Receptors involved in nerve-mediated vasoconstriction in small arteries of the rat hepatic mesentery

1. We have investigated the neurotransmitters and receptor subtypes involved in nerve-mediated vasoconstriction in small arteries of the rat hepatic mesentery. 2. A dense sympathetic innervation was demonstrated antibodies against the synaptic vesicle protein synaptophysin. 3. Reverse transcription-polymerase chain reaction (RT-PCR) demonstrated very strong expression of the α(1A)-adrenergic, neuropeptide Y (NPY) Y₁, P(2X1)- and P(2X4)-purinergic receptors, moderate expression of the α(2B)-adrenergic receptor and the purinergic P(2X5)- and P(2X7)-receptors and weak expression of the α(1B)-, α(1D)-, α(2A)- and α(2C)-adrenergic receptors and the P(2X2)- and P(2X3)-purinergic receptors. NPY₂ and P(2X6) receptor expression was absent. 4. Electrical field stimulation (10 Hz, 10 s) produced contractions which were abolished by tetrodotoxin (10^-6 M) and/or guanethidine (GE, 5 x 10^-6 M) and a combination of benextramine (10^-5 M) and α,β-methylene ATP, (α,β-mATP, 3 x 10^-6 M) or PPADS (10^-5 M). Selective α₁-adrenergic receptor antagonists showed the potency order of prazosin > WB-4101 > 5-methyl-urapidil > BMY 7378. Yohimbine (10^-8 M, 10^-7 M), α,β-mATP (3 x 10^-6 M) and PPADS (10^-5 M) each enhanced the response to nerve stimulation. 5. Some experiments demonstrated a slow neurogenic contraction which was abolished by GE or the selective NPY₁ receptor antagonist 1229U91 (6 x 10^-7 M). 6. We conclude that nerve-mediated vasoconstriction results from the activation of postsynaptic α(1A)-adrenergic and P(2X)-purinergic receptors and under some conditions, NPY₁ receptors. Neurotransmitter release is modulated by presynaptic α₂-adrenergic receptors and possibly also P(2X)-purinoceptors. The major postsynaptic subtypes involved were well predicted by mRNA expression as measured by RT-PCR, suggesting that this technique may be a useful adjunct to studies aimed at identifying functional receptor subtypes.