title: Pluronic F-127 affects the regulation of cytoplasmic Ca2+ in neuronal cells.
abstract: Fura-2 is one of the most widely used cytoplasmic Ca2+ ([Ca2+]cyt) sensors. In studies using isolated dorsal root ganglion (DRG) neurons, the loading of Fura-2 AM is often facilitated by the use of pluronic F-127. In preliminary studies, we detected that the use of pluronic F-127 appeared to be affecting the depolarization-evoked [Ca2+]cyt transient in DRG neurons. To determine whether this was the case, we conducted a systematic study. Adult rat DRG neurons were cultured, and their response to 50 mM KCl was measured in sister cultured cells (isolated on the same day) that were loaded with 5 microM Fura-2AM in the absence or in the presence of 0.02% pluronic F-127. In the absence of pluronic F-127, the KCl-evoked [Ca2+]cyt transient changed with time, being wider on day 1 than on day 2 after plating. On day 2, the KCl-evoked [Ca2+]cyt transient was wider in neurons Fura-2 loaded in the presence of pluronic F-127. These results indicate that pluronic F-127 significantly alters depolarization-evoked [Ca2+]cyt transients, which may reflect alteration in regulation of [Ca2+]cyt in neuronal cells.
authors.affiliation: Department of Anesthesiology, New York University School of Medicine, 550 First Avenue, RR605, New York, NY 10016, USA.
authors.forename: Jhon-Jairo
authors.initials: JJ
authors.lastname: Sutachan
authors.forename: José V
authors.initials: JV
authors.lastname: Montoya G
authors.forename: Fang
authors.initials: F
authors.lastname: Xu
authors.forename: Daniel
authors.initials: D
authors.lastname: Chen
authors.forename: Thomas J J
authors.initials: TJ
authors.lastname: Blanck
authors.forename: Esperanza
authors.initials: E
authors.lastname: Recio-Pinto
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002448:Cell Adhesion; D002478:Cells, Cultured; D003593:Cytoplasm / Q000187:drug effects / Q000378:metabolism*; D005456:Fluorescent Dyes; D016257:Fura-2; D005727:Ganglia, Spinal / Q000166:cytology / Q000187:drug effects; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D020442:Poloxamer / Q000494:pharmacology*; D011189:Potassium Chloride / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013501:Surface-Active Agents / Q000494:pharmacology*; D013696:Temperature
chemical_list: D005456:Fluorescent Dyes; D013501:Surface-Active Agents; D020442:Poloxamer; D011189:Potassium Chloride; D002118:Calcium; D016257:Fura-2