Altered temporal sequence of transcriptional regulators in the generation of human cerebellar granule cells
- Rockefeller University, United States
Abstract
Brain development is regulated by conserved transcriptional programs across species, but little is known about divergent mechanisms that create species-specific characteristics. Among brain regions, human cerebellar histogenesis differs in complexity compared with non-human primates and rodents, making it important to develop methods to generate human cerebellar neurons that closely resemble those in the developing human cerebellum. We report a rapid protocol for the derivation of the human ATOH1 lineage, the precursor of excitatory cerebellar neurons, from human pluripotent stem cells (hPSC). Upon transplantation into juvenile mice, hPSC-derived cerebellar granule cells migrated along glial fibers and integrated into the cerebellar cortex. By Translational Ribosome Affinity Purification-seq, we identified an unexpected temporal shift in the expression of RBFOX3 (NeuN) and NEUROD1, which are classically associated with differentiated neurons, in the human outer external granule layer. This molecular divergence may enable the protracted development of the human cerebellum compared to mice.
Data availability
Sequencing data have been deposited in GEO under accession code: GSE163710. For reviewers only, a temporary password has been generated: ejkpqqeupdkplcx.Upon publication, the data will be released publicly.
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TRAP seq of the human pluripotent stem cell derived ATOH1 lineageNBCI Gene Expression Omnibus, GSE163710.
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Integrative functional genomic analysis of human brain development and neuropsychiatric risksHuman mRNA seq processed data: Gene expression in counts.
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Specification of diverse cell types during early neurogenesis of the mouse cerebellumhttps://doi.org/10.7554/eLife.42388.018.
Article and author information
Author details
Funding
National Institute of Neurological Disorders and Stroke (1R21NS093540-01)
- Mary E Hatten
The Rockefeller University Center for Clinical and Translational Science (Pilot award)
- Hourinaz Behesti
- Mary E Hatten
Starr Foundation (Tri-Institutional Stem Cell Initiative Grant)
- Mary E Hatten
Department of Defense US Army Medical Research Acquisition Activity Grants (W81XWH1510189)
- Mary E Hatten
The Robertson Therapeutics Development Fund
- Mary E Hatten
Renate, Hans, and Maria Hofmann Trust
- Mary E Hatten
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Carol A Mason, Columbia University, United States
Ethics
Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocol (#14746-H) of the Rockefeller University. All surgery was performed under hypothermia, and every effort was made to minimize suffering.
Human subjects: Fixed de-identified human tissue were acquired from the Human Developmental Biology Resource (http://www.hdbr.org/) following institutional policies.
Version history
- Preprint posted: January 17, 2021 (view preprint)
- Received: January 30, 2021
- Accepted: November 27, 2021
- Accepted Manuscript published: November 29, 2021 (version 1)
- Version of Record published: December 20, 2021 (version 2)
Copyright
© 2021, Behesti 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.
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Altered temporal sequence of transcriptional regulators in the generation of human cerebellar granule cells
eLife 10:e67074.
https://doi.org/10.7554/eLife.67074
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