Induction of human Somatostatin and Parvalbumin neurons by expressing a single transcription factor LIM Homeobox 6
- Nanjing Medical University, China
- ShanghaiTech University, China
- Fudan University, China
- Chinese Academy of Sciences, China
Abstract
Human GABAergic interneurons (GIN) are implicated in normal brain function as well as numerous mental disorders. However, generation of functional human GIN subtypes from human pluripotent stem cells (hPSCs) has not been established. By expressing LHX6, a transcriptional factor critical for GIN development, we have induced hPSCs to GINs, including somatostatin (SST, 29%) and parvalbumin (PV, 21%) neurons. Our RNAseq results also confirmed alteration of GINs identity with the overexpression of LHX6. Five months after transplantation into the mouse brain, the human GABA precursors generated increased population of SST and PV neurons by overexpressing LHX6. Importantly, the grafted human GINs exhibited functional electrophysiological properties and even fast-spiking like action potentials. Thus, expression of the single transcription factor LHX6 under our GIN differentiation condition is sufficient to robustly induce human PV and SST subtypes.
Data availability
Sequencing data have been deposited in GEO under accession codesGSE114553. All data generated or analysed during this study are included in the manuscript and supporting files.
-
Induction of human SST and PV neurons by expressing a single transcription factor LHX6Publicly available at the NCBI Gene Expression Omnibus (accession no: GSE114553).
Article and author information
Author details
Funding
National Natural Science Foundation of China (81471301)
- Yan Liu
National Key Research and Development Program of China (2016YFC1306703)
- Yan Liu
Jiangsu Outstanding Young Investigator Program (BK20160044)
- Yan Liu
ShanghaiTech Start-up Foundation
- Shuijin He
Postgraduate Research and Practice Innovation Program of Jiangsu Province
- Fang Yuan
Strategic Priority Research Program of the Chinese Academy of the Chinese Academy of Sciences (XDA16010306)
- Yan Liu
National Natural Science Foundation of China (31671063)
- Yan Liu
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: SCID mice were purchased from the Model Animal Research Center of Nanjing University. All of the animal experiments followed standard experimental protocols and were approved by the Animal Care and Use Committee at Nanjing Medical University(IACUC-1601129).
Human subjects: H9 cell line was purchased from Wicell Agreeement (NO.16-W0060)For ips cell line, verbal and written consent was obtained from the person.The study was approved by the ethic community of Nanjing Medical University[(2017）NO.217].
Copyright
© 2018, Yuan et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
Metrics
-
- 4,038
- views
-
- 610
- downloads
-
- 34
- citations
Views, downloads and citations are aggregated across all versions of this paper published by eLife.
Download links
A two-part list of links to download the article, or parts of the article, in various formats.
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
Induction of human Somatostatin and Parvalbumin neurons by expressing a single transcription factor LIM Homeobox 6
eLife 7:e37382.
https://doi.org/10.7554/eLife.37382
Further reading
-
- Cancer Biology
- Developmental Biology
Missense ‘hotspot’ mutations localized in six p53 codons account for 20% of TP53 mutations in human cancers. Hotspot p53 mutants have lost the tumor suppressive functions of the wildtype protein, but whether and how they may gain additional functions promoting tumorigenesis remain controversial. Here, we generated Trp53Y217C, a mouse model of the human hotspot mutant TP53Y220C. DNA damage responses were lost in Trp53Y217C/Y217C (Trp53YC/YC) cells, and Trp53YC/YC fibroblasts exhibited increased chromosome instability compared to Trp53-/- cells. Furthermore, Trp53YC/YC male mice died earlier than Trp53-/- males, with more aggressive thymic lymphomas. This correlated with an increased expression of inflammation-related genes in Trp53YC/YC thymic cells compared to Trp53-/- cells. Surprisingly, we recovered only one Trp53YC/YC female for 22 Trp53YC/YC males at weaning, a skewed distribution explained by a high frequency of Trp53YC/YC female embryos with exencephaly and the death of most Trp53YC/YC female neonates. Strikingly, however, when we treated pregnant females with the anti-inflammatory drug supformin (LCC-12), we observed a fivefold increase in the proportion of viable Trp53YC/YC weaned females in their progeny. Together, these data suggest that the p53Y217C mutation not only abrogates wildtype p53 functions but also promotes inflammation, with oncogenic effects in males and teratogenic effects in females.
-
- Developmental Biology
Stem cell self-renewal often relies on asymmetric fate determination governed by niche signals and/or cell-intrinsic factors but how these regulatory mechanisms cooperate to promote asymmetric fate decision remains poorly understood. In adult Drosophila midgut, asymmetric Notch (N) signaling inhibits intestinal stem cell (ISC) self-renewal by promoting ISC differentiation into enteroblast (EB). We have previously shown that epithelium-derived Bone Morphogenetic Protein (BMP) promotes ISC self-renewal by antagonizing N pathway activity (Tian and Jiang, 2014). Here, we show that loss of BMP signaling results in ectopic N pathway activity even when the N ligand Delta (Dl) is depleted, and that the N inhibitor Numb acts in parallel with BMP signaling to ensure a robust ISC self-renewal program. Although Numb is asymmetrically segregated in about 80% of dividing ISCs, its activity is largely dispensable for ISC fate determination under normal homeostasis. However, Numb becomes crucial for ISC self-renewal when BMP signaling is compromised. Whereas neither Mad RNA interference nor its hypomorphic mutation led to ISC loss, inactivation of Numb in these backgrounds resulted in stem cell loss due to precocious ISC-to-EB differentiation. Furthermore, we find that numb mutations resulted in stem cell loss during midgut regeneration in response to epithelial damage that causes fluctuation in BMP pathway activity, suggesting that the asymmetrical segregation of Numb into the future ISC may provide a fail-save mechanism for ISC self-renewal by offsetting BMP pathway fluctuation, which is important for ISC maintenance in regenerative guts.
