The use of fluorescence for detecting MeHg-induced ROS in cell cultures

Parvinder Kaur, Kristina Schulz, Ingrid Heggland, Michael Aschner, Tore Syversen

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

The effect of methylmercury (MeHg) on reactive oxygen species (ROS) induction in neural cell lines was measured by the fluorescent probe, chloro methyl derivative of di-chloro di-hydro fluoresceindiacetate (CMH2DCFDA). Three different MeHg concentrations (5, 10 and 25 μM) and time periods (30, 50 and 90 min) were studied in C6-glial and B35-neuronal cell lines. In addition, the relationship between MeHg-induced ROS and cell density (day3 vs. day4) was also explored. The 14C-labelled MeHg measurements were done to determine the cell associated-MeHg content. At 30 and 50 min exposure, a significant increase (p < 0.05) in MeHg-induced ROS was observed at 10 and 25 μM MeHg for C6 cells and at 25 μM MeHg for B35 cells. However, the amount of ROS produced with 25 μM MeHg varied significantly (p < 0.001) at different time periods. For both the cell lines, significant cell density dependent differences (p < 0.05) were observed at 10 μM MeHg treatment for 50 min. MeHg treatments were associated with a concentration as well as cell-density dependent increase in cell associated-MeHg. These findings provide experimental evidence that special attention should be focused upon concentration, exposure time and cell density when assessing MeHg-induced ROS via fluorescence.

Original languageEnglish (US)
Pages (from-to)1392-1398
Number of pages7
JournalToxicology in Vitro
Volume22
Issue number5
DOIs
StatePublished - Aug 2008
Externally publishedYes

Fingerprint

Cell culture
Reactive Oxygen Species
Cell Culture Techniques
Fluorescence
Cell Count
Cells
Cell Line
Fluorescent Dyes
Neuroglia
Derivatives

Keywords

  • Fluorescence
  • In vitro
  • Methylmercury
  • Reactive oxygen species

ASJC Scopus subject areas

  • Toxicology

Cite this

The use of fluorescence for detecting MeHg-induced ROS in cell cultures. / Kaur, Parvinder; Schulz, Kristina; Heggland, Ingrid; Aschner, Michael; Syversen, Tore.

In: Toxicology in Vitro, Vol. 22, No. 5, 08.2008, p. 1392-1398.

Research output: Contribution to journalArticle

Kaur, Parvinder ; Schulz, Kristina ; Heggland, Ingrid ; Aschner, Michael ; Syversen, Tore. / The use of fluorescence for detecting MeHg-induced ROS in cell cultures. In: Toxicology in Vitro. 2008 ; Vol. 22, No. 5. pp. 1392-1398.
@article{a0f704c2c8e8471b83c0cc25d997c8d5,
title = "The use of fluorescence for detecting MeHg-induced ROS in cell cultures",
abstract = "The effect of methylmercury (MeHg) on reactive oxygen species (ROS) induction in neural cell lines was measured by the fluorescent probe, chloro methyl derivative of di-chloro di-hydro fluoresceindiacetate (CMH2DCFDA). Three different MeHg concentrations (5, 10 and 25 μM) and time periods (30, 50 and 90 min) were studied in C6-glial and B35-neuronal cell lines. In addition, the relationship between MeHg-induced ROS and cell density (day3 vs. day4) was also explored. The 14C-labelled MeHg measurements were done to determine the cell associated-MeHg content. At 30 and 50 min exposure, a significant increase (p < 0.05) in MeHg-induced ROS was observed at 10 and 25 μM MeHg for C6 cells and at 25 μM MeHg for B35 cells. However, the amount of ROS produced with 25 μM MeHg varied significantly (p < 0.001) at different time periods. For both the cell lines, significant cell density dependent differences (p < 0.05) were observed at 10 μM MeHg treatment for 50 min. MeHg treatments were associated with a concentration as well as cell-density dependent increase in cell associated-MeHg. These findings provide experimental evidence that special attention should be focused upon concentration, exposure time and cell density when assessing MeHg-induced ROS via fluorescence.",
keywords = "Fluorescence, In vitro, Methylmercury, Reactive oxygen species",
author = "Parvinder Kaur and Kristina Schulz and Ingrid Heggland and Michael Aschner and Tore Syversen",
year = "2008",
month = "8",
doi = "10.1016/j.tiv.2008.01.017",
language = "English (US)",
volume = "22",
pages = "1392--1398",
journal = "Toxicology in Vitro",
issn = "0887-2333",
publisher = "Elsevier Limited",
number = "5",

