Neurochemical Research

, Volume 41, Issue 1–2, pp 5–15 | Cite as

Upregulation of Slc38a1 Gene Along with Promotion of Neurosphere Growth and Subsequent Neuronal Specification in Undifferentiated Neural Progenitor Cells Exposed to Theanine

  • Takeshi Takarada
  • Masato Ogura
  • Noritaka Nakamichi
  • Takami Kakuda
  • Ryota Nakazato
  • Hiroshi Kokubo
  • Shinsuke Ikeno
  • Saki Nakamura
  • Takaya Kutsukake
  • Eiichi Hinoi
  • Yukio Yoneda
Original Paper

Abstract

We have shown marked promotion of both cluster growth and neuronal specification in pluripotent P19 cells with overexpression of solute carrier 38a1 (Slc38a1), which is responsible for membrane transport of glutamine. In this study, we evaluated pharmacological profiles of the green tea amino acid ingredient theanine, which is a good substrate for glutamine transporters, on proliferation and neuronal specification in neural progenitor cells from embryonic rat neocortex. Sustained exposure to theanine, but not glutamine, accelerated the growth of neurospheres composed of proliferating cells and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) reducing activity at concentrations of 1–100 μM in undifferentiated progenitor cells. Such prior exposure to theanine promoted spontaneous and induced commitment to a neuronal lineage with concomitant deteriorated astroglial specification. Selective upregulation was seen in the expression of Slc38a1 in progenitor cells cultured with theanine. Similarly significant increases in cluster growth and MTT reducing activity were found in P19 cells cultured with theanine for 4 days. Luciferase activity was doubled in a manner sensitive to the deletion of promoter regions in P19 cells with a luciferase reporter plasmid of the Slc38a1 promoter after sustained exposure to theanine for 4 days. Overexpression of X-box binding protein-1 led to a marked increase in luciferase activity in P19 cells transfected with the Slc38a1 reporter plasmid. These results suggest that theanine accelerates cellular proliferation and subsequent neuronal specification through a mechanism relevant to upregulation of Slc38a1 gene in undifferentiated neural progenitor cells.

Keywords

Theanine Glutamine transporter SLC38A1 Neural progenitors Neurosphere Neuronal specification 

Abbreviations

AP-1

Activator protein-1

ATF2

Activating transcription factor-2

ATF2

Activating transcription factor-2

ATF3

Activating transcription factor-3

ATRA

All-trans retinoic acid

bHLH

Basic helix-loop-helix

BrdU

5-Bromo-2′-deoxyuridine

CaATF6

Constitutive active activating transcription factor-6

CaHIF1α

Constitutive active hypoxia inducible factor-1α

C/EBPα

CCAAT enhancer binding protein-α

CNTF

Ciliary neurotrophic factor

CNTFRα

Ciliary neurotrophic factor receptor alpha

CREB

Cyclic AMP responsive element binding protein

DMEM/F-12

Dulbecco’s modified Eagle medium: Nutrient Mixture F-12

ER

Endoplasmic reticulum

EV

Empty vector

FBS

Fetal bovine serum

FGF

Fibroblast growth factor

GAPDH

Glyceraldehyde-3-phosphate dehydrogenase

GFAP

Glial fibrillary acidic protein

Gln

Glutamine

HIF1α

Hypoxia inducible factor-1α

MAP2

Microtubules-associated protein-2

MTT

3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide

NCAM

Neural cell adhesion molecule

NFAT2

Nuclear factor of activated T cell-1

NF-κB

Nuclear factor-κB

Nrf2

Nuclear factor (erythroid-derived 2)-like 2

PA

Paraformaldehyde

PBS

Phosphate-buffered saline

PCR

Polymerase chain reaction

RARα

Retinoic acid receptor alpha

RT-PCR

Reverse transcription polymerase chain reaction

SLC38A1

Solute carrier 38a1

STAT1

Signal transducer and activator of transcription-1

STAT3

Signal transducer and activator of transcription-3

WT

Wild-type

XBP1

X-box binding protein-1

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Takeshi Takarada
    • 1
  • Masato Ogura
    • 1
    • 3
  • Noritaka Nakamichi
    • 1
    • 4
  • Takami Kakuda
    • 2
  • Ryota Nakazato
    • 1
  • Hiroshi Kokubo
    • 1
  • Shinsuke Ikeno
    • 1
  • Saki Nakamura
    • 1
  • Takaya Kutsukake
    • 1
  • Eiichi Hinoi
    • 1
  • Yukio Yoneda
    • 1
  1. 1.Laboratory of Molecular Pharmacology, Division of Pharmaceutical SciencesKanazawa University Graduate School of Medical, Pharmaceutical and Health SciencesKakuma-machi, KanazawaJapan
  2. 2.Research Center of Composite MaterialFukuoka UniversityFukuokaJapan
  3. 3.Department of Biomolecular ScienceFukushima Medical University School of MedicineFukushimaJapan
  4. 4.Laboratory of Molecular PharmacotherapeuticsKanazawa University Graduate SchoolKanazawaJapan

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