TY - JOUR
T1 - Gs cascade regulates canonical transient receptor potential 5 (TRPC5) through cAMP mediated intracellular Ca 2+ release and ion channel trafficking
AU - Hong, Chansik
AU - Kim, Jinsung
AU - Jeon, Jae Pyo
AU - Wie, Jinhong
AU - Kwak, Misun
AU - Ha, Kotdaji
AU - Kim, Hana
AU - Myeong, Jongyun
AU - Kim, Sung Young
AU - Jeon, Ju Hong
AU - So, Insuk
PY - 2012/4/27
Y1 - 2012/4/27
N2 - Canonical transient receptor potential (TRPC) channels are Ca 2+-permeable, non-selective cation channels those are widely expressed in mammalian cells. Various molecules have been found to regulate TRPC both in vivo and in vitro, but it is unclear how heterotrimeric G proteins transmit external stimuli to regulate the activity of TRPC5. Here, we demonstrated that TRPC5 was potentiated by the Gα s regulatory pathway. Whole-cell TRPC5 current was significantly increased by β-adrenergic receptor agonist, isoproterenol (ISO, 246±36%, n=6), an activator of the adenylate cyclase, forskolin (FSK, 273±6%, n=5), or a membrane permeable cAMP analogue, 8-Br-cAMP (251±63%, n=7). In addition, robust Ca 2+ transient induced by isoproterenol was observed utilizing a Ca 2+ imaging technique. When intracellular [Ca 2+] i was buffered to 50nM, cAMP-induced potentiation was attenuated. We also found that the Ca 2+ release is mediated by IP 3 since intracellular IP 3 infusion attenuated the potentiation of TRPC5 by Gα s cascade. Finally, we identified that the membrane localization of TRPC5 was significantly increased by ISO (155±17%, n=3), FSK (172±39%, n=3) or 8-Br-cAMP (216±59%, n=3). In conclusion, these results suggest that the Gα s-cAMP pathway potentiates the activity of TRPC5 via facilitating intracellular Ca 2+ dynamics and increasing channel trafficking to the plasma membrane.
AB - Canonical transient receptor potential (TRPC) channels are Ca 2+-permeable, non-selective cation channels those are widely expressed in mammalian cells. Various molecules have been found to regulate TRPC both in vivo and in vitro, but it is unclear how heterotrimeric G proteins transmit external stimuli to regulate the activity of TRPC5. Here, we demonstrated that TRPC5 was potentiated by the Gα s regulatory pathway. Whole-cell TRPC5 current was significantly increased by β-adrenergic receptor agonist, isoproterenol (ISO, 246±36%, n=6), an activator of the adenylate cyclase, forskolin (FSK, 273±6%, n=5), or a membrane permeable cAMP analogue, 8-Br-cAMP (251±63%, n=7). In addition, robust Ca 2+ transient induced by isoproterenol was observed utilizing a Ca 2+ imaging technique. When intracellular [Ca 2+] i was buffered to 50nM, cAMP-induced potentiation was attenuated. We also found that the Ca 2+ release is mediated by IP 3 since intracellular IP 3 infusion attenuated the potentiation of TRPC5 by Gα s cascade. Finally, we identified that the membrane localization of TRPC5 was significantly increased by ISO (155±17%, n=3), FSK (172±39%, n=3) or 8-Br-cAMP (216±59%, n=3). In conclusion, these results suggest that the Gα s-cAMP pathway potentiates the activity of TRPC5 via facilitating intracellular Ca 2+ dynamics and increasing channel trafficking to the plasma membrane.
KW - Beta-adrenergic receptor
KW - Ca
KW - CAMP
KW - Gs
KW - Isoproterenol
KW - TRPC
UR - http://www.scopus.com/inward/record.url?scp=84860322598&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2012.03.123
DO - 10.1016/j.bbrc.2012.03.123
M3 - Article
C2 - 22490661
AN - SCOPUS:84860322598
SN - 0006-291X
VL - 421
SP - 105
EP - 111
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -