Formalin-induced short- and long-term modulation of Cav currents expressed in Xenopus oocytes: an in vitro cellular model for formalin-induced pain.
Xenopus oocytes expressing high voltage-gated calcium channels (Ca(v)) were exposed to formalin (0.5%, v/v, 5 min.) and the oocyte death and Ca(v) currents were studied for up to 10 days. Ca(v) channels were expressed with alpha(1)beta(1)b and alpha(2)delta sub-units and the currents (I(Ba)) were studied by voltage clamp. None of the oocytes was dead during the exposure to formalin. Oocyte death was significant between day 1 and day 5 after the exposure to formalin and was uniform among the oocytes expressing various Ca(v) channels. Peak I(Ba) of all Ca(v) and A(1), the inactivating current component was decreased whereas the non-inactivated R current was not affected by 5 min. exposure to formalin. On day 1 after the exposure to formalin, Ca(v)1.2c currents were increased, 2.1 and 2.2 currents were decreased and 2.3 currents were unaltered. On day 5, both peak I(Ba) and A(1) currents were increased. Ca(v)1.2c, 2.2 and 2.3 currents were increased and Ca(v)2.1 was unaltered on day 10 after the exposure to formalin. Protein kinase C (PKC) may be involved in formalin-induced increase in Ca(v) currents due to the (i) requirement for Ca(v)beta(1)b sub-units; (ii) decreased phorbol-12-myristate,13-acetate potentiation of Ca(v)2.3 currents; (iii) absence of potentiation of Ca(v)2.3 currents following down-regulation of PKC; and (iv) absence of potentiation of Ca(v)2.2 or 2.3 currents with Ser-->Ala mutation of Ca(v)alpha(1)2.2 or 2.3 sub-units. Increased Ca(v) currents and PKC activation may coincide with changes observed in in vivo pain investigations, and oocytes incubated with formalin may serve as an in vitro model for some cellular mechanisms of pain.
Rajagopal
Senthilkumar
S
Department of Anesthesiology, University of Virginia, Charlottesville, VA, USA.
Fang
Hongyu
H
Lynch
Carl
C
Kamatchi
Ganesan L
GL
D000818:Animals; D015220:Calcium Channels / Q000187:drug effects* / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D004195:Disease Models, Animal; D005260:Female; D005557:Formaldehyde / Q000633:toxicity*; D009865:Oocytes / Q000187:drug effects* / Q000473:pathology; D010146:Pain / Q000139:chemically induced*; D018408:Patch-Clamp Techniques; D011493:Protein Kinase C / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D013997:Time Factors; D014982:Xenopus laevis
D015220:Calcium Channels; D005557:Formaldehyde; D011493:Protein Kinase C