Critical roles of Src family tyrosine kinases in excitatory neuronal differentiation of cultured embryonic stem cells

Michelle Hedrick Theus, Ling Wei, Kevin Francis, Shan Ping Yu

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Embryonic stem (ES) cells have been tested for potential cell transplantation therapy for CNS disorders. Understanding their differentiation mechanism and identifying factors involved in driving excitatory and inhibitory neuron lineages should enhance the efficacy and efficiency of the cell transplantation therapy. We tested the hypothesis that selective expression of Src family tyrosine kinases is required for phenotype-specific differentiation and functional maturation of ES cell derived neurons. Cultured mouse pluripotent ES cells were treated with retinoic acid (RA) to induce neural differentiation. After RA induction, neurons derived from ES cells showed significant neurite growth, increased expression of Src, Fyn and Lck and an extension of Src kinase expression from cell body to neurite processes. ES cell derived neuron-like cells expressed neurofilament, synaptophysin, glutamate receptors, NMDA and kainate currents, became vulnerable to excitotoxicity and formed functional excitatory synapses. These developmental events were blocked or attenuated when cells were grown in the presence of Src family kinase inhibitor PP2. However, there was no change in the expression of GABAergic-specific protein GAD67 during PP2 treatment. Our data suggest that Src tyrosine kinases are involved in the terminal differentiation of excitatory neuronal phenotype during ES cell neural differentiation after RA induction.

Original languageEnglish (US)
Pages (from-to)3096-3107
Number of pages12
JournalExperimental Cell Research
Volume312
Issue number16
DOIs
StatePublished - Oct 1 2006

Fingerprint

src-Family Kinases
Embryonic Stem Cells
Tretinoin
Neurons
Cell Transplantation
Neurites
Cell- and Tissue-Based Therapy
Phenotype
Pluripotent Stem Cells
Synaptophysin
Intermediate Filaments
Kainic Acid
Glutamate Receptors
N-Methylaspartate
Synapses
Cell Differentiation
Growth
Proteins

Keywords

  • Differentiation
  • Embryonic stem cells
  • GABAergic neurons
  • Glutamatergic neurons
  • NMDA receptor
  • Neurite outgrowth
  • Src kinases
  • mEPSC

ASJC Scopus subject areas

  • Cell Biology

Cite this

Critical roles of Src family tyrosine kinases in excitatory neuronal differentiation of cultured embryonic stem cells. / Hedrick Theus, Michelle; Wei, Ling; Francis, Kevin; Ping Yu, Shan.

In: Experimental Cell Research, Vol. 312, No. 16, 01.10.2006, p. 3096-3107.

Research output: Contribution to journalArticle

@article{050f0b0454ce4970bdc48052bf480483,
title = "Critical roles of Src family tyrosine kinases in excitatory neuronal differentiation of cultured embryonic stem cells",
abstract = "Embryonic stem (ES) cells have been tested for potential cell transplantation therapy for CNS disorders. Understanding their differentiation mechanism and identifying factors involved in driving excitatory and inhibitory neuron lineages should enhance the efficacy and efficiency of the cell transplantation therapy. We tested the hypothesis that selective expression of Src family tyrosine kinases is required for phenotype-specific differentiation and functional maturation of ES cell derived neurons. Cultured mouse pluripotent ES cells were treated with retinoic acid (RA) to induce neural differentiation. After RA induction, neurons derived from ES cells showed significant neurite growth, increased expression of Src, Fyn and Lck and an extension of Src kinase expression from cell body to neurite processes. ES cell derived neuron-like cells expressed neurofilament, synaptophysin, glutamate receptors, NMDA and kainate currents, became vulnerable to excitotoxicity and formed functional excitatory synapses. These developmental events were blocked or attenuated when cells were grown in the presence of Src family kinase inhibitor PP2. However, there was no change in the expression of GABAergic-specific protein GAD67 during PP2 treatment. Our data suggest that Src tyrosine kinases are involved in the terminal differentiation of excitatory neuronal phenotype during ES cell neural differentiation after RA induction.",
keywords = "Differentiation, Embryonic stem cells, GABAergic neurons, Glutamatergic neurons, NMDA receptor, Neurite outgrowth, Src kinases, mEPSC",
author = "{Hedrick Theus}, Michelle and Ling Wei and Kevin Francis and {Ping Yu}, Shan",
year = "2006",
month = "10",
day = "1",
doi = "10.1016/j.yexcr.2006.06.022",
language = "English (US)",
volume = "312",
pages = "3096--3107",
journal = "Experimental Cell Research",
issn = "0014-4827",
publisher = "Academic Press Inc.",
number = "16",

