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Expression of Neuropathy Target Esterase in mouse embryonic stem cells during differentiation

  • Organ Toxicity and Mechanisms
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Abstract

Neuropathy Target Esterase (NTE) was initially identified as the primary target esterase of some organophosphorus compounds that cause delayed neuropathy. Some studies in vivo suggest that this protein may also perform a function in embryonic development and therefore also in cell differentiation. The aim of this work was to characterize embryonic stem cells (ESC) as cellular model before to approach to the role of NTE in embryotoxicity processes through mechanistic studies. Mouse D3 ESC in monolayer expressed an NTE activity of 23 nmol phenol/min/mg of protein, while mouse R1 ESC showed a specific NTE activity 3 times higher than D3. An increased expression of gene Pnpla6 (that codifies for NTE) was seen during differentiation in both the D3 cells in monolayer and embryonic bodies (EBS). The maximums of the Pnpla6 expression were reached after 30 h and 5 days of differentiation in monolayer and EBS cultures, respectively. This peak of the Pnpla6 expression correlated with the peak of the NTE enzymatic activity in D3 monolayers. NTE activity and Pnpla6 expression returned to basal levels after 48 h (in monolayer cultures) and 10 days (in EBS) of differentiation, respectively. The changes in the Pnpla6 expression did not correlate with changes noted in the expression of two endoderm, two ectoderm and one neuroectoderm gene markers. In conclusion, this manuscript reports about NTE expression in ESC and its variation during first stages of differentiation. Nevertheless, the role of this activity and the meaning of the variations detected during differentiation must be further studied.

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Abbreviations

A-activity:

Total phenyl valerate esterase activity

Afp:

Alpha-fetoprotein

Amn:

Amnionless

B-activity:

Phenyl valerate esterase activity resistant to 40 µM paraoxon

C-activity:

Phenyl valerate esterase activity resistant to 40 µM paraoxon plus 250 µM mipafox

EBS:

Embryonic bodies

ESC:

Embryonic stem cells

LIF:

Leukemia inhibitory factor

MHC:

Myosin heavy chain

Nefm:

Intermediate neurofilament

NTE:

Neuropathy target esterase

PV:

Phenyl valerate

PVase:

Phenyl valerate esterase

qRT–PCR:

Quantitative real-time PCR, RT–PCR, conventional real-time PCR

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Acknowledgments

This study was supported by Spanish Ministry of the Environment (Grant A051/2007/3-14.4).

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Authors and Affiliations

  1. Unidad de Toxicología y seguridad Química, Instituto de Bioingeniería, Universidad Miguel Hernández de Elche, Avenida de la Universidad s/n, 03202, Elche, Spain

    David Pamies, Eugenio Vilanova & Miguel A. Sogorb

  2. Unidad de Bioquímica y Terapia Celular, Instituto de Bioingeniería, Universidad Miguel Hernández de Elche, Elche, Spain

    Juan Antonio Reig

Authors
  1. David Pamies
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  2. Juan Antonio Reig
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  3. Eugenio Vilanova
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  4. Miguel A. Sogorb
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Correspondence to Miguel A. Sogorb.

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Pamies, D., Reig, J.A., Vilanova, E. et al. Expression of Neuropathy Target Esterase in mouse embryonic stem cells during differentiation. Arch Toxicol 84, 481–491 (2010). https://doi.org/10.1007/s00204-010-0518-8

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  • DOI: https://doi.org/10.1007/s00204-010-0518-8

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