PubMed Health⌕ Search

Biomedical subjects

J E Wikberg

Publications and source records attributed to J E Wikberg.

At least 109 records · Page 6Linked to original sources
PubMed1993

Further evidence for the existence of two forms of alpha 2B-adrenoceptors in rat.

Analysis of saturation isotherms of the novel alpha 2-adrenoceptor antagonist radioligand [3H]-MK 912 revealed that the ligand labelled a homogenous population of alpha 2B-adrenoceptors in the neonatal rat lung with a Kd of 0.77 nM and a Bmax of 231 fmol/mg protein. In rat kidney, combined saturation and competition experiments, using [3H]-MK 912 and the alpha 2A-adrenoceptor selective drug guanfacine, revealed that approximately 81% of the sites labelled by [3H]-MK 912 were alpha 2B-adrenoceptors and approximately 19% alpha 2A-adrenoceptors; the Kds of [3H]-MK 912 being 1.1 and 2.0 nM and the Bmax 134 and 33 fmol/mg protein, respectively. The kidney alpha 2B-adrenoceptors were studied separately by using approximately 1.5 nM [3H]-MK 912 in the presence of 0.32 microM guanfacine, the latter which blocked ligand binding to alpha 2A-adrenoceptors completely. Analysis of drug competition curves obtained during these conditions revealed that 18 out of 20 different agonists and antagonists yielded steep and uniphasic competition curves which modelled best into one site fits. However, both guanoxabenz and LT 11 appeared to inhibit [3H]-MK 912 binding at two sites; the Kds of guanoxabenz differing approximately 120-fold and that of LT 11 differing approximately 35-fold for the two sites. Moreover, the addition of mutual fixed concentrations of either 20 microM guanoxabenz or 20 microM LT 11 completely prevented the binding of the other compound to its high affinity site, indicating that both compounds labelled the same site with the high affinity. The analysis indicated that 29% of the sites were of high and 71% of low affinity. However, in the neonatal rat lung guanoxabenz and LT 11 (as well as 15 other compounds) yielded competition curves which modelled only into one site fits. The Kds obtained in the lung correlated well with the Kds obtained in the kidney for alpha 2B-adrenoceptors; for guanoxabenz and LT 11 the values from the lung were close to those determined in the kidney for the low affinity site for guanoxabenz and LT 11. Moreover, when the rat RNG alpha 2B-adrenoceptor was expressed in COS-7 cells and its binding properties tested using [3H]-MK 912 binding, guanoxabenz, LT 11 as well as a number of other drugs inhibited the ligand binding at a single alpha 2-adrenoceptor site; the drug Kds being practically the same as those found for the neonatal rat lung. It is suggested that rat alpha 2B-adrenoceptors may exist in two forms: alpha 2B1 and alpha 2B2.(ABSTRACT TRUNCATED AT 400 WORDS)

Adrenergic alpha-Antagonists↗
PubMed1993

Detomidine binding to guinea pig liver imidazoline receptors (I-receptors) shows marked positive cooperativity.

The binding of [3H]-idazoxan to guinea pig liver membranes was measured in the presence of 3 microM rauwolscine, which prevented the binding [3H]-idazoxan to alpha 2-adrenergic receptors. Under these conditions the radioligand bound to saturable imidazoline receptors (I-receptors) with a Kd of 18 nM and a Bmax of 665 fmol mg-1 protein. Six drugs which were used to compete for [3H]-idazoxan in the liver caused competition curves of widely varying steepness. Fitting the competition curves to the standard four parameter logistic function showed that the Hill coefficients (nH) varied from 2.02 (detomidine) to 0.43 (UK-14,304), The nH's obtained in liver for the six compounds correlated strongly (r = 0.99; P < 0.001) with the corresponding nH's obtained in a previous study on the guinea pig kidney where the drugs were also tested in competition with [3H]-idazoxan (Wikberg et al. 1992). Good correlation was also found for the Log(Ki) values of drugs determined in the two tissues (r = 0.96; P < 0.005). Whereas the standard logistic function accurately described the competition curves of the 5 drugs tested in the liver for which the competition curve Hill coefficients varied between 0.43 to 1.41 (UK-14,304, rilmenidine, histamine and d- and l-medetomidine), it did not accurately fit the detomidine competition curves. Instead the detomidine competition curves could be more accurately described by a model composed of the sum of two independent logistic functions.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Molecular cloning and expression of the human melanocyte stimulating hormone receptor cDNA.

