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Prion Protein Does Not Confer Resistance to Hippocampus-Derived Zpl Cells against the Toxic Effects of Cu2+, Mn2+, Zn2+ and Co2+ Not Supporting a General Protective Role for PrP in Transition Metal Induced Toxicity.

Cingaram PK, Nyeste A, Dondapati DT, Fodor E, Welker E - PLoS ONE (2015)

Bottom Line: By employing a cell viability assay, we examined the effects of various concentrations of Cu2+, Zn2+, Mn2+, and Co2+ on Zpl (Prnp-/-) and ZW (Prnp+/+) hippocampus-derived mouse neuronal cells.However, when we introduced PrP or only the empty vector into Zpl cells, we could not discern any protective effect associated with the presence of PrP.Thus, our results on this mouse cell culture model do not seem to support a strong protective role for PrP against transition metal toxicity and also emphasize the necessity of extreme care when comparing cells derived from PrP knock-out and wild type mice.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biochemistry, Biological Research Centre, Hungarian Academy of Sciences, Szeged, Hungary.

ABSTRACT
The interactions of transition metals with the prion protein (PrP) are well-documented and characterized, however, there is no consensus on their role in either the physiology of PrP or PrP-related neurodegenerative disorders. PrP has been reported to protect cells from the toxic stimuli of metals. By employing a cell viability assay, we examined the effects of various concentrations of Cu2+, Zn2+, Mn2+, and Co2+ on Zpl (Prnp-/-) and ZW (Prnp+/+) hippocampus-derived mouse neuronal cells. Prnp-/- Zpl cells were more sensitive to all four metals than PrP-expressing Zw cells. However, when we introduced PrP or only the empty vector into Zpl cells, we could not discern any protective effect associated with the presence of PrP. This observation was further corroborated when assessing the toxic effect of metals by propidium-iodide staining and fluorescence activated cell sorting analysis. Thus, our results on this mouse cell culture model do not seem to support a strong protective role for PrP against transition metal toxicity and also emphasize the necessity of extreme care when comparing cells derived from PrP knock-out and wild type mice.

No MeSH data available.


Related in: MedlinePlus

Sensitivity to transition metal-induced toxicity of ZW 13–2 and Zpl 2–1 cells.Cells were tested for survival after treatment with transition metals for 24 h, assessing cell viability by alamarBlue assay. The cell lines ZW 13–2 (open circles) and Zpl 2–1 (black circles) were treated with increasing concentrations of either Cu2+-Gly (A) or Zn2+ (B), or Mn2+ (C) or Co2+ (D). Values were compared to those of the untreated controls and are presented as percentage. The data represent the means ± standard deviation (S.D.) of minimum 3 independent experiments performed in 5 replicates. *p<0.05, **p<0.01 and ***p<0.001 indicate significant differences between treated and untreated cells; +p<0.05, ++p<0.01 and +++p<0.001 indicate significant differences between ZW 13–2 and Zpl 2–1 cells.
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pone.0139219.g002: Sensitivity to transition metal-induced toxicity of ZW 13–2 and Zpl 2–1 cells.Cells were tested for survival after treatment with transition metals for 24 h, assessing cell viability by alamarBlue assay. The cell lines ZW 13–2 (open circles) and Zpl 2–1 (black circles) were treated with increasing concentrations of either Cu2+-Gly (A) or Zn2+ (B), or Mn2+ (C) or Co2+ (D). Values were compared to those of the untreated controls and are presented as percentage. The data represent the means ± standard deviation (S.D.) of minimum 3 independent experiments performed in 5 replicates. *p<0.05, **p<0.01 and ***p<0.001 indicate significant differences between treated and untreated cells; +p<0.05, ++p<0.01 and +++p<0.001 indicate significant differences between ZW 13–2 and Zpl 2–1 cells.

Mentions: To test whether there is any protective role of PrPC against transition metals-induced toxicity in general we aimed to assess if the two cell lines differed in susceptibility to Cu2+, Zn2+, Mn2+ or Co2+ treatments, using a range of concentrations of each metal ion and an alamarBlue-based cell viability assay. The concentracion rage where the cells proved to be sensitive to metal ion after a 24 h ttreatment was above 200 μM for copper, manganese and cobalt, and above 50 μM in the case of zinc (Fig 2). Such concentration ranges of metals at which the toxicity is observable was also found in the case of other cell lines [57,58,69]. Among the four metal ions, cells proved to be least responsive to Cu2+ treatment, during which cell viability started to decrease significantly only at 500 μM dose compared to untreated cells in case of both cell lines (Fig 2A). Both cell lines are most sensitive to Zn2+ treatment showing significant differences in the number of surviving cells compared to the untreated controls at as little as 100 μM dose (Fig 2B). Nevertheless, Zpl 2–1 cells were significantly more susceptible to Cu2+, Zn2+, Mn2+ and Co2+ toxicities than ZW 13–2 cells at all concentrations of the metal ions that fall into the toxic ranges for the cells (Fig 2).


