Mechanism of Uptake and Retrograde Axonal Transport of Noradrenaline in Sympathetic Neurons.pdfVIP

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Mechanism of Uptake and Retrograde Axonal Transport of Noradrenaline in Sympathetic Neurons.pdf

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Mechanism of Uptake and Retrograde Axonal Transport of Noradrenaline in Sympathetic Neurons

Mechanism of Uptake and Retrograde Axonal Transport of Noradrenaline in Sympathetic Neurons in Culture: Reserpine- resistant Large Dense-core Vesicles as Transport Vehicles MARTIN E. SCHWAB and HANS THOENEN, with the technical assistance of CHRISTIANE MULLER. Department of Neurochemistry, Max-Planck-lnstitute for Psychiatry, D-8033 Martinsried, Federal Republic of Germany ABSTRACT The uptake and retrograde transport of noradrenaline (NA) within the axons of sympathetic neurons was investigated in an in vitro system. Dissociated neurons from the sympathetic ganglia of newborn rats were cultured for 3-6 wk in the absence of non-neuronal cells in a culture dish divided into three chambers. These allowed separate access to the axonal networks and to their cell bodies of origin. [3H]NA (0.5 X 10 -e M), added to the axon chambers, was taken up by the desmethylimipramine- and cocaine-sensitive neuronal amine uptake mechanisms, and a substantial part was rapidly transported retrogradely along the axons to the nerve cell bodies. This transport was blocked by vinblastine or colchicine. In contrast with the storage of [3H]NA in the axonal varicosities, which was totally prevented by reserpine (a drug that selectively inactivates the uptake of NA into adrenergic storage vesicles), the retrograde transport of [3H]NA was only slightly diminished by reserpine pretreatment. Electron micro- scopic localization of the NA analogue 5-hydroxydopamine (5-OHDA) indicated that mainly large dense-core vesicles (700-1,200-A diam) are the transport compartment involved. Whereas the majority of small and large vesicles lost their amine dense-core and were unable to take up 5-OHDA after reserpine pretreatment, the population of large dense-core vesicles and some small dense-core vesicles involved in the retrograde transport were resistant to this drug. It, therefore, seems that these vesicles maintained the amine uptake and storage mechanisms characteristic for adrenerg