Expression of P2X purinoceptors in PC12 phaeochromocytoma cells.
1. The PC12 cell line, which was cloned from a rat adrenal phaeochromocytoma, is a useful model system. It expresses neuronal properties after treatment with nerve growth factor (NGF). The nervous system-specific P2X receptor subtype P2X(2) was initially cloned from PC12 cells, but little is known about the expression of other subtypes of P2X receptors in PC12 cells. The aim of the present study was to investigate whether PC12 cells express the other P2X receptors when exposed to NGF. 2. Reverse transcription-polymerase chain reaction at the mRNA level and immunocytochemisty at the protein level showed that, among the seven P2X purinoceptor subtypes, only P2X(2) was found to be expressed in undifferentiated PC12 phaeochromocytoma cells, but all seven P2X purinoceptor subtypes were expressed in differentiated PC12 cells treated with 50 microg/mL NGF. 3. Electrophysiological recordings indicated that ATP (30 micromol/L) but not alpha,beta-methylene ATP (alpha,beta-meATP; 30 micromol/L) evoked an inward current in undifferentiated PC12 cells, but both alpha,beta-meATP and ATP evoked inward currents in differentiated PC12 cells. The results indicate that the NGF-induced P2X receptors expressed in PC12 cells are functional channels. 4. The present study suggests that the NGF-induced neuronal phenotype of PC12 cells may be a model for the study of P2X heteromeric receptors.
Sun
Ji-Hu
JH
Department of Physiology, Second Military Medical University, Shanghai, China.
Cai
Guo-Jun
GJ
Xiang
Zheng-Hua
ZH
D000818:Animals; D020932:Nerve Growth Factor / Q000494:pharmacology*; D016716:PC12 Cells; D018408:Patch-Clamp Techniques; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D018048:Receptors, Purinergic P2 / Q000502:physiology*; D058469:Receptors, Purinergic P2X
D012333:RNA, Messenger; D018048:Receptors, Purinergic P2; D058469:Receptors, Purinergic P2X; D020932:Nerve Growth Factor