Melanocytes and melanoma cells are known to possess receptors for melanocyte stimulating hormone (MSH). A cDNA clone, designated 11D, has been isolated from human melanoma cells and encodes a MSH receptor. The cloned cDNA encodes a 317 amino acid protein with transmembrane topography characteristics of a G-protein-coupled receptor, but it does not show striking similarity to already published sequences of other G-protein-coupled receptors. When 11D cDNA is expressed in COS-7 cells, it binds an 125I-labelled MSH analogue (NDP-MSH) in a specific manner. The bound ligand could be displaced by melanotropic peptides, alpha-MSH, beta-MSH, gamma-MSH and ACTH (adrenocorticotropic hormone), but not by the non-melanotropic peptide, beta-endorphin. This is the first report of the cloning of the receptor gene of the melanotropin receptor family.

Amino Acid Sequence↗
PubMed1992

Comparative in vitro study of a series of organic nitroesters: unique biphasic concentration-effect curves for glyceryl trinitrate in isolated bovine arterial smooth muscle and lack of stereoselectivity for some glyceryl trinitrate analogues.

Four different organic nitroesters, constituting a homologous series based on unbranched polyalcohols, were compared with regard to in vitro relaxation of isolated bovine mesenteric arteries contracted with 2.5 microM phenylephrine. The organic nitroesters included ethylene glycol dinitrate (EGDN), dinitratopropane (DPN), glyceryl trinitrate (GTN), and tetranitratobutane. Glyceryl trinitrate exhibited a biphasic concentration-effect relationship, with pD2 values of 11.5 +/- 0.5 and 7.2 +/- 0.2 for the high-pD2 and low-pD2 component of the relaxation curve, respectively. The high-pD2 and low-pD2 component contributed 28 and 72% of the maximal response, respectively. EGDN, DPN, and tetranitratobutane induced monophasic concentration-effect curves with pD2 values of 7.4 +/- 0.1, 7.8 +/- 0.2, and 6.9 +/- 0.6, respectively. Stereoisomeric forms of DPN and tetranitratobutane showed no difference with regard to relaxing potency in bovine mesenteric artery. GTN has a partly unique mechanism for vascular smooth muscle relaxation that distinguishes this compound from other related organic nitroesters.

Animals↗
PubMed1992

[3H]-MK 912 binding delineates two alpha 2-adrenoceptor subtypes in rat CNS one of which is identical with the cloned pA2d alpha 2-adrenoceptor.

1. Simultaneous computer modelling of control and guanfacine-masked [3H]-MK 912 saturation curves as well as guanfacine competition curves revealed that the drugs bound to two alpha 2-adrenoceptor subtypes in the rat cerebral cortex with very different selectivities. These alpha 2-adrenoceptor subtypes were designated alpha 2A and alpha 2C. The Kd value of [3H]-MK 912 for the alpha 2A-subtype was 1.77 nM and for the alpha 2C-subtype 0.075 nM; the receptor sites showing capacities 296 and 33 fmol mg-1 protein, respectively. The Kds of guanfacine were 19.9 and 344 nM, respectively. 2. Binding constants of 26 compounds for the two rat cerebral cortex alpha 2-adrenoceptor subtypes were determined by simultaneous computer modelling of control and guanfacine-masked drug competition curves as well as plain guanfacine competition curves using [3H]-MK912 as labelled ligand (i.e. a '3-curve assay'). Of the tested drugs WB4101, corynanthine, rauwolscine, yohimbine, ARC 239 and prazosin were found to be clearly alpha 2C-selective with selectivities ranging from 16 to 30 fold whereas guanfacine, oxymetazoline, BRL 44408 and BRL 41992 were found to be alpha 2A-selective with selectivities ranging from 9 to 22 fold. 3. The Kds of compounds obtained for the cerebral cortex alpha 2C-adrenoceptors showed an almost 1:1 correlation with the corresponding Kds for alpha 2-adrenoceptors expressed by the pA2d-gene (the rat 'alpha 2-C4' adrenoceptor) in CHO-cells. The cerebral cortex alpha 2A-adrenoceptors did not correlate well with the pA2d alpha 2-adrenoceptor Kds. 4. In the rat spinal cord [3H]-MK 912 bound to alpha 2A- and alpha 2C-adrenoceptor sites with similar affinities as in the cerebral cortex and with densities 172 and 7.4 fmol mg-1 protein, respectively. Drug affinities for some compounds showing major selectivity for alpha 2A- and alpha 2C-adrenoceptors were fully compatible with the notion that the spinal cord sites were alpha 2A- and alpha 2C-adrenoceptors.

