Abstract
Nonspecific binding of anandamide to plastic exhibits many features that could be mistaken as biological processes, thereby representing an important source of conflicting data on the uptake and release of this lipophilic substance. Herein, we propose an improved method to assay anandamide transport, by using glass slides (i.e., coverslips) as physical support to grow cells. Although the results obtained using plastic do not differ significantly from those obtained using glass, the new procedure has the advantage of being faster, simpler, and more accurate. In fact, the lack of aspecific adsorption of anandamide to the glass surface yields a lower background and a higher precision and accuracy in determining transport kinetics, especially for the export process. Remarkably, the kinetic parameters of anandamide uptake obtained with the old and the new procedures may be similar or different depending on the cell type, thus demonstrating the complexity of the interference of plastic on the transport process.jlr In addition, the novel procedure is particularly suitable for visualization and measurement of anandamide transport in intact cells by using a biotinylated derivative in confocal fluorescence microscopy.
| Original language | English |
|---|---|
| Pages (from-to) | 2435-2444 |
| Number of pages | 10 |
| Journal | Journal of Lipid Research |
| Volume | 51 |
| Issue number | 8 |
| DOIs | |
| Publication status | Published - Aug 2010 |
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Keywords
- Biotinylated derivative of anandamide
- Confocal microscopy
- Endocannabinoids
- Glass coverslips
- Lipid efflux
- Plasticware
ASJC Scopus subject areas
- Biochemistry
- Cell Biology
- Endocrinology
- Medicine(all)
Cite this
Pitfalls and solutions in assaying anandamide transport in cells. / Oddi, Sergio; Fezza, Filomena; Catanzaro, Giuseppina; De Simone, Chiara; Pucci, Mariangela; Piomelli, Daniele; Finazzi-Agrò, Alessandro; Maccarrone, Mauro.
In: Journal of Lipid Research, Vol. 51, No. 8, 08.2010, p. 2435-2444.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Pitfalls and solutions in assaying anandamide transport in cells
AU - Oddi, Sergio
AU - Fezza, Filomena
AU - Catanzaro, Giuseppina
AU - De Simone, Chiara
AU - Pucci, Mariangela
AU - Piomelli, Daniele
AU - Finazzi-Agrò, Alessandro
AU - Maccarrone, Mauro
PY - 2010/8
Y1 - 2010/8
N2 - Nonspecific binding of anandamide to plastic exhibits many features that could be mistaken as biological processes, thereby representing an important source of conflicting data on the uptake and release of this lipophilic substance. Herein, we propose an improved method to assay anandamide transport, by using glass slides (i.e., coverslips) as physical support to grow cells. Although the results obtained using plastic do not differ significantly from those obtained using glass, the new procedure has the advantage of being faster, simpler, and more accurate. In fact, the lack of aspecific adsorption of anandamide to the glass surface yields a lower background and a higher precision and accuracy in determining transport kinetics, especially for the export process. Remarkably, the kinetic parameters of anandamide uptake obtained with the old and the new procedures may be similar or different depending on the cell type, thus demonstrating the complexity of the interference of plastic on the transport process.jlr In addition, the novel procedure is particularly suitable for visualization and measurement of anandamide transport in intact cells by using a biotinylated derivative in confocal fluorescence microscopy.
AB - Nonspecific binding of anandamide to plastic exhibits many features that could be mistaken as biological processes, thereby representing an important source of conflicting data on the uptake and release of this lipophilic substance. Herein, we propose an improved method to assay anandamide transport, by using glass slides (i.e., coverslips) as physical support to grow cells. Although the results obtained using plastic do not differ significantly from those obtained using glass, the new procedure has the advantage of being faster, simpler, and more accurate. In fact, the lack of aspecific adsorption of anandamide to the glass surface yields a lower background and a higher precision and accuracy in determining transport kinetics, especially for the export process. Remarkably, the kinetic parameters of anandamide uptake obtained with the old and the new procedures may be similar or different depending on the cell type, thus demonstrating the complexity of the interference of plastic on the transport process.jlr In addition, the novel procedure is particularly suitable for visualization and measurement of anandamide transport in intact cells by using a biotinylated derivative in confocal fluorescence microscopy.
KW - Biotinylated derivative of anandamide
KW - Confocal microscopy
KW - Endocannabinoids
KW - Glass coverslips
KW - Lipid efflux
KW - Plasticware
UR - http://www.scopus.com/inward/record.url?scp=77956830425&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77956830425&partnerID=8YFLogxK
U2 - 10.1194/jlr.D004176
DO - 10.1194/jlr.D004176
M3 - Article
C2 - 20447929
AN - SCOPUS:77956830425
VL - 51
SP - 2435
EP - 2444
JO - Journal of Lipid Research
JF - Journal of Lipid Research
SN - 0022-2275
IS - 8
ER -