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  Low Ca2+ levels in the culture media support the heterologous expression of insect odorant receptor proteins in HEK cells

Miazzi, F., Schulze, H.-C., Zhang, L., Kaltofen, S., Hansson, B. S., & Wicher, D. (2018). Low Ca2+ levels in the culture media support the heterologous expression of insect odorant receptor proteins in HEK cells. Journal of Neuroscience Methods, 312, 122-125. doi:10.1016/j.jneumeth.2018.11.021.

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HAN318.pdf (Publisher version), 743KB
 
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Miazzi, Fabio1, 2, Author              
Schulze, Hannes-Claudius1, Author              
Zhang, Liwei1, Author              
Kaltofen, Sabine1, Author              
Hansson, Bill S.1, Author              
Wicher, Dieter1, Author              
Affiliations:
1Department of Evolutionary Neuroethology, Prof. B. S. Hansson, MPI for Chemical Ecology, Max Planck Society, ou_421894              
2IMPRS on Ecological Interactions, MPI for Chemical Ecology, Max Planck Society, Jena, DE, ou_421900              

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 Abstract: Background: Heterologous expression of insect odorant receptors (ORs) in mammalian or insect cells is challenging due to the insufficient intracellular trafficking of ORs and their ability to form leak ion channels. New Method: We tested whether reducing the Ca2+ levels in the cell culture medium after electroporation by means of a Dulbecco's Modified Eagle Medium (DMEM) without calcium, in a 1:1 ratio with Ham's F12 nutrient mixture, together with 10% fetal calf serum, can improve the success rate of insect OR expression in HEK293 cells. Results: We show that a reduced extracellular Ca2+ level supports functional expression of insect ORs by increasing the fraction of cells responding to the co-receptor agonist VUAA1 and by reducing the intracellular Ca2+ base level of transfected cells. Comparison with existing method(s): A DMEM formula without calcium outperforms standard DMEM in a 1:1 ratio with Ham's F12 mix and 10% serum, when culturing HEK293 cells transiently expressing insect OR proteins. Conclusions: Reducing the extracellular Ca2+ level of HEK293 cell culture media after transfection increases the success of functional insect OR expression.

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 Dates: 2018-11-222018-11-242018
 Publication Status: Published in print
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 Identifiers: Other: HAN318
DOI: 10.1016/j.jneumeth.2018.11.021
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Title: Journal of Neuroscience Methods
  Other : J. Neurosci. Meth.
Source Genre: Journal
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Publ. Info: Amsterdam : Elsevier
Pages: - Volume / Issue: 312 Sequence Number: - Start / End Page: 122 - 125 Identifier: ISSN: 0165-0270
CoNE: https://pure.mpg.de/cone/journals/resource/954925480594