requires an understanding of the unique gene expression signatures of individual cells. Obtaining these cellular signatures enables a deeper understanding of the underlying biological processes, including development and aging. Model organisms, such as C. elegans, are ideal platforms for single-cell studies, offering true single-cell resolution. In the nematode, C. elegans, each of its 959 somatic cells are uniquely identifiable in every individual. Through high-resolution 3D imaging of in-situ fluorescent reporters, expression levels can be observed in all 959 cells. The key to making this powerful system a viable source of discovery through large-scale, high-resolution gene expression analysis requires automation of the annotation process. This dissertation describes computational approaches to identifying single-cells to obtain high-resolution gene expression levels in adult C. elegans. These approaches include an image processing pipeline enabling both manual and automatic annotation of images. For the latter, I describe a set of automatic annotation methods developed for single-cell identification in 3D confocal images of adult C. elegans utilizing machine learning approaches. These approaches automatically assign a set of labels to cells based on (1) cell-lineage, (2) tissue type or (3) the anatomical region occupied within the worm. Finally, this dissertation describes the types of biological insights researchers can gain from such high-resolution data. In particular, it develops a novel gene expression analysis method that combines the known cell lineage and expression levels of 96 genes in 363 single-cells in the first larval stage of C. elegans. Together, these methods provide C. elegans researchers with a powerful toolkit for performing single-cell gene expression analysis of worms. This work enables studies in the field of development and, for the first time, aging at the single-cell resolution using automated annotation approaches in the adult worm.
- Publication date
- Submitted to the Program in Biomedical Informatics.
- Thesis (Ph.D.)--Stanford University, 2012.
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