ParkinsonCanadaV1, Main, Exploration, bibRecord, 003C55

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 : 003C55 ( Main/Exploration ); précédent : 003C54; suivant : 003C56

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

Auteurs : N. A. Tatton [Canada] ; S. J. Kish [Canada]

Source :

Neuroscience [ 0306-4522 ] ; 1997.

RBID : Pascal:97-0227389

Descripteurs français

Pascal (Inist)
- Locus niger, Apoptose, Parkinson maladie, Neurotoxine, Pathologie expérimentale, Animal, Souris, MPTP.

English descriptors

KwdEn :
- 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine, Acridine Orange, Animal, Animals, Apoptosis, Apoptosis (physiology), Cats, Cell Nucleus (chemistry), Cell Nucleus (pathology), DNA (analysis), Disease Models, Animal, Dopamine Agents, Experimental disease, Fluorescent Dyes, Locus niger, Male, Mice, Mice, Inbred C57BL, Microscopy, Confocal, Mouse, Necrosis, Neurotoxin, Nucleotidyltransferases, Parkinson Disease, Secondary (chemically induced), Parkinson Disease, Secondary (pathology), Parkinson disease, 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

Absfract-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 identify cells with double-stranded DNA breaks. Apoptotic cell death was found to be initiated within 72h 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.

Affiliations:

Canada

Le document en format XML

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<front><div type="abstract" xml:lang="en">Absfract-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 identify cells with double-stranded DNA breaks. Apoptotic cell death was found to be initiated within 72h 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>
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La maladie de Parkinson au Canada (serveur d'exploration)

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

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

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri

	La maladie de Parkinson au Canada (serveur d'exploration)
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