Glutamatergic Excitation and GABA Release from a Transplantable Cell Line

Cell Transplantation

Article

Jose Abraham Márquez-Ramos¹  Fernando Peña¹  Mauricio Díaz-Muñoz²  Manuel B. Aguilar²  Magda Giordano³  Jana Mejía-Toiber³ 

[1] Departamento de Farmacobiología, CINVESTAV-Sur. Calzada de los Tenorios 235, Delegación Tlalpan, México;Departamento de Neurobiología Celular y Molecular, Instituto de Neurobiología, Universidad Nacional Autónoma de México, Quéretaro, México;Departamento de Neurobiología Conductual y Cognitiva, Instituto de Neurobiología, Universidad Nacional Autónoma de México, Querétaro, México;

关键词: Glutamate decarboxylase (GAD);    Cell therapy;    GABA uptake;    Exocytosis;    Calcium transients;   

DOI  :  10.3727/096368910X509059

 received in 2009-09-23, accepted in 2010-05-11,  发布年份 2010

来源: Sage Journals

【 摘 要 】

The cell line M213-2O CL-4 was derived from cell line M213-2O and further modified to express human glutamate decarboxylase (hGAD-67), the enzyme that synthesizes GABA. Brain transplants of this cell line in animal models of epilepsy have been shown to modulate seizures. However, the mechanisms that underlie such actions are unknown. The purpose of the present study was to characterize this cell line and its responsiveness to several depolarizing conditions, in order to better understand how these cells exert their effects. Intracellular GABA levels were 34-fold higher and GAD activity was 16-fold higher in clone M213-2O CL-4 than in M213-2O. Both cell lines could take up [3H]GABA in vitro, and this uptake was prevented by nipecotic acid. By combining GABA release measurements and calcium imaging in vitro, we found that high extracellular K+, zero Mg2+, or glutamate activated M213-2O CL-4 cells and resulted in GABA release. The response to glutamate appeared to be mediated by AMPA/NMDA-like receptors. High KCl-induced GABA release was prevented when a Ca2+-free Krebs solution was used, suggesting an exocytotic-like mechanism. These results indicate that the cell line M213-2O CL-4 synthesizes, releases, and takes up GABA in vitro, and can be activated by depolarizing stimuli.

【 授权许可】

Unknown   
© 2010 Cognizant Comm. Corp.

【 图 表 】

Figure 1

Figure 3.

Figure 2.

Figure 5.

Figure 3.

Figure 1.

Figure 4.