Flow cytometric studies of nucleoside transport regulation in single chromaffin cells

FEBS Letters

Alvarez, Alberto¹  Brocklebank, Anne M²  Delicado, Esmerilda G³  Miras-Portugal, Mª Teresa³  Sen, Raquel P³  Wiley, James S² 

[1] Servicio de Citometrı́a de Flujo, Universidad Complutense, 28040 Madrid, Spain;Department of Haematology, Austin Hospital, Heidelberg, Melbourne, Vic. 3084, Australia;Departamento de Bioquı́mica, Facultad de Veterinaria, Universidad Complutense, 28040 Madrid, Spain

关键词: Nucleoside transport;    Nitrobenzylthioinosine;    Flow cytometry;    Chromaffin cell;   

DOI  :  10.1016/S0014-5793(98)00047-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The present paper reveals that a fluorescent derivative of nitrobenzylthioinosine, 5-(SAENTA-x8)-fluorescein, is a highly specific inhibitor of the neural NBTI-sensitive nucleoside transporter. 5-(SAENTA-x8)-fluorescein inhibited adenosine transport and [³H]NBTI binding with a K _i of 4 nM in cultured chromaffin cells. Flow cytometry demonstrated that 5-(SAENTA-x8)-fluorescein specifically interacted with the NBTI-sensitive nucleoside transporters with high affinity (K _D=6 nM). Activation of protein kinases A and C with forskolin or nicotinic receptor agonists, respectively, resulted in 50% inhibition of the fluorescence bound to the cells. Flow cytometry will allow studying nucleoside transport in single cells from heterogeneous neural cell populations.

【 授权许可】

Unknown