Altered temporal sequence of transcriptional regulators in the generation of human cerebellar granule cells

Abstract
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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.

The following data sets were generated

(2020) TRAP seq of the human pluripotent stem cell derived ATOH1 lineage
NBCI Gene Expression Omnibus, GSE163710.

https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE163710

The following previously published data sets were used

1. Mingfeng Li et al
(2018) Integrative functional genomic analysis of human brain development and neuropsychiatric risks
Human mRNA seq processed data: Gene expression in counts.

http://development.psychencode.org
(2019) Specification of diverse cell types during early neurogenesis of the mouse cerebellum
https://doi.org/10.7554/eLife.42388.018.

https://elifesciences.org/articles/42388

Article and author information

Author details

Hourinaz Behesti

Laboratory of Developmental Neurobiology, Rockefeller University, New York, United States

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0001-9383-9929
Arif Kocabas

Laboratory of Developmental Neurobiology, Rockefeller University, New York, United States

Competing interests
No competing interests declared.
David E Buchholz

Laboratory of Developmental Neurobiology, Rockefeller University, New York, United States

Competing interests
No competing interests declared.
Thomas S Carroll

Bioinformatics Resouce Center, Rockefeller University, New York, United States

Competing interests
No competing interests declared.
Mary E Hatten

Laboratory of Developmental Neurobiology, Rockefeller University, New York, United States

For correspondence
hatten@rockefeller.edu

Competing interests
Mary E Hatten, Reviewing editor, eLife.

"This ORCID iD identifies the author of this article:" 0000-0001-9059-660X

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.

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.

Copyright

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.