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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

The effect of the presence of PrPC on the susceptibility of cells to transition metal-induced toxicity.Cells were tested for survival after treatment with transition metals for 24 h and cell viability was determined using alamarBlue assay. Values are compared to those of the untreated controls and are expressed as percentage. The cells Zpl 2-1-PrP (open triangles) and Zpl 2-1-vector (black triangles) are compared (panels A through D) during treatments with increasing concentrations of metals, as follows: Cu2+-Gly (A), of Zn2+ (B), Mn2+ (C) and Co2+ (D). The cell lines ZW 13–2 (open circles), Zpl 2–1 (black circles) were treated similarly with increasing concentrations of metals and all four types of cells are compared on panels E through H, as follows: increasing concentration of Cu2+-Gly (E), of Zn2+ (F), Mn2+ (G) and of Co2+(H). The data are presented as 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 on panels A through D; μp<0.05, μμp<0.01 and μμμp<0.001 indicate significant differences between treated Zpl 2-1-PrP cells and treated Zpl 2-1-vector cells; +p<0.05, ++p<0.01 and +++p<0.001 indicate significant differences between the treated ZW 13–2 cells and treated Zpl 2–1 cells; δp<0.05, δδp<0.01 and δδδp<0.001indicate significant differences between the treated ZW 13–2 cells and treated Zpl 2-1-vector cells, and φp<0.05, φφp<0.01 and φφφp<0.001 indicate significant differences between the treated ZW 13–2 cells and treated Zpl 2-1-PrP cells.
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pone.0139219.g004: The effect of the presence of PrPC on the susceptibility of cells to transition metal-induced toxicity.Cells were tested for survival after treatment with transition metals for 24 h and cell viability was determined using alamarBlue assay. Values are compared to those of the untreated controls and are expressed as percentage. The cells Zpl 2-1-PrP (open triangles) and Zpl 2-1-vector (black triangles) are compared (panels A through D) during treatments with increasing concentrations of metals, as follows: Cu2+-Gly (A), of Zn2+ (B), Mn2+ (C) and Co2+ (D). The cell lines ZW 13–2 (open circles), Zpl 2–1 (black circles) were treated similarly with increasing concentrations of metals and all four types of cells are compared on panels E through H, as follows: increasing concentration of Cu2+-Gly (E), of Zn2+ (F), Mn2+ (G) and of Co2+(H). The data are presented as 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 on panels A through D; μp<0.05, μμp<0.01 and μμμp<0.001 indicate significant differences between treated Zpl 2-1-PrP cells and treated Zpl 2-1-vector cells; +p<0.05, ++p<0.01 and +++p<0.001 indicate significant differences between the treated ZW 13–2 cells and treated Zpl 2–1 cells; δp<0.05, δδp<0.01 and δδδp<0.001indicate significant differences between the treated ZW 13–2 cells and treated Zpl 2-1-vector cells, and φp<0.05, φφp<0.01 and φφφp<0.001 indicate significant differences between the treated ZW 13–2 cells and treated Zpl 2-1-PrP cells.

Mentions: To test whether the sole presence of PrP is responsible for the different susceptibilities of PrP-expressing ZW 13–2 compared to Prnp−/−Zpl 2–1 against metal toxicity, we treated Zpl 2-1-PrP and Zpl 2-1-vector cells characterized above (Fig 3) with Cu2+, Zn2+, Mn2+ and Co2+ at the same concentrations as on Fig 2 and assessed their survival using alamarBlue-based assay (Fig 4A, 4B, 4C and 4D). Treatment with Cu2+, Zn2+, Mn2+ and Co2+ decreased significantly the cell viabilities compared to the untreated controls and at the same metal ion concentrations as in the case of ZW 13–2 and Zpl 2–1 cells. Interestingly, Zpl 2-1-PrP and Zpl 2-1-vector cells exhibit similar sensitivities, without any significant difference, in case of each metal concentration tested. These findings argue that PrP is not the sole reason for the increased resistance of ZW 13–2 cells compared to Zpl 2–1 cells. Further, when the cell survival data of all four types of cells, ZW 13–2, Zpl 2–1, Zpl 2-1-PrP and Zpl 2-1-vector are compared (Fig 4E, 4F, 4G and 4H) it can be seen that in case of Cu+2, Zn+2, and Co+2 the viabilities of Zpl 2-1-PrP and Zpl 2-1-vector cells are significantly lower than that of ZW 13–2 cells, whereas, no significant difference is observed in case of Mn+2 treatment.


