Contribution of NK(2) tachykinin receptors to propulsion in the rabbit distal colon. (57/4453)

The role of the tachykinin neurokinin (NK)(2) receptors on rabbit distal colon propulsion was investigated by using two selective NK(2)-receptor antagonists, MEN-10627 and SR-48968. Experiments on colonic circular muscle strips showed that contractile responses to [beta-Ala(8)]NKA-(4-10) (1 nM-1 microM), a selective NK(2)-receptor agonist, were competitively antagonized by MEN-10627 (1-100 nM), whereas SR-48968 (0.1-10 nM) caused an insurmountable antagonism, thus confirming the difference in the mode of action of the two compounds. Colonic propulsion was elicited by distending a mobile rubber balloon with 0.3 ml (submaximal stimulus) or 1.0 ml (maximal stimulus) of water. The velocity of anal displacement of the balloon (mm/s) was considered the main propulsion parameter. At low concentrations (1.0-100 nM and 0.1-10 nM, respectively), MEN-10627 and SR-48968 facilitated the velocity of propulsion, whereas at high concentrations (100 nM and 1 microM, respectively) they decelerated propulsion. The excitatory and inhibitory effects of both antagonists were observed only with submaximal stimulus. We focused on the hypothesis that the facilitatory effect on propulsion may result from blockade of neuronal NK(2) receptors and the inhibitory effect from suppression of the excitatory transmission mediated by NK(2) receptors on smooth muscle cells. In the presence of N(G)-nitro-L-arginine (300 microM), a nitric oxide synthase inhibitor, MEN-10627, at a concentration (10 nM) that was found to accelerate propulsion in control experiments inhibited the velocity of propulsion. In the presence of threshold (1-10 nM) or full (1 microM) concentration of atropine, which inhibited to a great extent the velocity of propulsion, the inhibitory effect of MEN-10627 (1 microM) was markedly increased. In conclusion, in the rabbit distal colon NK(2) receptors may decelerate propulsion by activating a nitric oxide-dependent neuronal mechanism and may accelerate it by a postjunctional synergistic interaction with cholinergic muscarinic receptors.  (+info)

Prenyltransferase inhibitors block superoxide production by pulmonary vascular smooth muscle. (58/4453)

We recently showed that the farnesyltransferase inhibitor FTI-277 blocks interleukin 1beta (IL-1beta)-induced nitric oxide production in pulmonary vascular smooth muscle cells (SMC), whereas the geranylgeranyltransferase inhibitor GGTI-298 enhances this effect. Here we show that IL-1beta and platelet-derived growth factor (PDGF) stimulate superoxide production by pulmonary vascular SMC and that this effect is blocked by both FTI-277 and GGTI-298, suggesting that farnesylated and geranylgeranylated proteins are required for superoxide production. We also show that FTI-277 and GGTI-298 block superoxide production stimulated by constitutively active mutant H-Ras. Furthermore, superoxide production by IL-1beta, PDGF factor, and constitutively activated Ras is blocked by diphenyleneiodonium, implicating NAD(P)H oxidase as the generating enzyme. Given the role of oxidant radicals in vascular reactivity and injury, the action of both FTI-277 and GGTI-298 in suppressing superoxide generation by an inflammatory cytokine as well as by a potent smooth muscle mitogen may be therapeutically useful.  (+info)

Effects of intracolonic opioid receptor agonists on polymodal pelvic nerve afferent fibers in the rat. (59/4453)

We studied the effects of intracolonic administration of opioid receptor agonists (ORAs) on responses of pelvic nerve afferent fibers to colorectal distension (CRD) and heat. Single-fiber recordings were made from the decentralized S1 dorsal rootlet in the rat. An approximately 7-cm length of descending colon was isolated in situ to permit intracolonic perfusion with Krebs solution, which, when the outflow was clamped, was used to distend the colon. Responses to noxious CRD (40 mmHg, 30 s) were tested after intracolonic instillation of mu-, delta- or kappa-ORAs. Intracolonic administration of the kappa-ORAs EMD 61,753 (n = 5/12) and U62,066 (n = 8/11), but not either the mu-ORA fentanyl or the delta-ORA SNC-80, concentration-dependently inhibited responses of afferent fibers. For fibers unaffected by intracolonic administration of EMD 61,753 or U62,066, intra-arterial administration of kappa-ORAs was effective. Forty-one of 54 mechanosensitive fibers also responded to intracolonic instillation of heated Krebs solution (50 degrees C). Intra-arterial injection of fentanyl or SNC-80 did not attenuate responses to heat. Either intracolonic or intra-arterial administration of EMD 61,753 or U62, 066, however, inhibited afferent fiber responses to heat. These results document that mechanical and thermal sensitivity of polymodal pelvic nerve afferent fibers innervating the rat colon can be inhibited peripherally by intracolonic instillation of kappa-ORAs.  (+info)

