In situ detection of apoptotic nuclei in the substantia nigra compacta of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated mice using terminal deoxynucleotidyl transferase labelling and acridine orange staining.
Identifieur interne : 002582 ( Ncbi/Checkpoint ); précédent : 002581; suivant : 002583In situ detection of apoptotic nuclei in the substantia nigra compacta of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated mice using terminal deoxynucleotidyl transferase labelling and acridine orange staining.
Auteurs : N A Tatton [Canada] ; S J KishSource :
- Neuroscience [ 0306-4522 ] ; 1997.
English descriptors
- KwdEn :
- 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine, Acridine Orange, Animals, Apoptosis (physiology), Cats, Cell Nucleus (chemistry), Cell Nucleus (pathology), DNA (analysis), Disease Models, Animal, Dopamine Agents, Fluorescent Dyes, Male, Mice, Mice, Inbred C57BL, Microscopy, Confocal, Necrosis, Nucleotidyltransferases, Parkinson Disease, Secondary (chemically induced), Parkinson Disease, Secondary (pathology), Staining and Labeling (methods), Substantia Nigra (cytology).
- MESH :
- chemical , analysis : DNA.
- chemical : 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine, Acridine Orange, Dopamine Agents, Fluorescent Dyes, Nucleotidyltransferases.
- chemically induced : Parkinson Disease, Secondary.
- chemistry : Cell Nucleus.
- cytology : Substantia Nigra.
- methods : Staining and Labeling.
- pathology : Cell Nucleus, Parkinson Disease, Secondary.
- physiology : Apoptosis.
- Animals, Cats, Disease Models, Animal, Male, Mice, Mice, Inbred C57BL, Microscopy, Confocal, Necrosis.
Abstract
The neurotoxin, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) was used to generate a dose-dependent cell death of dopaminergic nigral neurons in the C57B1 mouse. Mice were injected with a total cumulative dose of 150 mg/kg of MPTP delivered over five days and killed at different time points both during and after the toxin injections. Two independent histological methods were used to determine whether the dopaminergic nigral neurons died via an apoptotic mechanism. In situ end-labelling with terminal deoxynucleotidyl transferase was used on paraformaldehyde-fixed, serial, frozen sections to identity cells with double-stranded DNA breaks. Apoptotic cell death was found to be initiated within 72 h of the first injection of the neurotoxin and peaked 24 h after the final MPTP injection. The metachromatic fluorochrome, Acridine Orange, was used on alternate sections to provide structural confirmation of the nuclear chromatin "clumping" considered to be representative of apoptosis. Confocal laser imaging combined with deconvolution techniques was used to resolve the fluorescent signal emitted by the in situ Acridine Orange-DNA complexes. The number of Acridine Orange-stained nuclei demonstrating chromatin clumping was identical to that of the positive in situ end-labelled nuclei observed over a 25 day period. Based upon these two independent methods of assessing apoptosis in situ, we conclude that a 150 mg/kg dose of MPTP can elicit apoptotic cell death in nigral dopaminergic neurons of the C57B1 mouse.
PubMed: 9130785
Affiliations:
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">In situ detection of apoptotic nuclei in the substantia nigra compacta of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated mice using terminal deoxynucleotidyl transferase labelling and acridine orange staining.</title><author><name sortKey="Tatton, N A" sort="Tatton, N A" uniqKey="Tatton N" first="N A" last="Tatton">N A Tatton</name><affiliation wicri:level="1"><nlm:affiliation>Department of Pharmacology, Dalhousie University, Halifax, Nova Scotia, Canada.</nlm:affiliation><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Pharmacology, Dalhousie University, Halifax, Nova Scotia</wicri:regionArea><wicri:noRegion>Nova Scotia</wicri:noRegion></affiliation></author><author><name sortKey="Kish, S J" sort="Kish, S J" uniqKey="Kish S" first="S J" last="Kish">S J Kish</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="1997">1997</date><idno type="RBID">pubmed:9130785</idno><idno type="pmid">9130785</idno><idno type="wicri:Area/PubMed/Corpus">001831</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001831</idno><idno type="wicri:Area/PubMed/Curation">001831</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">001831</idno><idno type="wicri:Area/PubMed/Checkpoint">001831</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">001831</idno><idno type="wicri:Area/Ncbi/Merge">002582</idno><idno type="wicri:Area/Ncbi/Curation">002582</idno><idno type="wicri:Area/Ncbi/Checkpoint">002582</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">In situ detection of apoptotic nuclei in the substantia nigra compacta of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated mice using terminal deoxynucleotidyl transferase labelling and acridine orange staining.