In Vitro and Ex Vivo Evaluation of Smart Infra-Red Fluorescent Caspase-3 Probes for Molecular Imaging of Cardiovascular Apoptosis

Manuelle Debunne 1 Christophe Portal 2 Bruno Delest 2 Ebba Brakenhielm 1 Françoise Lallemand 1 Jean-Paul Henry 1 Heidi Ligeret 2 Pauline Noack 2 Marc Massonneau 2 Anthony Romieu 3 Pierre-Yves Renard 3 Christian Thuillez 1 Vincent Richard 1, *
* Corresponding author
1 Pharmacologie des Dysfonctionnements Endotheliaux et Myocardiques
2 QUIDD / Technopôle du Madrillet - QUantitative Imaging in Drug Development [Saint-Etienne du Rouvray]
3 COBRA - Chimie Organique et Bioorganique : Réactivité et Analyse
Abstract : PURPOSE: The aim of this paper is to develop new optical bioprobes for the imaging of apoptosis. PROCEDURE: We developed quenched near-infrared probes which become fluorescent upon cleavage by caspase-3, the key regulatory enzyme of apoptosis. RESULTS: Probes were shown to be selectively cleaved by recombinant caspase-3. Apoptosis of cultured endothelial cells was associated with an increased fluorescent signal for the cleaved probes, which colocalized with caspase-3 and was reduced by the addition of a caspase-3 inhibitor. Flow cytometry demonstrated a similar profile between the cleaved probes and annexin V. Ex vivo experiments showed that sections of hearts obtained from mice treated with the proapoptotic drug doxorubicin displayed an increase in the fluorescent signal for the cleaved probes, which was reduced by a caspase-3 inhibitor. CONCLUSION: We demonstrated the capacity of these novel probes to detect apoptosis by optical imaging in vitro and ex vivo.
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Manuelle Debunne, Christophe Portal, Bruno Delest, Ebba Brakenhielm, Françoise Lallemand, et al.. In Vitro and Ex Vivo Evaluation of Smart Infra-Red Fluorescent Caspase-3 Probes for Molecular Imaging of Cardiovascular Apoptosis. International Journal of Molecular Imaging, Hindawi Publishing Corporation, 2011, 2011, pp.413290. ⟨10.1155/2011/413290⟩. ⟨inserm-02296580⟩

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