Evaluation of the tetracycline‐ and ecdysone‐inducible systems for expression of neurotransmitter receptors in mammalian cells

Establishing a stable cell line that expresses a particular protein of interest is often a laborious and time‐consuming experience. With constitutive expression systems, a gradual loss of the highly expressing clones over a given time span and/or a severe counter‐selection due to toxicity of the expressed protein for the host cell line are major drawbacks. In both cases, inducible expression systems offer a valuable alternative. Over the years, many regulated expression systems have been developed and evaluated. In the present study, we compare the efficiency, the advantages and the drawbacks of a tetracycline‐ and an ecdysone‐inducible system for expression of the reporter protein chloramphenicol acetyltransferase and of different G‐protein‐coupled serotonin (5‐HT) receptors. A high level of expression of different 5‐HT receptors was obtained with the tetracycline‐inducible system. In the cell line L929, which stably expresses the tetracycline‐responsive transactivator, a maximum ligand binding of 20 000 and 9500 fmol/mg protein was measured for the h5‐HT1B and h5‐ht1F receptors, respectively. In the HEK293rtTA cell line, levels of 15 700, 3000, and 9100 fmol bound ligand/mg protein were obtained for the h5‐HT1B, h5‐ht1F and h5‐HT4b receptors, respectively. These high expression levels remained stable for several months of continuous culture. Although the ecdysone‐inducible expression system was useful for tightly regulated expression, the levels were far lower than those obtained with the tetracycline system (e.g. 640 fmol bound ligand/mg protein for the h5‐ht1F receptor in HEK293EcR).

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