A method for objectively quantifying propidium iodide exclusion in organotypic hippocampal slice cultures.
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| Title: | A method for objectively quantifying propidium iodide exclusion in organotypic hippocampal slice cultures. |
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| Authors: | Happ, Denise F.1, Tasker, R. Andrew1,2 tasker@upei.ca |
| Source: | Journal of Neuroscience Methods. Aug2016, Vol. 269, p1-5. 5p. |
| Subjects: | Propidium iodide, Hippocampus (Brain), Neurons, Methyl aspartate, Cell death, Cell survival, Wounds & injuries |
| Abstract: | Background Organotypic hippocampal slice cultures (OHSCs) are an attractive in vitro model to examine mechanisms of neuronal injury, because the normal hippocampal architecture, function and cellular diversity are mostly preserved. The effects of exposure to excitotoxins such as N -methyl- d -aspartate (NMDA) on cell viability can be determined by propidium iodide (PI) staining. New method We describe a simple method to objectively quantify cell death in NMDA exposed slice cultures using PI that provides a standardized means of quantifying cell death in hippocampal subfields without the need to induce maximal cell death in each slice. The method employs separation of subfields using simple landmarks and densitometric quantification of PI intensity in 10 template-oriented counting fields. Results We show that exposure to increasing concentrations of NMDA results in a dose-dependent increase in PI uptake. Additionally, our method facilitates the comparison of cell death in different hippocampal subfields, such as dentate gyrus, CA1 and CA3. Our results show marked differences of PI uptake in the hippocampal regions with the CA1 area being most sensitive to NMDA-induced injury. Comparison with existing method(s) The method provides a standardized format for quantifying PI exclusion in OHSCs that can be applied to cultures of differing shapes and sizes, permits comparisons between hippocampal subfields and does not require induction of maximal cell death. Conclusion The method of quantifying PI uptake described herein allows for an objective, quantitative and reproducible analysis and comparison of cell death in distinct regions of OHSCs. [ABSTRACT FROM AUTHOR] |
| Copyright of Journal of Neuroscience Methods is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) | |
| Database: | Engineering Source |
| FullText | Text: Availability: 0 |
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| Header | DbId: egs DbLabel: Engineering Source An: 116404875 AccessLevel: 6 PubType: Academic Journal PubTypeId: academicJournal PreciseRelevancyScore: 0 |
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| Items | – Name: Title Label: Title Group: Ti Data: A method for objectively quantifying propidium iodide exclusion in organotypic hippocampal slice cultures. – Name: Author Label: Authors Group: Au Data: <searchLink fieldCode="AR" term="%22Happ%2C+Denise+F%2E%22">Happ, Denise F.</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Tasker%2C+R%2E+Andrew%22">Tasker, R. Andrew</searchLink><relatesTo>1,2</relatesTo><i> tasker@upei.ca</i> – Name: TitleSource Label: Source Group: Src Data: <searchLink fieldCode="JN" term="%22Journal+of+Neuroscience+Methods%22">Journal of Neuroscience Methods</searchLink>. Aug2016, Vol. 269, p1-5. 5p. – Name: Subject Label: Subjects Group: Su Data: <searchLink fieldCode="DE" term="%22Propidium+iodide%22">Propidium iodide</searchLink><br /><searchLink fieldCode="DE" term="%22Hippocampus+%28Brain%29%22">Hippocampus (Brain)</searchLink><br /><searchLink fieldCode="DE" term="%22Neurons%22">Neurons</searchLink><br /><searchLink fieldCode="DE" term="%22Methyl+aspartate%22">Methyl aspartate</searchLink><br /><searchLink fieldCode="DE" term="%22Cell+death%22">Cell death</searchLink><br /><searchLink fieldCode="DE" term="%22Cell+survival%22">Cell survival</searchLink><br /><searchLink fieldCode="DE" term="%22Wounds+%26+injuries%22">Wounds & injuries</searchLink> – Name: Abstract Label: Abstract Group: Ab Data: Background Organotypic hippocampal slice cultures (OHSCs) are an attractive in vitro model to examine mechanisms of neuronal injury, because the normal hippocampal architecture, function and cellular diversity are mostly preserved. The effects of exposure to excitotoxins such as N -methyl- d -aspartate (NMDA) on cell viability can be determined by propidium iodide (PI) staining. New method We describe a simple method to objectively quantify cell death in NMDA exposed slice cultures using PI that provides a standardized means of quantifying cell death in hippocampal subfields without the need to induce maximal cell death in each slice. The method employs separation of subfields using simple landmarks and densitometric quantification of PI intensity in 10 template-oriented counting fields. Results We show that exposure to increasing concentrations of NMDA results in a dose-dependent increase in PI uptake. Additionally, our method facilitates the comparison of cell death in different hippocampal subfields, such as dentate gyrus, CA1 and CA3. Our results show marked differences of PI uptake in the hippocampal regions with the CA1 area being most sensitive to NMDA-induced injury. Comparison with existing method(s) The method provides a standardized format for quantifying PI exclusion in OHSCs that can be applied to cultures of differing shapes and sizes, permits comparisons between hippocampal subfields and does not require induction of maximal cell death. Conclusion The method of quantifying PI uptake described herein allows for an objective, quantitative and reproducible analysis and comparison of cell death in distinct regions of OHSCs. [ABSTRACT FROM AUTHOR] – Name: AbstractSuppliedCopyright Label: Group: Ab Data: <i>Copyright of Journal of Neuroscience Methods is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.) |
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| RecordInfo | BibRecord: BibEntity: Identifiers: – Type: doi Value: 10.1016/j.jneumeth.2016.05.006 Languages: – Code: eng Text: English PhysicalDescription: Pagination: PageCount: 5 StartPage: 1 Subjects: – SubjectFull: Propidium iodide Type: general – SubjectFull: Hippocampus (Brain) Type: general – SubjectFull: Neurons Type: general – SubjectFull: Methyl aspartate Type: general – SubjectFull: Cell death Type: general – SubjectFull: Cell survival Type: general – SubjectFull: Wounds & injuries Type: general Titles: – TitleFull: A method for objectively quantifying propidium iodide exclusion in organotypic hippocampal slice cultures. Type: main BibRelationships: HasContributorRelationships: – PersonEntity: Name: NameFull: Happ, Denise F. – PersonEntity: Name: NameFull: Tasker, R. Andrew IsPartOfRelationships: – BibEntity: Dates: – D: 30 M: 08 Text: Aug2016 Type: published Y: 2016 Identifiers: – Type: issn-print Value: 01650270 Numbering: – Type: volume Value: 269 Titles: – TitleFull: Journal of Neuroscience Methods Type: main |
| ResultId | 1 |