5-HT1P receptor-mediated slow depolarization in neurons of guinea pig inferior mesenteric ganglion. (60/4453)

AIM: To study the effects of several 5-hydroxytryptamine (5-HT) receptor subtype antagonists on 5-HT-induced depolarization and the effects of 5-HT1P receptor agonist on the membrane potential in the neurons of guinea pig inferior mesenteric ganglion (IMG). METHODS: Intracellular recordings were made from neurons of the isolated guinea pig IMG. RESULTS: Cyproheptadine (5-HT1/2 antagonist 10 mumol.L-1, n = 7) and BRL 24924 (5-HT1P antagonist 10 mumol.L-1, n = 19) reversibly suppressed 5-HT slow response; pressure ejection of MCPP (5-HT1P agonist 10 mmol.L-1) induced a slow depolarization in most of 5-HT sensitive neurons (10/14). CONCLUSION: 5-HT-induced slow depolarization is mediated by 5-HT1P receptor.  (+info)

Optimization of automated quantification of 123I-IBZM uptake in the striatum applied to parkinsonism. (61/4453)

Evaluation of therapies for parkinsonism by dopamine receptor SPECT requires a reproducible, optimized quantitation technique. This study presents a new, objective, automated technique for semiquantitative analysis of dopamine receptor density, as applied to the differential diagnosis of parkinsonism. METHODS: Dopamine receptor density measured by 123I-iodobenzamide (IBZM) SPECT was retrospectively analyzed in nonidiopathic parkinsonism (NIPS), in Parkinson's disease (PD), and in healthy volunteers (n = 19, 38, and 13, respectively). A mean template was created from coregistered control studies. Registration errors were assessed using studies with simulated binding deficits. Patient studies were registered to the mean template, and striatal binding was calculated from a corresponding map of 3-dimensional regions of interest (ROIs). The striatal binding ratio and deficits determined by voxelwise comparison with the normal template were investigated and tested with various 3-dimensional ROI sizes and positions. Separation of patient groups was determined by tscore after automatically processing all studies. Results were compared with manual ROI analyses. RESULTS: The automatic method was completely reproducible in 64 of 70 cases. The best diagnostic discriminator was the minimum binding ratio of the 2 striatal nuclei, with the following values: NIPS, 1.33+/-0.13; PD, 1.50+/-0.12; healthy volunteers, 1.49+/-0.08 (+/-SD). The deficit size from voxelwise analysis was: NIPS, 20.5+/-8.2 mL; PD, 9.5+/-8.3; healthy volunteers, 8.9+/-6.0 (+/-SD). The accuracy, measured by receiver operating characteristic areas, was 0.85+/-0.05, 0.77+/-0.06, and 0.80+/-0.06 (+/-SE) for the optimal predictor (automated) and 2 blinded observers (manual), respectively. CONCLUSION: A new 3-dimensional, automated technique has been developed to semiquantitate receptor density that dramatically improves reproducibility. The optimal diagnostic discriminator of parkinsonism determined by the automatic technique has good accuracy compared with the manual technique.  (+info)

Endothelium-dependent relaxation followed by contraction mediated by NK(1) receptors in precontracted rabbit intrapulmonary arteries. (62/4453)

In the present study, we examined whether substance P (SP) and SP methyl ester (SPME), a selective NK(1) agonist, cause biphasic responses consisting of endothelium-dependent relaxation (EDR) and contraction (EDC) in precontracted rabbit intrapulmonary arteries. In arteries contracted with PGF(2alpha) (2x10(-6) M), SP as well as SPME caused only EDR at low concentration (10(-9) M) and EDR followed by EDC at higher concentrations, indicating the involvement of NK(1) receptors. The SP (10(-8) M)-induced EDR was abolished in arteries moderately contracted by PGF(2alpha) (5x10(-7) M) and the EDC in arteries maximally contracted by PGF(2alpha) (10(-5) M), indicating that EDR and EDC are inversely dependent on preexisting tone. Indomethacin (10(-8) - 10(-6) M), a cyclo-oxygenase inhibitor, and ozagrel (10(-8) - 10(-6) M), a TXA(2) synthetase inhibitor attenuated the EDC in the SPME (10(-7) M)-induced biphasic response and markedly potentiated the EDR. AA-861 (10(-8) - 10(-6) M), a 5-lipoxygenase inhibitor, did not affect the EDR or EDC. L-N(G)-nitro-arginine methyl ester (10(-5) - 10(-4) M), a nitric oxide synthase inhibitor, attenuated the EDR and slightly potentiated the EDC. CP-99994 (10(-10) - 10(-8) M), an NK(1) antagonist, attenuated the EDC and potentiated the EDR in the SPME (10(-7) M)-induced biphasic response, while the NK(2) antagonist SR-48968 (10(-9) - 10(-7) M) had no effect. CP-99994 attenuated the SPME (10(-7) M)-induced EDC under EDR-blockade to a greater extent than the EDR under EDC-blockade, indicating that CP-99994 enhanced the EDR component by preferential inhibition of the EDC component. In conclusion, NK(1) agonists caused a biphasic endothelium-dependent response (EDR and EDC) in submaximally precontracted intrapulmonary arteries. The EDC and EDR mediated by NK(1) receptors may play physiological and/or pathophysiological roles in modulation of vascular tone.  (+info)

