Electrical activity and cell fate specification in the normal developing brain and in neuropsychiatric disease
Abstract/Contents
- Abstract
- Mutations in CACNA1C, the gene encoding the L-type voltage gated calcium channel (LTC) Cav1.2, have been associated with autism, schizophrenia, and bipolar disorder. In this thesis, I investigated the role of activity through the LTC Cav1.2 on the generation of excitatory projection neurons in the developing cortex. LTCs such as Cav1.2 convert electrical activity into calcium signals that activate programs of gene expression in the developing nervous system. A gain of function mutation in an alternatively spliced exon of Cav1.2 causes Timothy Syndrome (TS), a multisystemic disorder characterized by cardiac arrhythmias and autism. Using an induced pluripotent stem cell (iPSC) platform, our lab previously reported that neurons from TS patients have altered gene expression suggesting a change in the abundance and laminar identity of early-born cortical projection neurons. As part of my thesis work, I demonstrated that Cav1.2 is expressed in the developing cortex in the mouse and human brain, and that GABA depolarization-induced calcium rises in NPCs and immature neurons can be completely blocked by pharmacological inhibitors of LTCs. I then found that splicing of Cav1.2 is dynamically regulated during mouse embryonic brain development and in human iPSC-derived cells. The exon containing the TS mutation is predominantly expressed in immature cells, and the TS mutation causes subtle alterations in Cav1.2 mRNA splicing, resulting in overabundance of the mutant exon in Cav1.2 transcripts. Through a series of in utero electroporation experiments mimicking TS, I found that over-expressing TS or wild type Cav1.2 in vivo recapitulates the differentiation defects reflected by our gene expression studies in iPSC-derived TS neurons, resulting in reduced SATB2-expressing putative callosally projecting neurons and increased CTIP2-expressing subcortically projecting cells. Over-expressing a channel that cannot carry calcium eliminates this effect, supporting the idea that excess calcium signaling may underlie differentiation defects observed in TS patient cells. In utero loss of function of Cav1.2 has the opposite effect, resulting in an overabundance of SATB2-expressing cells in the cortical plate. In a collaborative project exploring another genetically defined form of autism caused by deletion of the 16p11.2 genomic locus, we also observed a reduction in SATB2-expressing cells, suggesting the possibility of common cortical differentiation defects across ASDs. Together, this work indicates that altered expression of Cav1.2 can bidirectionally regulate the differentiation of early-born cortical projection neurons and seeds the idea that the abundance of SATB2 and CTIP2-expressing cells may be a point of convergence for multiple psychiatric disorders.
Description
| Type of resource | text |
|---|---|
| Form | electronic; electronic resource; remote |
| Extent | 1 online resource. |
| Publication date | 2014 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Associated with | Panagiotakos, Georgia | |
|---|---|---|
| Associated with | Stanford University, Neurosciences Program. | |
| Primary advisor | Dolmetsch, Ricardo E | |
| Primary advisor | Palmer, Theo | |
| Thesis advisor | Dolmetsch, Ricardo E | |
| Thesis advisor | Palmer, Theo | |
| Thesis advisor | Clandinin, Thomas R. (Thomas Robert), 1970- | |
| Thesis advisor | McConnell, Susan K | |
| Advisor | Clandinin, Thomas R. (Thomas Robert), 1970- | |
| Advisor | McConnell, Susan K |
Subjects
| Genre | Theses |
|---|
Bibliographic information
| Statement of responsibility | Georgia Panagiotakos. |
|---|---|
| Note | Submitted to the Program in Neurosciences. |
| Thesis | Thesis (Ph.D.)--Stanford University, 2014. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2014 by Georgia Panagiotakos
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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