Transcripts involved in calcium signaling and telencephalic neuronal fate are altered in induced pluripotent stem cells from bipolar disorder patients.
paper
Cited
Public
- Authors
- Chen, H M; DeLong, C J; Bame, M; Rajapakse, I; Herron, T J; McInnis, M G; O'Shea, K S
- Year
- 2014
- Journal
- Translational psychiatry
- PMID
- 25116795
- DOI
- 10.1038/tp.2014.12
- PMCID
- PMC3966040
Bipolar disorder (BP) is a chronic psychiatric condition characterized by dynamic, pathological mood fluctuations from mania to depression. To date, a major challenge in studying human neuropsychiatric conditions such as BP has been limited access to viable central nervous system tissue to examine disease progression. Patient-derived induced pluripotent stem cells (iPSCs) now offer an opportunity to analyze the full compliment of neural tissues and the prospect of identifying novel disease mechanisms. We have examined changes in gene expression as iPSC derived from well-characterized patients differentiate into neurons; there was little difference in the transcriptome of iPSC, but BP neurons were significantly different than controls in their transcriptional profile. Expression of transcripts for membrane bound receptors and ion channels was significantly increased in BP-derived neurons compared with controls, and we found that lithium pretreatment of BP neurons significantly altered their calcium transient and wave amplitude. The expression of transcription factors involved in the specification of telencephalic neuronal identity was also altered. Control neurons expressed transcripts that confer dorsal telencephalic fate, whereas BP neurons expressed genes involved in the differentiation of ventral (medial ganglionic eminence) regions. Cells were responsive to dorsal/ventral patterning cues, as addition of the Hedgehog (ventral) pathway activator purmorphamine or a dorsalizing agent (lithium) stimulated expression of NKX2-1 (ventral identity) or EMX2 (dorsal) in both groups. Cell-based models should have a significant impact on our understanding of the genesis and therefore treatment of BP; the iPSC cell lines themselves provide an important resource for comparison with other neurodevelopmental disorders.
Derivation, characterization and neuronal differentiation of iPSC. Fibroblasts (a) express Te-7 (b; Cy3-secondary antibody, red). Reprogrammed colonies of iPSC form and are passaged to MEFs. They downregulate Te-7 (c; Cy3-secondary antibody), and exhibit nuclear expression of Nanog (d; FITC-secondary antibody). For differentiation, cells are removed from MEF and grown in defined medium as neurospheres (e), then plated to form rosettes (f). Rosettes are picked twice, then differentiated in adherent culture in defined medium forming nestin + (g; FITC-secondary antibody) neural progenitor cells, and with additional time in vitro undergo widespread differentiation to Ξ²-III tubulin + neurons (h; Cy3-secondary antibody), with punctate expression of Synapsin 1 (Syn1) along their processes (i, Cy3-secondary antibody). All cells, but a, are stained with Hoechst 23487 (blue) to identify nuclei. Scale bars in aβd,g = 500 mm; e = 2 mm, f,h,i = 200 mm.
LLM interpretation
This figure consists of a series of microscopy images (brightfield and immunofluorescence) illustrating the reprogramming of fibroblasts into iPSCs and their subsequent differentiation into neurons. Panels (a-d) show the transition from Te-7 positive fibroblasts to Nanog-expressing iPSC colonies, while panels (e-f) show the formation of neurospheres and rosettes. Panels (g-i) demonstrate the final stages of differentiation, showing the expression of Nestin in neural progenitor cells, $\beta$-III tubulin in neurons, and punctate Synapsin 1 (Syn1) along neuronal processes.
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