Respiratory actions of tachykinins in the nucleus of the solitary tract: characterization of receptors using selective agonists and antagonists. (63/4453)

1. The respiratory response to microinjection of tachykinins and analogues into the commissural nucleus of the solitary tract (cNTS) of urethane-anaesthetized rats was investigated in the presence and absence of selective tachykinin NK(1), NK(2) and NK(3) antagonists (RP 67580, SR 48968 and SR 142801, respectively). 2. All tachykinins, except for the selective NK(2) agonist, [Nle(10)]-NKA(4-10), increased tidal volume (VT). The rank potency order of naturally-occurring tachykinins was neurokinin A (NKA)> or =substance P (SP)>>NKB, whereas the rank order for selective analogues was senktide> or = septide>> [Sar(9),Met(O(2))(11)]-SP>>[Nle(10)]-NKA(4-10). Septide (NK(1)-selective) and senktide (NK(3)-selective) were 22 fold more potent (pD(2) approximately 12) at stimulating VT than SP (pD(2) approximately 10.5). 3. Tachykinin agonists produced varying degrees of respiratory slowing, independent of changes in VT. At doses producing maximum stimulation of VT, agonists induced either a mild (<10 breaths min(-1) decrease; SP and septide), moderate (10 - 25 breaths min(-1) decrease; NKA, NKB and [Sar(9),Met(O(2)]-SP) or severe ( approximately 40 breaths min(-1) decrease; senktide) bradypnoea. [Nle(10)]-NKA(4-10) produced a dose-dependent bradypnoea without affecting VT. 4. RP 67580 significantly attenuated the VT response to SP (33 pmol) and NKA (10 pmol) but not NKB (100 pmol). In the presence of RP 67580, the mild bradypnoeic response to NKB was significantly enhanced whereas SP and NKA induced a bradyapnea which was not observed in the absence of RP 67580. SR 48968 had no effect on the VT response to SP or NKB, markedly enhanced the VT response to NKA and completely blocked the bradypnoeic response to [Nle(10)]-NKA(4-10). Only SR142801 attenuated the VT response to NKB. 5. The present data suggest that all three tachykinin receptors (NK(1), NK(2) and NK(3)) are present in the cNTS and are involved in the central control of respiration.  (+info)

Antisense inhibition of delta-opioid receptor gene function in vivo by peptide nucleic acids. (64/4453)

Peptide nucleic acids (PNA) are synthetic analogs of DNA that hybridize to complementary oligonucleotide sequences with exceptional affinity and target specificity. The stability of PNA in biological fluids together with the unique hybridization characteristics of these structures suggests that PNA may have considerable potential as antisense agents for experimental use in vivo. To test this hypothesis, we attempted to modulate supraspinal delta-opioid receptor function in rats using PNA sequences designed to be complementary to a region of the rat delta-opioid receptor. Repeated i.c.v. administration of PNA over a period of 5 days significantly inhibited the antinociceptive response and locomotor response to selective delta-opioid receptor agonists. PNA attenuated delta-opioid receptor function in a sequence-specific, target-specific, and reversible manner characteristic of the functional inhibition caused by an antisense mechanism. There were no apparent toxicities arising from the PNA treatment based on the behavior of the animals and inspection of the treated tissues. Saturation binding studies on brain homogenates did not reveal any significant difference in receptor B(max) between treatment groups. However, [(35)S]guanosine-5'-O-(3-thio)triphosphate binding assays demonstrated a significant decrease in agonist efficacy in homogenates prepared from antisense-treated rats. Taken together, these results demonstrate that peptide nucleic acids are effective antisense agents in vivo and suggest that PNA may be a useful alternative to phosphodiester or phosphorothioate oligonucleotides, or variants thereof, for determination of gene function in vivo.  (+info)