}

TY - JOUR

T1 - The use of fluorescence for detecting MeHg-induced ROS in cell cultures

AU - Kaur, Parvinder

AU - Schulz, Kristina

AU - Heggland, Ingrid

AU - Aschner, Michael

AU - Syversen, Tore

PY - 2008/8

Y1 - 2008/8

N2 - The effect of methylmercury (MeHg) on reactive oxygen species (ROS) induction in neural cell lines was measured by the fluorescent probe, chloro methyl derivative of di-chloro di-hydro fluoresceindiacetate (CMH2DCFDA). Three different MeHg concentrations (5, 10 and 25 μM) and time periods (30, 50 and 90 min) were studied in C6-glial and B35-neuronal cell lines. In addition, the relationship between MeHg-induced ROS and cell density (day3 vs. day4) was also explored. The 14C-labelled MeHg measurements were done to determine the cell associated-MeHg content. At 30 and 50 min exposure, a significant increase (p < 0.05) in MeHg-induced ROS was observed at 10 and 25 μM MeHg for C6 cells and at 25 μM MeHg for B35 cells. However, the amount of ROS produced with 25 μM MeHg varied significantly (p < 0.001) at different time periods. For both the cell lines, significant cell density dependent differences (p < 0.05) were observed at 10 μM MeHg treatment for 50 min. MeHg treatments were associated with a concentration as well as cell-density dependent increase in cell associated-MeHg. These findings provide experimental evidence that special attention should be focused upon concentration, exposure time and cell density when assessing MeHg-induced ROS via fluorescence.

AB - The effect of methylmercury (MeHg) on reactive oxygen species (ROS) induction in neural cell lines was measured by the fluorescent probe, chloro methyl derivative of di-chloro di-hydro fluoresceindiacetate (CMH2DCFDA). Three different MeHg concentrations (5, 10 and 25 μM) and time periods (30, 50 and 90 min) were studied in C6-glial and B35-neuronal cell lines. In addition, the relationship between MeHg-induced ROS and cell density (day3 vs. day4) was also explored. The 14C-labelled MeHg measurements were done to determine the cell associated-MeHg content. At 30 and 50 min exposure, a significant increase (p < 0.05) in MeHg-induced ROS was observed at 10 and 25 μM MeHg for C6 cells and at 25 μM MeHg for B35 cells. However, the amount of ROS produced with 25 μM MeHg varied significantly (p < 0.001) at different time periods. For both the cell lines, significant cell density dependent differences (p < 0.05) were observed at 10 μM MeHg treatment for 50 min. MeHg treatments were associated with a concentration as well as cell-density dependent increase in cell associated-MeHg. These findings provide experimental evidence that special attention should be focused upon concentration, exposure time and cell density when assessing MeHg-induced ROS via fluorescence.

KW - Fluorescence

KW - In vitro

KW - Methylmercury

KW - Reactive oxygen species

UR - http://www.scopus.com/inward/record.url?scp=44749084245&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=44749084245&partnerID=8YFLogxK

U2 - 10.1016/j.tiv.2008.01.017

DO - 10.1016/j.tiv.2008.01.017

M3 - Article

VL - 22

SP - 1392

EP - 1398

JO - Toxicology in Vitro

JF - Toxicology in Vitro

SN - 0887-2333

IS - 5

ER -