Adrenergic alpha-Antagonists↗
PubMed1992

Evidence that drug binding to non-adrenergic [3H]-idazoxan binding sites (I-receptors) occurs to interacting or interconvertible affinity forms of the receptor.

Characterization of [3H]idazoxan binding to guinea pig kidney membranes showed that approximately 90% bound to nonadrenergic I-receptors and approximately 10% to alpha 2-adrenoceptors. I-Receptors could be studied separately by including 3 microM rauwolscine to the assay. During these conditions 22 different compounds out of 29 (including imidazoline and guanidinium compounds) generated biphasic or shallow competion curves (Hill coefficients down to 0.57), for which computer modelling suggested that drugs bound to two sites with different affinities. However, the proportion of sites varied considerably depending on which drug was used as competitor; the variation being from approximately 50/50% to approximately 8/92% and analysis of variance clearly indicated that the variation of proportions of sites could be attributed to an effect induced by the drugs which indicated that the sites were dynamically formed or modulated by the presence of the drug. A few drugs (guanabenz, (-)-medetomidine, phentolamine, clemastin, prazosin and idazoxan itself) yielded steep uniphasic curves (Hill coefficients near unity) which were resolved only into one site fits. One drug (detomidine) yielded supersteep competition curves (Hill coefficient 1.29), the data being reminiscent of positive cooperativity. In another set of experiments attempts were made to block one of the affinity forms of the I-receptor with histamine, a compound which had grossly different affinities for the two I-receptor sites, and competition curves were then obtained using other drugs which also seemed to distinguish between the two sites. Computer analysis of experimentally obtained as well as simulate,d computer generated, competition curves clearly showed that the kidney I-receptors did not exist in two independent non-interacting forms. Instead the data suggested that the I-receptor was composed of interacting or interconverting receptor entities with different affinities for drugs. Monovalent cations such as Cs+ or NH4+ were found to be powerful inhibitors of [3H]idazoxan binding to the kidney I-receptors, the ions decreasing both the apparent Bmax and affinity of the ligand for the receptor. The cations also decreased the affinities of UK-14,304 for both the high and low affinity forms of the I-receptor, but the apparent proportions of sites were not at all affected by the ions. Moreover, ligand binding to kidney I-receptor were not at all affected by non-hydrolyzable guanine nucleotides. It is suggested that the binding data reflects functional properties of the I-receptor protein.

Adrenergic alpha-Antagonists↗
PubMed1991

Heterogeneity in human soluble guanylate cyclase due to alternative splicing.

Two forms of the smaller subunit of the human soluble guanylate cyclase enzyme have been cloned by using PCR. One of the clones (HSGC-1) is identical to bovine and rat lung smaller subunit cyclase. However, the other (HSGC-2) is lacking 33 amino acids. Comparison of its sequence with published partial genomic sequences of bovine guanylate cyclase indicates that HSGC-2 is formed due to alternative splicing.

Animals↗
PubMed1991

Delineation of rat kidney alpha 2A- and alpha 2B-adrenoceptors with [3H]RX821002 radioligand binding: computer modelling reveals that guanfacine is an alpha 2A-selective compound.

