Loss of ZC4H2 and RNF220 Inhibits Neural Stem Cell Proliferation and Promotes Neuronal Differentiation

Longlong Zhang; Maosen Ye; Liang Zhu; Jingmei Cha; Chaocui Li; Yong-Gang Yao; Bingyu Mao

doi:10.3390/cells9071600

Loss of ZC4H2 and RNF220 Inhibits Neural Stem Cell Proliferation and Promotes Neuronal Differentiation

Cells. 2020 Jul 1;9(7):1600. doi: 10.3390/cells9071600.

Authors

Longlong Zhang^{1

2}, Maosen Ye^{2

3}, Liang Zhu^{1

2}, Jingmei Cha^{1

2}, Chaocui Li¹, Yong-Gang Yao^{2

3

4}, Bingyu Mao^{1

5}

Affiliations

¹ State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China.
² Kunming College of Life Science, University of Chinese Academy of Sciences, Kunming 650204, China.
³ Key Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences and Yunnan Province, and KIZ-CUHK Joint Laboratory of Bioresources and Molecular Research in Common Diseases, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China.
⁴ CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai 200031, China.
⁵ Center for Excellence in Animal Evolution and Genetics, Chinese Academy of Sciences, Kunming, Yunnan 650223, China.

Abstract

The ubiquitin E3 ligase RNF220 and its co-factor ZC4H2 are required for multiple neural developmental processes through different targets, including spinal cord patterning and the development of the cerebellum and the locus coeruleus. Here, we explored the effects of loss of ZC4H2 and RNF220 on the proliferation and differentiation of neural stem cells (NSCs) derived from mouse embryonic cortex. We showed that loss of either ZC4H2 or RNF220 inhibits the proliferation and promotes the differentiation abilities of NSCs in vitro. RNA-Seq profiling revealed 132 and 433 differentially expressed genes in the ZCH2^-/- and RNF220^-/- NSCs, compared to wild type (WT) NSCs, respectively. Specifically, Cend1, a key regulator of cell cycle exit and differentiation of neuronal precursors, was found to be upregulated in both ZCH2^-/- and RNF220^-/- NSCs at the mRNA and protein levels. The targets of Cend1, such as CyclinD1, Notch1 and Hes1, were downregulated both in ZCH2^-/- and RNF220^-/- NSCs, whereas p53 and p21 were elevated. ZCH2^-/- and RNF220^-/- NSCs showed G0/G1 phase arrest compared to WT NSCs in cell cycle analysis. These results suggested that ZC4H2 and RNF220 are likely involved in the regulation of neural stem cell proliferation and differentiation through Cend1.

Keywords: Cend1; RNF220; ZC4H2; differentiation; neural stem cell; proliferation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Proliferation / genetics*
Cerebellar Cortex / cytology
Cerebellar Cortex / metabolism
Cyclin D1 / genetics
Cyclin D1 / metabolism
Cyclin-Dependent Kinase Inhibitor p21 / genetics
Cyclin-Dependent Kinase Inhibitor p21 / metabolism
Down-Regulation
G1 Phase Cell Cycle Checkpoints / genetics
Intracellular Signaling Peptides and Proteins / genetics
Intracellular Signaling Peptides and Proteins / metabolism*
Membrane Proteins / genetics
Membrane Proteins / metabolism
Mice
Mice, Knockout
Nerve Tissue Proteins / genetics
Nerve Tissue Proteins / metabolism
Neural Stem Cells / cytology
Neural Stem Cells / metabolism*
Neurogenesis / genetics*
Nuclear Proteins / genetics
Nuclear Proteins / metabolism*
RNA-Seq
Receptor, Notch1 / genetics
Receptor, Notch1 / metabolism
Transcription Factor HES-1 / genetics
Transcription Factor HES-1 / metabolism
Tumor Suppressor Protein p53 / genetics
Tumor Suppressor Protein p53 / metabolism
Ubiquitin-Protein Ligases / genetics
Ubiquitin-Protein Ligases / metabolism*
Up-Regulation

Substances

Ccnd1 protein, mouse
Cend1 protein, mouse
Cyclin-Dependent Kinase Inhibitor p21
Hes1 protein, mouse
Intracellular Signaling Peptides and Proteins
Membrane Proteins
Nerve Tissue Proteins
Notch1 protein, mouse
Nuclear Proteins
Receptor, Notch1
Transcription Factor HES-1
Trp53 protein, mouse
Tumor Suppressor Protein p53
Zc4h2 protein, mouse
Cyclin D1
RNF220 protein, mouse
Ubiquitin-Protein Ligases