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Markers associated with neuron-specific Ube3a imprinting during neuronal differentiation of mouse embryonic stem cells

Authors
  • Eitoku, Masamitsu1
  • Kato, Hidemasa2
  • Suganuma, Narufumi1
  • Kiyosawa, Hidenori1, 3
  • 1 Kochi University, Department of Environmental Medicine, Kochi Medical School, Oko-Cho Kohasu Nankoku, Kochi, 783-8505, Japan , Oko-Cho Kohasu Nankoku (Japan)
  • 2 Ehime University, Division of Functional Histology, Department of Functional Biomedicine, Graduate School of Medicine, Shitsukawa, Toon, Ehime, 791-0295, Japan , Toon (Japan)
  • 3 Chiba Institute of Technology, Department of Life Science, Tsudanuma, Narashino, Chiba, 275-0016, Japan , Narashino, Chiba (Japan)
Type
Published Article
Journal
Cytotechnology
Publisher
Springer-Verlag
Publication Date
Aug 05, 2017
Volume
70
Issue
1
Pages
45–53
Identifiers
DOI: 10.1007/s10616-017-0126-z
Source
Springer Nature
Keywords
License
Yellow

Abstract

Understanding gene expression in the brain requires allele-specific transcriptome analysis because of the presence of neuron-specific imprinted genes, which are expressed in a neuron-specific and parent-of-origin-specific manner. Ube3a is a neuron-specific imprinted gene with an expression pattern that changes from biallelic to maternal only (Ube3a imprinting) during differentiation. Because Ube3a imprinting occurs only in neurons, it has the potential to be a marker to assess the quality of neurons produced by in vitro neuronal differentiation of embryonic stem cells (ESCs). For the analysis of Ube3a imprinting, genetic polymorphisms between the two alleles are necessary to identify the parental origin of each. However, ESCs derived from commonly used inbred mouse strains have no genetic polymorphisms. To overcome this problem, we examined 10 markers of neurogenesis to determine whether they were associated with Ube3a imprinting. We measured the relative expression levels of these 10 gene markers and assessed the Ube3a imprinting status of 54 neuron samples differentiated under various in vitro conditions. Then we divided the samples into two groups depending on their Ube3a imprinting status and selected markers statistically associated with Ube3a imprinting. The identified markers included the antisense noncoding transcript of Ube3a and a mature neuron marker Mtap2, consistent with the markers we used empirically in our previous study to assess the quality of differentiated neurons. These findings provide new quality control criteria for differentiated neurons, and could also be applied to human ESCs and induced pluripotent stem cells.

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