Existence of an Endogenous Glutamate and Aspartate Transporter in Chinese Hamster Ovary Cells

Abstract

Chinese hamster ovary cells show endogenous high-affinity Na(+)-dependent glutamate transport activity. This transport activity is kinetically similar to a glutamate transporter family strategically expressed in the central nervous system and is pharmacologically unlike glutamate transporter-1 or excitatory amino acid carrier 1. The cDNA of a glutamate/aspartate transporter (GLAST)-like transporter was obtained and analyzed. The deduced amino acid sequence showed high similarity to human, mouse, and rat GLAST. We concluded that a GLAST-like glutamate transporter exists in Chinese hamster ovary cells that might confer the endogenous high-affinity Na(+)-dependent glutamate transport activity evident in these cells.

Expression of Cbl-interacting protein of 85 kDa in MPTP mouse model of Parkinson's disease and 1-methyl-4-phenyl-pyridinium ion-treated dopaminergic SH-SY5Y cells

“…Total RNA was extracted from SH-SY5Y cells using Trizol reagent (Invitrogen, Carlsbad, USA). Reverse transcription was carried out using random primer and Moloney murine leukemia virus reverse transcriptase (Promega, Madison, USA), and real-time PCR was carried out for quantification of CIN85 mRNA on Rotor-Gene 3000 (Corbett Research, Sydney, Australia) based on published methods [18,19]. For plotting a standard curve, serially diluted β-actin and CIN85 cDNA fragments were used in each experiment.…”