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Two-Photon Analysis of Lead Accumulation in Rat Cerebellar Granule Neurons

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Lead (Pb2+) is a common pollutant and potent central neurotoxin. We have studied its pathways of permeation by two-photon fluorescence microscopy in rat cerebellar granule neurons loaded with the fluorescent dye indo-1. Pb2+ binds indo-1 with high affinity acting as a quencher. Its permeation through the neuronal membrane was indicated by a decrease of the fluorescence emission, which occurred even in resting condition. In the presence of 20 μM Pb2+, uptake reached a plateau level (≈45% of initial fluorescence) in 4 min and was partially antagonized by 25 μM lanthanum. Subsequent addition of a membrane permeant ionophore caused a further (>70%) quenching of the dye, suggesting that previous saturation was due to inactivation of the transport system. Intracellular Pb2+ concentrations were evaluated from the fluorescence intensity and this estimate indicated that the concentration of free Pb2+ sufficient to inactivate the transport system is close to 50 pM.

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Authors and Affiliations

  1. INFM, Dipartimento di Fisica, Università di Genova, via Dodecaneso, 33, 16146, Genova, Italy

    Alessandro Esposito, Mauro Robello & Francesca Pellistri

  2. Istituto di Biofisica, CNR, via De Marini 6, 16149, Genova, Italy

    Carla Marchetti

  3. Cell Biophysics Group, European Neuroscience Institute, Göttingen Waldweg 33, D-37073, Göttingen, Germany

    Alessandro Esposito

Authors
  1. Alessandro Esposito
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  2. Mauro Robello
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  3. Francesca Pellistri
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  4. Carla Marchetti
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Correspondence to Mauro Robello.

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Esposito, A., Robello, M., Pellistri, F. et al. Two-Photon Analysis of Lead Accumulation in Rat Cerebellar Granule Neurons . Neurochem Res 30, 949–954 (2005). https://doi.org/10.1007/s11064-005-5980-y

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