Bovine Chromaffin Cells: Culture and Fluorescence Assay for Secretion
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Over the last four decades, chromaffin cells originating from the adrenal medulla have been probably one of the most popular cell models to study neurosecretion at the molecular level. Accordingly, numerous seminal discoveries in the field, including the characterization of role of the cytoskeleton, fusogenic lipids, and soluble N-ethylmaleimide-sensitivefactor attachment protein receptor (SNARE) proteins, have been made using this model. In this chapter, we describe a standard method currently used to isolate and culture bovine chromaffin cells, and we illustrate a catecholamine secretion assay based on the successive transformation of adrenaline into adrenochrome and adrenolutine for fluorescence measurements. We also provide some guidelines for efficient cell recovery and for the use of this assay in the laboratory.
Key wordsChromaffin cells Fluorescence assay Catecholamine secretion
We thank Dr. F.A. Meunier (University of Queensland, Brisbane, Australia) and Dr. Robert D. Burgoyne (University of Liverpool) for sharing an original fluorescence assay for catecholamine secretion. We are indebted to the Abattoir Municipal of Haguenau (France) for providing us with the fresh bovine adrenal glands. This work was supported by Fondation pour la Recherche Médicale and the ANR grant (ANR-19-CE44-0019) to NV.
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