Muscarinic cholinergic regulation of L-type calcium channel in heart of embryonic mice at different developmental stages

AIM: To investigate the muscarinic regulation of L-type calcium current (I(Ca-L)) during development. METHODS: The whole cell patch-clamp technique was used to record I(Ca-L) in mice embryonic cardiomyocytes at different stages (the early developmental stage, EDS; the intermediate developmental stage, IDS; and the late developmental stage, LDS). Carbachol (CCh) was used to stimulate M-receptor in the embryonic cardiomyocytes of mice. RESULTS: The expression Of ICa-L density did not change in different developmental stages (P>0.05). There was no difference in the sensitivity Of ICa-L to CCh during development (P>0.05). This inhibitory action of CCh was mediated by inhibition of cyclic AMP since 8-bromo-cAMP completely reversed the muscarinic inhibitory action. IBMX, a non-selective inhibitor of phosphodiesterase (PDE), reversed the inhibitory action of M-receptor on I(Ca-L) current by 71.2 %+/-9.2 % (n=8) and 11.3 %+/-2.5 % (n=9) in EDS and LDS respectively. However forskolin, an agonist of adenylyl cyclase (AC), reversed the action of CCh by 14.5 %+/-3.5 % (n=5) and 82.7 %+/-10.4 % (n=7) in EDS and LDS respectively. CONCLUSION: The inhibitory action of CCh on I(Ca-L) current was mediated in different pathways: in EDS, the inhibitory action of M-receptor on I(Ca-L) channel mainly depended on the stimulation of PDE. However, in LDS, the regulation by M-receptor on I(Ca-L) channel mainly depended on the inactivation of AC.