We developed a method for the simultaneous determination of drug affinity constants for rat alpha 2A- and alpha 2B-adrenoceptor subtypes by using [3H]RX821002 radioligand binding in the kidney. Three competition curves were obtained for each drug: one for the test compound in the absence of ARC 239 (a drug found to have 108-fold higher affinity for alpha 2B- than for alpha 2A-adrenoceptors), one in the presence of ARC 239, and one for ARC 239. It is possible to determine the Kds of a tested drug for both alpha 2A- and alpha 2B-adrenoceptors by simultaneous computer modelling because of the increased constraint in the calculations given by the inclusion of ARC 239 into the assay. Using this approach, we found guanfacine and oxymetazoline to be highly alpha 2A-selective. The most alpha 2B-selective were ARC 239, prazosin and corynanthine. A number of other drugs, for example UK-14,304, rilmenidine and clonidine, were non-selective or showed minor selectivity for alpha 2A- or alpha 2B-adrenoceptors. Moreover, using Monte Carlo simulations, we showed that the three-curve method gives more accurate estimates of drug binding constants for assays when two receptor sites are present than methods analysing only one competition curve.

Animals↗
PubMed1991

Identification of an additional gene belonging to the alpha 2 adrenergic receptor family in the human genome by PCR.

We here describe the cloning of an additional gene, called alpha 2-1.8, which is similar to the previously cloned human alpha 2-adrenergic receptor located on chromosome 4. The alpha 2-1.8 gene was identified by using the polymerase chain reaction with primers specific for sequences in transmembrane regions 2 and 5 of the previously isolated human alpha 2-C4 and alpha 2-C10 adrenoceptor genes, which are localized on chromosomes 4 and 10, respectively. The new gene was identified by amplifying the 1.8 kb size fractionated region of PstI restriction cut human genomic DNA. The previously cloned alpha 2-C10 and alpha 2-C4 genes were recovered at their expected locations, 0.96 and 5.9 kb, respectively. We have identified 387 bases of the new alpha 2-1.8 gene, and its sequence is identical to the previously described alpha 2-C4 gene, but it is distinct from the alpha 2-C10 and alpha 2-C2 genes. Our results demonstrate that the alpha 2-C4 adrenergic receptor exists in more than one copy in the human genome.

Amino Acid Sequence↗
PubMed1991

Medetomidine stereoisomers delineate two closely related subtypes of idazoxan (imidazoline) I-receptors in the guinea pig.

A number of imidazoline, imidazole and guanidinium compounds and other drugs were compared for their ability to bind to non-adrenergic idazoxan (imidazoline) I-receptors in particulate guinea pig cerebral cortex and ileum smooth muscle fractions. Radioligand binding with [3H]idazoxan was used for the experiments. Computer modelling of the binding data gave dissociation constants for drug binding to both I-receptors and alpha 2-adrenoceptors. Most drugs showed similar affinities for I-receptors in cortex and ileum. However, medetomidine stereoisomers as well as a few other drugs clearly delineated the I-receptors in cortex and ileum as different.

Adrenergic alpha-Agonists↗
PubMed1991

Delineation of three pharmacological subtypes of alpha 2-adrenoceptor in the rat kidney.