Prion Protein Does Not Confer Resistance to Hippocampus-Derived Zpl Cells against the Toxic Effects of Cu2+, Mn2+, Zn2+ and Co2+ Not Supporting a General Protective Role for PrP in Transition Metal Induced Toxicity.

Cingaram PK, Nyeste A, Dondapati DT, Fodor E, Welker E - PLoS ONE (2015)

Sensitivity to transition metal-induced toxicity of ZW 13–2 and Zpl 2–1 cells.Cells were tested for survival after treatment with transition metals for 24 h, assessing cell viability by alamarBlue assay. The cell lines ZW 13–2 (open circles) and Zpl 2–1 (black circles) were treated with increasing concentrations of either Cu2+-Gly (A) or Zn2+ (B), or Mn2+ (C) or Co2+ (D). Values were compared to those of the untreated controls and are presented as percentage. The data represent the means ± standard deviation (S.D.) of minimum 3 independent experiments performed in 5 replicates. *p<0.05, **p<0.01 and ***p<0.001 indicate significant differences between treated and untreated cells; +p<0.05, ++p<0.01 and +++p<0.001 indicate significant differences between ZW 13–2 and Zpl 2–1 cells.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591282&req=5

pone.0139219.g002: Sensitivity to transition metal-induced toxicity of ZW 13–2 and Zpl 2–1 cells.Cells were tested for survival after treatment with transition metals for 24 h, assessing cell viability by alamarBlue assay. The cell lines ZW 13–2 (open circles) and Zpl 2–1 (black circles) were treated with increasing concentrations of either Cu2+-Gly (A) or Zn2+ (B), or Mn2+ (C) or Co2+ (D). Values were compared to those of the untreated controls and are presented as percentage. The data represent the means ± standard deviation (S.D.) of minimum 3 independent experiments performed in 5 replicates. *p<0.05, **p<0.01 and ***p<0.001 indicate significant differences between treated and untreated cells; +p<0.05, ++p<0.01 and +++p<0.001 indicate significant differences between ZW 13–2 and Zpl 2–1 cells.
Mentions: To test whether there is any protective role of PrPC against transition metals-induced toxicity in general we aimed to assess if the two cell lines differed in susceptibility to Cu2+, Zn2+, Mn2+ or Co2+ treatments, using a range of concentrations of each metal ion and an alamarBlue-based cell viability assay. The concentracion rage where the cells proved to be sensitive to metal ion after a 24 h ttreatment was above 200 μM for copper, manganese and cobalt, and above 50 μM in the case of zinc (Fig 2). Such concentration ranges of metals at which the toxicity is observable was also found in the case of other cell lines [57,58,69]. Among the four metal ions, cells proved to be least responsive to Cu2+ treatment, during which cell viability started to decrease significantly only at 500 μM dose compared to untreated cells in case of both cell lines (Fig 2A). Both cell lines are most sensitive to Zn2+ treatment showing significant differences in the number of surviving cells compared to the untreated controls at as little as 100 μM dose (Fig 2B). Nevertheless, Zpl 2–1 cells were significantly more susceptible to Cu2+, Zn2+, Mn2+ and Co2+ toxicities than ZW 13–2 cells at all concentrations of the metal ions that fall into the toxic ranges for the cells (Fig 2).

Bottom Line: By employing a cell viability assay, we examined the effects of various concentrations of Cu2+, Zn2+, Mn2+, and Co2+ on Zpl (Prnp-/-) and ZW (Prnp+/+) hippocampus-derived mouse neuronal cells.However, when we introduced PrP or only the empty vector into Zpl cells, we could not discern any protective effect associated with the presence of PrP.Thus, our results on this mouse cell culture model do not seem to support a strong protective role for PrP against transition metal toxicity and also emphasize the necessity of extreme care when comparing cells derived from PrP knock-out and wild type mice.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biochemistry, Biological Research Centre, Hungarian Academy of Sciences, Szeged, Hungary.

ABSTRACT
The interactions of transition metals with the prion protein (PrP) are well-documented and characterized, however, there is no consensus on their role in either the physiology of PrP or PrP-related neurodegenerative disorders. PrP has been reported to protect cells from the toxic stimuli of metals. By employing a cell viability assay, we examined the effects of various concentrations of Cu2+, Zn2+, Mn2+, and Co2+ on Zpl (Prnp-/-) and ZW (Prnp+/+) hippocampus-derived mouse neuronal cells. Prnp-/- Zpl cells were more sensitive to all four metals than PrP-expressing Zw cells. However, when we introduced PrP or only the empty vector into Zpl cells, we could not discern any protective effect associated with the presence of PrP. This observation was further corroborated when assessing the toxic effect of metals by propidium-iodide staining and fluorescence activated cell sorting analysis. Thus, our results on this mouse cell culture model do not seem to support a strong protective role for PrP against transition metal toxicity and also emphasize the necessity of extreme care when comparing cells derived from PrP knock-out and wild type mice.

No MeSH data available.


Related in: MedlinePlus