</title><author><name sortKey="Tatton, N A" sort="Tatton, N A" uniqKey="Tatton N" first="N A" last="Tatton">N A Tatton</name><affiliation wicri:level="1"><nlm:affiliation>Department of Pharmacology, Dalhousie University, Halifax, Nova Scotia, Canada.</nlm:affiliation><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Pharmacology, Dalhousie University, Halifax, Nova Scotia</wicri:regionArea><wicri:noRegion>Nova Scotia</wicri:noRegion></affiliation></author><author><name sortKey="Kish, S J" sort="Kish, S J" uniqKey="Kish S" first="S J" last="Kish">S J Kish</name></author></analytic><series><title level="j">Neuroscience</title><idno type="ISSN">0306-4522</idno><imprint><date when="1997" type="published">1997</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine</term><term>Acridine Orange</term><term>Animals</term><term>Apoptosis (physiology)</term><term>Cats</term><term>Cell Nucleus (chemistry)</term><term>Cell Nucleus (pathology)</term><term>DNA (analysis)</term><term>Disease Models, Animal</term><term>Dopamine Agents</term><term>Fluorescent Dyes</term><term>Male</term><term>Mice</term><term>Mice, Inbred C57BL</term><term>Microscopy, Confocal</term><term>Necrosis</term><term>Nucleotidyltransferases</term><term>Parkinson Disease, Secondary (chemically induced)</term><term>Parkinson Disease, Secondary (pathology)</term><term>Staining and Labeling (methods)</term><term>Substantia Nigra (cytology)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>DNA</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine</term><term>Acridine Orange</term><term>Dopamine Agents</term><term>Fluorescent Dyes</term><term>Nucleotidyltransferases</term></keywords><keywords scheme="MESH" qualifier="chemically induced" xml:lang="en"><term>Parkinson Disease, Secondary</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Cell Nucleus</term></keywords><keywords scheme="MESH" qualifier="cytology" xml:lang="en"><term>Substantia Nigra</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Staining and Labeling</term></keywords><keywords scheme="MESH" qualifier="pathology" xml:lang="en"><term>Cell Nucleus</term><term>Parkinson Disease, Secondary</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Apoptosis</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Cats</term><term>Disease Models, Animal</term><term>Male</term><term>Mice</term><term>Mice, Inbred C57BL</term><term>Microscopy, Confocal</term><term>Necrosis</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The neurotoxin, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) was used to generate a dose-dependent cell death of dopaminergic nigral neurons in the C57B1 mouse. Mice were injected with a total cumulative dose of 150 mg/kg of MPTP delivered over five days and killed at different time points both during and after the toxin injections. Two independent histological methods were used to determine whether the dopaminergic nigral neurons died via an apoptotic mechanism. In situ end-labelling with terminal deoxynucleotidyl transferase was used on paraformaldehyde-fixed, serial, frozen sections to identity cells with double-stranded DNA breaks. Apoptotic cell death was found to be initiated within 72 h of the first injection of the neurotoxin and peaked 24 h after the final MPTP injection. The metachromatic fluorochrome, Acridine Orange, was used on alternate sections to provide structural confirmation of the nuclear chromatin "clumping" considered to be representative of apoptosis. Confocal laser imaging combined with deconvolution techniques was used to resolve the fluorescent signal emitted by the in situ Acridine Orange-DNA complexes. The number of Acridine Orange-stained nuclei demonstrating chromatin clumping was identical to that of the positive in situ end-labelled nuclei observed over a 25 day period. Based upon these two independent methods of assessing apoptosis in situ, we conclude that a 150 mg/kg dose of MPTP can elicit apoptotic cell death in nigral dopaminergic neurons of the C57B1 mouse.</div></front></TEI><affiliations><list><country><li>Canada</li></country></list><tree><noCountry><name sortKey="Kish, S J" sort="Kish, S J" uniqKey="Kish S" first="S J" last="Kish">S J Kish</name></noCountry><country name="Canada"><noRegion><name sortKey="Tatton, N A" sort="Tatton, N A" uniqKey="Tatton N" first="N A" last="Tatton">N A Tatton</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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