title: A vertical flow chamber for Xenopus oocyte electrophysiology and automated drug screening.
abstract: 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.
authors.affiliation: School of Pharmacy, The University of Louisiana at Monroe, Monroe, LA 71209, USA.
authors.forename: P R
authors.initials: PR
authors.lastname: Joshi
authors.forename: A
authors.initials: A
authors.lastname: Suryanarayanan
authors.forename: M K
authors.initials: MK
authors.lastname: Schulte
mesh_terms: D000818:Animals; D001331:Automation / Q000295:instrumentation* / Q000379:methods; D017076:Computer-Aided Design; D015194:Diffusion Chambers, Culture / Q000295:instrumentation* / Q000379:methods; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004553:Electric Conductivity; D004594:Electrophysiology; D005260:Female; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D008845:Microinjections / Q000379:methods; D009865:Oocytes / Q000187:drug effects* / Q000502:physiology; D018408:Patch-Clamp Techniques; D010477:Perfusion / Q000379:methods; D018075:RNA, Complementary / Q000096:biosynthesis; D044406:Receptors, Serotonin, 5-HT3 / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D012701:Serotonin / Q000494:pharmacology; D014982:Xenopus laevis
chemical_list: D018075:RNA, Complementary; D044406:Receptors, Serotonin, 5-HT3; D012701:Serotonin