Objective: Our major hypothesis is that cyclic adenosine-3',5'-monophosphate (cAMP)-mediated modulation of neurotransmitter release plays different roles at low and high activity of the sympathetic nervous system. We further hypothesize that cAMP-mediated neuromodulation might underlie disparate neurovascular control in mesenteric arteries and veins.
Methods: Electrical field stimulation (EFS)-evoked overflow of noradrenaline (NA) was evaluated in the absence or presence of activators and inhibitors of cAMP-dependent pathway at low (4 Hz) and high (16 Hz) frequencies of stimulation of endothelium-denuded secondary and tertiary branches of the canine isolated inferior mesenteric arteries and veins. The content of NA in samples of the superfusates collected before and during nerve stimulation was assayed by high-performance liquid chromatography (HPLC) technique in conjunction with electrochemical detection. Student's t-test and ANOVA analyses were applied for statistical analysis.
Results: Activation of cAMP-dependent pathway with either isoproterenol (ISO, 10 microM), forskolin (1 microM), dibutyryl cAMP (100 microM) or combined site-specific activators of cAMP-dependent protein kinase (PKA) [i.e. N(6)-phenyl-adenosine-3',5'-cyclic monophosphate, 8-(6-aminohexyl) aminoadenosine-3',5'-cyclic monophosphate, and the Sp-isomer of 5,6-dichloro-1-D-ribofuranosylbenzimidazole-3',5'-cyclic monophosphorothioate, each 100 microM] caused an enhancement of the EFS-evoked overflow of endogenous NA at 16 Hz of stimulation but was without an effect at 4 Hz of stimulation both in artery and vein. The EFS (16 Hz)-evoked overflow of NA in vein was also increased in the presence of inhibitors of phosphodiesterase (PDE) III and PDE IV (i.e. milrinone, 0.4 microM, and roilpram, 30 microM), whereas these inhibitors did not affect the overflow of NA in the artery. The facilitating effect of activators of cAMP-dependent pathway on the EFS-evoked release of NA at 16 Hz appears to be more pronounced in the vein than in artery. The increasing effect of ISO (10 microM) was inhibited with either propranolol (1 microM) or the adenylyl cyclase (AC) inhibitor [9-(tetrahydro-2'-furyl)adenine] (SQ 22,536, 100 microM) in both blood vessels. The ISO effect was inhibited by the PKA inhibitor 14-22 amide (PKI(14-22)), 1 microM, in the artery but not in vein. The enhancing effect of FSK was inhibited by pretreatment of the tissue with SQ 22,536, 100 microM, or the PKA inhibitors PKI(14-22), 1 microM, and 4-cyano-3-methylisoquinoline, 50 nM. However, the inhibitors alone did not significantly change the EFS-evoked overflow of NA in both blood vessels.
Conclusions: Activation of AC-cAMP-PKA pathway appears to play a role in modulating NA release at higher stimulation frequencies as might be expected during stress, strenuous exercise, or hemorrhage. The AC-cAMP pathway plays a more pronounced role in the autonomic neural control of mesenteric veins than of the corresponding arteries, whereas the PKA contribution is more distinct in the arteries.