What do microglia do? New tools for the study of rodent and human microglia
Abstract/Contents
- Abstract
- The specific function of microglia, the tissue resident macrophages of the brain and spinal cord, has been difficult to ascertain due to a lack of tools that distinguish microglia from other immune cells, thereby limiting their specific immunostaining, purification, and manipulation. Due to their unique developmental origins and predicted functions, the distinction of microglia from other myeloid cells is critically important to understanding brain development and disease; better tools would greatly facilitate studies of microglia function in the developing, adult, and injured CNS. This thesis consists of two main parts. The first is dedicated to identifying Transmembrane Protein 119 (Tmem119), a cell surface protein of unknown function, as a highly expressed microglia-specific marker in both mouse and human. In this section, I provide background information about microglia, the current state of research, and the importance of a molecular marker of microglia to better understand their function. I then discuss the qualities important for a unique molecular marker and show that, by RNA, Tmem119 fulfills these in both mouse and human. I discuss the development of monoclonal antibodies to the intracellular and extracellular domains of Tmem119 that together enable the immunostaining of microglia in histological sections in healthy and diseased brains and the isolation of mouse microglia by FACS. In the second part, I use these antibodies to generate the first RNAseq profiles of highly pure mouse microglia during development and after an immune challenge. I optimized an isolation protocol that reduces baseline microglial activation to better capture the in vivo genetic profiles of microglia. I found that mouse microglia mature by the second postnatal week, and that this maturation both corresponds with Tmem119 immunoreactivity and correlates with the expression of many microglia-enriched genes. I identified genes which are differentially expressed during development, after immune challenge, or between microglia and other macrophages. I generated several lists of genes containing enriched transcription factors, receptor-ligands, and disease-related genes. I also generated a resource, available at www.BrainRNAseq.org for scientists to mine these data themselves. Through these analyses, we made several new predictions about the functions of microglia - including a potential role in hemostasis. Finally, in the closing chapters, I discuss future directions and some preliminary work to harness these insights for future work - understanding the function of this highly specific and enriched gene, developing genetic tools to study microglia in vivo, and ultimately - to better understand what microglia do.
Description
| Type of resource | text |
|---|---|
| Form | electronic resource; remote; computer; online resource |
| Extent | 1 online resource. |
| Place | California |
| Place | [Stanford, California] |
| Publisher | [Stanford University] |
| Copyright date | 2018; ©2018 |
| Publication date | 2018; 2018 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Bennett, Mariko Lynne |
|---|---|
| Degree supervisor | Barres, Ben |
| Degree supervisor | Shatz, Carla J |
| Thesis advisor | Barres, Ben |
| Thesis advisor | Shatz, Carla J |
| Thesis advisor | Palmer, Theo |
| Thesis advisor | Porteus, Matthew H |
| Thesis advisor | Wyss-Coray, Anton |
| Degree committee member | Palmer, Theo |
| Degree committee member | Porteus, Matthew H |
| Degree committee member | Wyss-Coray, Anton |
| Associated with | Stanford University, Neurosciences Program. |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Mariko Lynne Bennett. |
|---|---|
| Note | Submitted to the Neurosciences Program. |
| Thesis | Thesis Ph.D. Stanford University 2018. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2018 by Mariko L Bennett
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
Also listed in
Loading usage metrics...