}

TY - JOUR

T1 - Critical roles of Src family tyrosine kinases in excitatory neuronal differentiation of cultured embryonic stem cells

AU - Hedrick Theus, Michelle

AU - Wei, Ling

AU - Francis, Kevin

AU - Ping Yu, Shan

PY - 2006/10/1

Y1 - 2006/10/1

N2 - Embryonic stem (ES) cells have been tested for potential cell transplantation therapy for CNS disorders. Understanding their differentiation mechanism and identifying factors involved in driving excitatory and inhibitory neuron lineages should enhance the efficacy and efficiency of the cell transplantation therapy. We tested the hypothesis that selective expression of Src family tyrosine kinases is required for phenotype-specific differentiation and functional maturation of ES cell derived neurons. Cultured mouse pluripotent ES cells were treated with retinoic acid (RA) to induce neural differentiation. After RA induction, neurons derived from ES cells showed significant neurite growth, increased expression of Src, Fyn and Lck and an extension of Src kinase expression from cell body to neurite processes. ES cell derived neuron-like cells expressed neurofilament, synaptophysin, glutamate receptors, NMDA and kainate currents, became vulnerable to excitotoxicity and formed functional excitatory synapses. These developmental events were blocked or attenuated when cells were grown in the presence of Src family kinase inhibitor PP2. However, there was no change in the expression of GABAergic-specific protein GAD67 during PP2 treatment. Our data suggest that Src tyrosine kinases are involved in the terminal differentiation of excitatory neuronal phenotype during ES cell neural differentiation after RA induction.

AB - Embryonic stem (ES) cells have been tested for potential cell transplantation therapy for CNS disorders. Understanding their differentiation mechanism and identifying factors involved in driving excitatory and inhibitory neuron lineages should enhance the efficacy and efficiency of the cell transplantation therapy. We tested the hypothesis that selective expression of Src family tyrosine kinases is required for phenotype-specific differentiation and functional maturation of ES cell derived neurons. Cultured mouse pluripotent ES cells were treated with retinoic acid (RA) to induce neural differentiation. After RA induction, neurons derived from ES cells showed significant neurite growth, increased expression of Src, Fyn and Lck and an extension of Src kinase expression from cell body to neurite processes. ES cell derived neuron-like cells expressed neurofilament, synaptophysin, glutamate receptors, NMDA and kainate currents, became vulnerable to excitotoxicity and formed functional excitatory synapses. These developmental events were blocked or attenuated when cells were grown in the presence of Src family kinase inhibitor PP2. However, there was no change in the expression of GABAergic-specific protein GAD67 during PP2 treatment. Our data suggest that Src tyrosine kinases are involved in the terminal differentiation of excitatory neuronal phenotype during ES cell neural differentiation after RA induction.

KW - Differentiation

KW - Embryonic stem cells

KW - GABAergic neurons

KW - Glutamatergic neurons

KW - NMDA receptor

KW - Neurite outgrowth

KW - Src kinases

KW - mEPSC

UR - http://www.scopus.com/inward/record.url?scp=33747884882&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33747884882&partnerID=8YFLogxK

U2 - 10.1016/j.yexcr.2006.06.022

DO - 10.1016/j.yexcr.2006.06.022

M3 - Article

C2 - 16859680

AN - SCOPUS:33747884882

VL - 312

SP - 3096

EP - 3107

JO - Experimental Cell Research

JF - Experimental Cell Research

SN - 0014-4827

IS - 16

ER -