Molecular and Cellular Biochemistry

, Volume 450, Issue 1–2, pp 167–174 | Cite as

Loss of the basic helix-loop-helix transcription factor Bhlhe41 induces cell death and impairs neurite outgrowth in Neuro2a cells

  • Yajuan Sun
  • Haina Zhang
  • Libo Wang
  • Jia Li
  • Hui Jin
  • Zhenyu Wang
  • Shiyuan Tian
  • Ling Qi
  • Xiaoyang Liu


The basic helix-loop-helix (bHLH) superfamily of transcription factors have been implicated in a wide range of cellular functions such as proliferation, differentiation, tumorigenesis, and circadian rhythms. In a previous siRNA-based screen, bHLH family member e41 (BHLHE41) had been identified as a putative regulator of neuronal differentiation; however, its function remains largely elusive. To this end, using the CRISPR-Cas9 system, we established an isogenic Neuro2a (N2a) cell line with biallelic targeting of Bhlhe41 gene (Bhlhe41−/−). In undifferentiated N2a cells, complete knockout of Bhlhe41 resulted in marked proliferation inhibition, together with accumulation of apoptotic cells. Furthermore, retinoic acid (RA)-induced neurite outgrowth and expression of neuronal markers are significantly weakened in Bhlhe41−/− cells. We also showed that the activity of ERK1/2 signaling, a key regulator of neuronal differentiation, is likewise impaired in knockout cells. Together, these results suggest that Bhlhe41 plays critical roles in regulating cell death and neurite outgrowth in N2a cells.


Bhlhe41 Apoptosis Neurite outgrowth ERK1/2 signaling 



This study was supported by National Natural Science Foundation of China (81201671) and Foundation of Science and Technology Department of Jilin Province (20180101306JC, 20180101144JC).

Author contributions

XL, YS, and HZ designed the project, contributed to all experiments and to writing the manuscript. YS, HZ, LW, JL, and HJ conducted the experiments. ZW and ST helped with cell culture. LQ helped with data collection and analysis. All authors read and provided their approval for the final version of the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

The studies did not involve human participants or animals and therefore did not require ethical approval or informed consent to undertake.

Supplementary material

11010_2018_3383_MOESM1_ESM.pdf (69 kb)
Supplementary material 1 (PDF 69 KB)


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Yajuan Sun
    • 1
  • Haina Zhang
    • 2
  • Libo Wang
    • 1
  • Jia Li
    • 1
  • Hui Jin
    • 1
  • Zhenyu Wang
    • 1
  • Shiyuan Tian
    • 1
  • Ling Qi
    • 3
  • Xiaoyang Liu
    • 1
  1. 1.Department of NeurologyChina-Japan Union Hospital of Jilin UniversityChangchunPeople’s Republic of China
  2. 2.Department of RehabilitationThe Second Hospital of Jilin UniversityChangchunPeople’s Republic of China
  3. 3.Department of PathophysiologyJilin Medical UniversityJilinPeople’s Republic of China

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