Hypoxia and hyperoxia differentially control proliferation of rat neural crest stem cells via distinct regulatory pathways of the HIF1α-CXCR4 and TP53-TPM1 proteins

Dev Dyn. 2017 Mar;246(3):162-185. doi: 10.1002/dvdy.24481. Epub 2017 Feb 3.

Abstract

Background: Neural crest stem cells (NCSCs) are a population of adult multipotent stem cells. We are interested in studying whether oxygen tensions affect the capability of NCSCs to self-renew and repair damaged tissues. NCSCs extracted from the hair follicle bulge region of the rat whisker pad were cultured in vitro under different oxygen tensions.

Results: We found significantly increased and decreased rates of cell proliferation in rat NCSCs (rNCSCs) cultured, respectively, at 0.5% and 80% oxygen levels. At 0.5% oxygen, the expression of both hypoxia-inducible factor (HIF) 1α and CXCR4 was greatly enhanced in the rNCSC nuclei and was suppressed by incubation with the CXCR4-specific antagonist AMD3100. In addition, the rate of cell apoptosis in the rNCSCs cultured at 80% oxygen was dramatically increased, associated with increased nuclear expression of TP53, decreased cytoplasmic expression of TPM1 (tropomyosin-1), and increased nuclear-to-cytoplasmic translocation of S100A2. Incubation of rNCSCs with the antioxidant N-acetylcysteine (NAC) overcame the inhibitory effect of 80% oxygen on proliferation and survival of rNCSCs.

Conclusions: Our results show for the first time that extreme oxygen tensions directly control NCSC proliferation differentially via distinct regulatory pathways of proteins, with hypoxia via the HIF1α-CXCR4 pathway and hyperoxia via the TP53-TPM1 pathway. Developmental Dynamics 246:162-185, 2017. © 2016 Wiley Periodicals, Inc.

Keywords: CDKN1A (p21CIP1/WAF1); CXCR4; HIF1α; S100A2; TP53; TPM1 (tropomyosin-1); antioxidant; hyperoxia; hypoxia; neural crest; oxygen; stem cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / genetics
  • Apoptosis / physiology
  • Blotting, Western
  • Cell Proliferation / genetics
  • Cell Proliferation / physiology
  • Cell Survival / genetics
  • Cell Survival / physiology
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / metabolism*
  • Female
  • Fluorescent Antibody Technique
  • Hair Follicle / cytology
  • Hair Follicle / metabolism
  • Hyperoxia / metabolism*
  • Hyperoxia / physiopathology
  • Hypoxia / metabolism*
  • Hypoxia / physiopathology
  • Hypoxia-Inducible Factor 1, alpha Subunit / genetics
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism*
  • Neural Crest / cytology*
  • Oxidative Stress / genetics
  • Oxidative Stress / physiology
  • RNA, Small Interfering / genetics
  • Rats
  • Rats, Sprague-Dawley
  • Receptors, CXCR4 / genetics
  • Receptors, CXCR4 / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction / genetics
  • Signal Transduction / physiology
  • Tropomyosin / genetics
  • Tropomyosin / metabolism*
  • Tumor Suppressor Protein p53 / genetics
  • Tumor Suppressor Protein p53 / metabolism*

Substances

  • Cxcr4 protein, rat
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • RNA, Small Interfering
  • Receptors, CXCR4
  • Tpm1 protein, rat
  • Tropomyosin
  • Tumor Suppressor Protein p53