1. Simultaneous computer modelling of plain and ARC 239- and guanoxabenz-masked [3H]-RX821002 saturation curves, plain ARC 239 and guanoxabenz competition curves as well as ARC 239-masked guanoxabenz competition curves revealed that the drugs bound to three alpha 2-adrenoceptor subtypes in the rat kidney with grossly differing selectivities. These alpha 2-adrenoceptor subtypes were termed alpha 2 A, alpha 2B1 and alpha 2B2. The order of affinities for [3H]-RX821002 for the adrenoceptor sites was alpha 2A greater than alpha 2B1 greater than alpha 2B2, the KdS being 0.62 +/- 0.05, 2.52 +/- 0.11 and 6.74 +/- 1.21 nM, respectively. The order of affinities for ARC 239 was alpha 2B1 greater than alpha 2B2 much greater than alpha 2A with KdS 4.78 +/- 1.04, 28.8 +/- 4.1 and 1460 +/- 270 nM, respectively. For guanoxabenz the order of affinities was alpha 2A greater than alpha 2B1 much greater than alpha 2B2 with KdS 99.7 +/- 15.1, 508 +/- 135 and 25,400 +/- 2400 nM, respectively. 2. Binding constants for 14 compounds for the three rat kidney alpha 2-adrenoceptor subtypes were determined by the simultaneous computer modelling of plain and ARC 239- and guanoxabenz-masked drug competition curves, plain ARC 239 and guanoxabenz competition curves as well as ARC 239-masked guanoxabenz competition curves. Of the 14 compounds tested, oxymetazoline and guanfacine were found to bind with low affinities to both of the alpha 2B1- and alpha 2B2-adrenoceptor but with high affinity to the alpha 2A-adrenoceptor. 3. (-)-Adrenaline and (-)-noradrenaline showed dissimilar order of affinities for the three alpha2-adrenoceptors. For (-)-adrenaline the order of affinities was alpha2Bl >- alpha2A> alpha2B2 and for (-)-noradrenaline alpha2B2 > alpha2Bl > alpha2A. All three alpha2-adrenoceptors showed the expected stereoselective binding for adrenaline enantiomers, the (+)-form being 7-10 fold less potent than the (-)form. 4. [3H]-yohimbine was also used as radioligand. The data with this ligand were fully compatible with the [3H]-RX821002 data. However, [3H]-yohimbine appeared to label only alpha2Bl- and alpha2B2-adrenoceptors presumably because it had too low an affinity for alpha2A-adrenoceptors. 5. We conclude that three pharmacological subtypes of alpha2-adrenoceptors are labelled by [3H]-RX821002 in the rat kidney. Guanoxabenz and ARC 239 may be used in competition studies to delineate between these three alpha2-adrenoceptor subtypes. Moreoever, we here present a method allowing the determination of binding constants for an arbitrary drug to the three alpha2-adrenoceptor subtypes.

Animals↗
PubMed1991

Rat spinal cord alpha 2-adrenoceptors are of the alpha 2A-subtype: comparison with alpha 2A- and alpha 2B-adrenoceptors in rat spleen, cerebral cortex and kidney using 3H-RX821002 ligand binding.

Binding of the alpha 2-adrenoceptor antagonist radioligand 3H-RX821002 was investigated in membranes from rat spinal cord, spleen, cerebral cortex and kidney. The ligand was found to bind to saturable binding sites with apparent uniform affinities within each tissue. Seven compounds, some of which have previously been reported to be selective for either alpha 2A- or alpha 2B-adrenoceptors, were used in competition with 3H-RX821002. By using computer modelling, competition curves generated for three of these compounds (ARC 239, prazosin and oxymetazoline) could be resolved into two site fits in the kidney, Kd's of the drugs being compatible with the notion that these sites corresponded to alpha 2A- and alpha 2B-adrenoceptors. Moreover, rauwolscine and yohimbine were found to be about 14 and 9-fold selective for alpha 2B-adrenoceptors in the kidney. In all other tissues studied drug competition curves were uniphasic and computer modelled into one site fits, drug Kd's being well correlated to those for the alpha 2A-adrenoceptor. In rat spinal cord 26 further drugs, which showed wide variation in structure, were evaluated in competition with 3H-RX821002. Of these compounds, competion curves of the agonists UK-14,304, (-) and (+) adrenaline were modelled into two site fits whereas those of the remaining compounds could be modelled only into one site fits. Since the high affinity site for UK-14,304, (-) and (+) adrenaline was eliminated when EDTA, Gpp(NH)p and 140 mM NaCl was present in the assay the heterogeniety observed in spinal cord was considered to be due to formation of high and low affinity conformations of the alpha 2-adrenoceptor for agonists. It is concluded that 3H-RX821002 is useful to label both alpha 2A- and alpha 2B-adrenoceptors in the rat. Moreover, the binding sites labelled by 3H-RX821002 in the spinal cord appear to consist of a single population of alpha 2-adrenoceptors of the alpha 2A-type.

Adrenergic alpha-Antagonists↗
PubMed1990

Antinociceptive actions of alpha 2-adrenoceptor agonists in the rat spinal cord: evidence for antinociceptive alpha 2-adrenoceptor subtypes and dissociation of antinociceptive alpha 2-adrenoceptors from cyclic AMP.