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)

The effect of the presence of PrPC on the susceptibility of cells to transition metal-induced toxicity.Cells were tested for survival after treatment with transition metals for 24 h and cell viability was determined using alamarBlue assay. Values are compared to those of the untreated controls and are expressed as percentage. The cells Zpl 2-1-PrP (open triangles) and Zpl 2-1-vector (black triangles) are compared (panels A through D) during treatments with increasing concentrations of metals, as follows: Cu2+-Gly (A), of Zn2+ (B), Mn2+ (C) and Co2+ (D). The cell lines ZW 13–2 (open circles), Zpl 2–1 (black circles) were treated similarly with increasing concentrations of metals and all four types of cells are compared on panels E through H, as follows: increasing concentration of Cu2+-Gly (E), of Zn2+ (F), Mn2+ (G) and of Co2+(H). The data are presented as 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 on panels A through D; μp<0.05, μμp<0.01 and μμμp<0.001 indicate significant differences between treated Zpl 2-1-PrP cells and treated Zpl 2-1-vector cells; +p<0.05, ++p<0.01 and +++p<0.001 indicate significant differences between the treated ZW 13–2 cells and treated Zpl 2–1 cells; δp<0.05, δδp<0.01 and δδδp<0.001indicate significant differences between the treated ZW 13–2 cells and treated Zpl 2-1-vector cells, and φp<0.05, φφp<0.01 and φφφp<0.001 indicate significant differences between the treated ZW 13–2 cells and treated Zpl 2-1-PrP cells.
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Related In: Results  -  Collection

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Show All Figures
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pone.0139219.g004: The effect of the presence of PrPC on the susceptibility of cells to transition metal-induced toxicity.Cells were tested for survival after treatment with transition metals for 24 h and cell viability was determined using alamarBlue assay. Values are compared to those of the untreated controls and are expressed as percentage. The cells Zpl 2-1-PrP (open triangles) and Zpl 2-1-vector (black triangles) are compared (panels A through D) during treatments with increasing concentrations of metals, as follows: Cu2+-Gly (A), of Zn2+ (B), Mn2+ (C) and Co2+ (D). The cell lines ZW 13–2 (open circles), Zpl 2–1 (black circles) were treated similarly with increasing concentrations of metals and all four types of cells are compared on panels E through H, as follows: increasing concentration of Cu2+-Gly (E), of Zn2+ (F), Mn2+ (G) and of Co2+(H). The data are presented as 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 on panels A through D; μp<0.05, μμp<0.01 and μμμp<0.001 indicate significant differences between treated Zpl 2-1-PrP cells and treated Zpl 2-1-vector cells; +p<0.05, ++p<0.01 and +++p<0.001 indicate significant differences between the treated ZW 13–2 cells and treated Zpl 2–1 cells; δp<0.05, δδp<0.01 and δδδp<0.001indicate significant differences between the treated ZW 13–2 cells and treated Zpl 2-1-vector cells, and φp<0.05, φφp<0.01 and φφφp<0.001 indicate significant differences between the treated ZW 13–2 cells and treated Zpl 2-1-PrP cells.
Mentions: To test whether the sole presence of PrP is responsible for the different susceptibilities of PrP-expressing ZW 13–2 compared to Prnp−/−Zpl 2–1 against metal toxicity, we treated Zpl 2-1-PrP and Zpl 2-1-vector cells characterized above (Fig 3) with Cu2+, Zn2+, Mn2+ and Co2+ at the same concentrations as on Fig 2 and assessed their survival using alamarBlue-based assay (Fig 4A, 4B, 4C and 4D). Treatment with Cu2+, Zn2+, Mn2+ and Co2+ decreased significantly the cell viabilities compared to the untreated controls and at the same metal ion concentrations as in the case of ZW 13–2 and Zpl 2–1 cells. Interestingly, Zpl 2-1-PrP and Zpl 2-1-vector cells exhibit similar sensitivities, without any significant difference, in case of each metal concentration tested. These findings argue that PrP is not the sole reason for the increased resistance of ZW 13–2 cells compared to Zpl 2–1 cells. Further, when the cell survival data of all four types of cells, ZW 13–2, Zpl 2–1, Zpl 2-1-PrP and Zpl 2-1-vector are compared (Fig 4E, 4F, 4G and 4H) it can be seen that in case of Cu+2, Zn+2, and Co+2 the viabilities of Zpl 2-1-PrP and Zpl 2-1-vector cells are significantly lower than that of ZW 13–2 cells, whereas, no significant difference is observed in case of Mn+2 treatment.

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