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Characteristics of the association of nerve growth factor with primary cultures of rat sympathetic neurons

โœ Scribed by Edward Hawrot


Publisher
John Wiley and Sons
Year
1982
Tongue
English
Weight
660 KB
Volume
8
Category
Article
ISSN
0360-4012

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โœฆ Synopsis


Abstract

Longโ€term primary cultures of rat sympathetic neurons require NGF for survival and development. The kinetics of the interaction of ^125^Iโ€NGF with sympathetic neuron cultures suggests the presence of diffusional barriers preventing a determination of true dissociation and association rate constants. Although the observed rate constants do not accurately reflect the microscopic interaction of NGF with receptor, the ratio of the observed rate constants does provide a good estimate of the K~D~. This value (1 ร— 10^โˆ’9^M) agrees with earlier steady state measurements of the K~D~. The association of ^125^Iโ€NGF with neuronal cultures is temperatureโ€dependent with internalization and retrograde transport occurring at 37ยฐC. The retrograde transport of ^125^Iโ€NGF in compartmentalized neuronal cultures is concentration dependent and saturates at about 100 ng/ml (4 ร— 10^โˆ’9^M). The amount of ^125^Iโ€NGF accumulated by retrograde transport appears to be increased subsequent to a period of NGFโ€starvation. The increase in uptake does not appear to be due to an increase in NGF receptor number since the number of binding sites is not greatly increased upon NGF starvation.


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