Constructing novel, genetically encodable tools to study the role of actin in developmental myelination
Abstract/Contents
- Abstract
- The insulating myelin sheath formed by oligodendrocytes is crucial for rapid conduction of action potentials. The understanding of how oligodendrocytes wrap this specialized membrane is limited. Actin, a key regulator of cell structure and movement, is hypothesized to play an important role (Bauer et al., 2009). Further investigation of the role of actin in myelination necessitates a cell-specific tool with fine temporal and dose- dependent control. We developed genetically encodable actin-modifying tools (gACTs). Using DNA sequences from endogenous actin-modifying proteins or bacterial toxins allowed us to harness their actin modifying power. At this stage, the constructs allow for temporal control and either dose-dependent or cell-specific control, but ultimately the tool will have all three features. To validate the constructs, we transfected HeLa cells and visualized actin morphology. We found clear actin morphology changes upon transfection for all five constructs with variable effectiveness and cytotoxicity. To provide temporal and dose-dependent control, we fused the destabilization domain (DD) to the constructs (Banaszynski et al., 2006). To provide temporal control and cell specificity, we replaced the strong cytomegalovirus (CMV) promoter with a myelin basic protein (MBP) promoter, conferring expression only in mature stages of oligodendrocytes. In addition to elucidating the role of actin in myelination, gACTs are a useful general cell biology tool to study actin in multicellular processes such as development or diseases such as multiple sclerosis.
Description
| Type of resource | text |
|---|---|
| Date created | June 15, 2014 |
Creators/Contributors
| Author | Turan, Julia | |
|---|---|---|
| Advisor | Zuchero, J. Bradley | |
| Advisor | McConnell, Susan K. | |
| Primary advisor | Barres, Ben | |
| Degree granting institution | Stanford University, Department of Biology, 2014 |
Subjects
| Subject | Biology |
|---|---|
| Subject | Neurobiology |
| Subject | Cell Biology |
| Genre | Thesis |
Bibliographic information
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- User agrees that, where applicable, content will not be used to identify or to otherwise infringe the privacy or confidentiality rights of individuals. Content distributed via the Stanford Digital Repository may be subject to additional license and use restrictions applied by the depositor.
- License
- This work is licensed under a Creative Commons Attribution Share Alike 3.0 Unported license (CC BY-SA).
Preferred citation
- Preferred Citation
- Turan, Julia and Zuchero, J. Bradley and Barres, Ben. (2014). Constructing novel, genetically encodable tools to study the role of actin in developmental myelination. Stanford Digital Repository. Available at: http://purl.stanford.edu/zj542zv7504
Collection
Undergraduate Theses, Department of Biology, 2013-2014
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- Contact
- jturan@stanford.edu
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