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Dynamic analysis of apoptosis in primary cortical neurons by fixed- and real-time cytofluorometry

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Abstract

We describe here a cytofluorometric technology for the characterization of decision, execution, and degradation steps of neuronal apoptosis. Multiparametric flow cytometry was developped and combined to detailled fluorescence microscopy observations to establish the chronology and hierarchy of death-related events: neuron morphological changes, mitochondrial transmembrane potential (Δ Ψ m ) collapse, caspase-3 and -9 activation, phosphatidyl-serine exposure, nuclear dismantling and final plasma membrane permeabilization. Moreover, we developped a reliable real-time flow cytometric monitoring of Δ Ψ m and plasma membrane integrity in response to neurotoxic insults including MPTP treatment. Taking advantage of recently developped specific fluorescent probes and a third generation pan-caspase inhibitor, this integrated approach will be pertinent to study the cell biology of neuronal apoptosis and to characterize new neuro-toxic/protective molecules.

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Authors and Affiliations

  1. Theraptosis Research Laboratory, THERAPTOSIS S.A., Pasteur Biotop, Institut Pasteur, 75015, Paris, France

    H. Lecoeur, D. Chauvier, A. Langonné, D. Rebouillat, L. Edelman & E. Jacotot

  2. Neurobiologie des processus adaptatifs, UMR 7102 CNRS, Université Pierre et Marie-Curie, 75005, Paris, France

    B. Brugg & J. Mariani

  3. Technologie Cellulaire anti-tumorale, Institut Pasteur, 75015, Paris, France

    L. Edelman

Authors
  1. H. Lecoeur
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  2. D. Chauvier
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  3. A. Langonné
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  4. D. Rebouillat
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  5. B. Brugg
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  6. J. Mariani
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  7. L. Edelman
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  8. E. Jacotot
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Correspondence to H. Lecoeur.

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Lecoeur, H., Chauvier, D., Langonné, A. et al. Dynamic analysis of apoptosis in primary cortical neurons by fixed- and real-time cytofluorometry. Apoptosis 9, 157–169 (2004). https://doi.org/10.1023/B:APPT.0000018798.03705.69

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  • DOI: https://doi.org/10.1023/B:APPT.0000018798.03705.69

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