| Autor: |
Joshi PR; School of Pharmacy, The University of Louisiana at Monroe, Monroe, LA 71209, USA., Suryanarayanan A, Schulte MK |
| Jazyk: |
angličtina |
| Zdroj: |
Journal of neuroscience methods [J Neurosci Methods] 2004 Jan 15; Vol. 132 (1), pp. 69-79. |
| DOI: |
10.1016/j.jneumeth.2003.09.002 |
| Abstrakt: |
Xenopus laevis oocytes are used extensively in the study of ion channel coupled receptors. Efficient use of oocytes for ion channel characterization requires a system that is inherently stable, reproducible, minimizes drug volumes, and maximizes oocyte longevity. We have constructed a vertical flow oocyte recording chamber to address the aforesaid issues, where the oocyte is placed in a funnel-shaped chamber and perfused from the bottom of the funnel. The vertical rather than horizontal flow of perfusate results in an unusually stable environment for oocyte recording. Two-electrode voltage clamp recordings from a single oocyte are acquired easily and routinely over several hours while maintaining stable baseline currents and reproducible response profiles. Chamber characteristics were tested using a serotonin ligand-gated ion channel receptor (5-HT3R). Data obtained from this system corresponds well with published data. To further test the stability and reliability of this perfusion chamber, we constructed an automated oocyte perfusion system utilizing a commonly available HPLC autosampler. We were able to obtain dose-response curves for various 5-HT3AR ligands using the automated perfusion system with minimal user intervention. Such a system can easily satisfy need for automated oocyte electrophysiology in academic settings, especially small to medium sized laboratories. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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