The antinociceptive actions of intrathecal injections of two alpha 2-adrenergic agonists, UK-14,304 and guanfacine, were investigated in rats after pretreatment of the animals with the noradrenaline neurotoxin N-2-chloroethyl-N-ethyl-2-bromobenzylamine (DSP4) 14 days in advance. The chronic noradrenaline depletion induced by DSP4 caused a marked increase in sensitivity of the antinociceptive action of UK-14,304 in the tail-flick test. By contrast, the antinociceptive effect of guanfacine was not appreciably affected by the DSP4 treatment. The antinociceptive effects of both UK-14,304 and guanfacine were blocked by intraperitoneal injections of yohimbine, a result indicating that both drugs induced their actions by activating alpha 2-adrenoceptors. Both UK-14,304 and guanfacine were found to reduce the production of cyclic AMP (cAMP) in the spinal cord, as determined using an in vitro radioisotopic method. The cAMP inhibitory effects of both agonists were effectively blocked by yohimbine, but not by prazosin, a finding indicating the alpha 2-adrenergic nature of the response. However, the cAMP inhibitory effect of UK-14,304 was not potentiated by pretreatment with DSP4, a finding in marked contrast with the strong potentiation of the antinociceptive action of UK-14,304 induced by the chronic depletion of endogenous noradrenaline. Moreover, intrathecal injections of forskolin, which increased the endogenous levels of spinal cord cAMP fivefold, did not modify the antinociceptive effects of UK-14,304 or guanfacine in neither normal nor DSP4-treated animals. It is suggested that there exist pharmacologically differing alpha 2-adrenergic receptor pathways capable of mediating antinociceptive effects at the level of the spinal cord. The cAMP inhibitory actions of spinal cord alpha 2-adrenoceptors appear not to be directly linked with the antinociceptive actions of these receptors.

Adrenergic alpha-Agonists↗
PubMed1990

Further characterization of the guinea pig cerebral cortex idazoxan receptor: solubilization, distinction from the imidazole site, and demonstration of cirazoline as an idazoxan receptor-selective drug.

We have demonstrated previously that [3H]idazoxan, besides being able to bind to alpha 2-adrenergic receptors, may also bind to a nonadrenergic idazoxan-receptor site with high affinity. The idazoxan receptor is tightly bound to cellular membranes, and we have now developed a method to solubilize it from the guinea pig cerebral cortex by using the detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). The CHAPS-solubilized receptor retains its binding properties for drugs: the membrane-bound, as well as the solubilized, idazoxan receptor shows high affinities for a number of imidazolines (cirazoline, idazoxan, tolazoline, naphazoline, tramazoline, clonidine, and oxymetazoline), some imidazoles (medetomidine, detomidine), and guanfacine. By contrast, catecholamines (adrenaline, noradrenaline, isoprenaline, and dopamine) and a number of other neurotransmitters and neuromodulators (serotonin, histamine, glutamic acid, gamma-aminobutyric acid, glycine, and adenosine) show negligible affinities for the idazoxan receptor. Moreover, the idazoxan receptor shows grossly different binding properties for histamine, cimetidine, and imidazole-4-acetic acid compared to what has been described for the nonadrenergic imidazole site labeled by p-[3H]amino-clonidine, indicating that the two receptor sites are distinct. Radioligand binding data further indicate that cirazoline is an idazoxan receptor-selective drug (KD = 1 nM) showing a 50-210-fold selectivity for binding to the idazoxan receptor when compared to alpha 2-adrenergic receptors and an about 500-fold selectivity when compared to alpha 1-adrenergic receptors. We have also reviewed the literature for possible nonadrenergic actions of idazoxan and cirazoline, and we suggest that idazoxan receptors might be involved in the control of prolactin release from the pituitary.

Adrenergic alpha-Agonists↗
PubMed1990

Functional supersensitivity of antinociceptive spinal cord alpha 2-adrenoceptors, induced by depletion of endogenous noradrenaline, is associated with an enhanced sensitivity for guanine nucleotide regulation of 3H-clonidine binding.

In spinal cords from normal mice, 3H-clonidine bound to alpha 2-adrenoceptors with an apparant Kd of 4.6 nM and capacity (Bmax) of 430 fmol/mg protein. The non-hydrolyzable GTP analogue Gpp(NH)p was found to dose-dependently reduce the binding of 3H-clonidine the effect of Gpp(NH)p being essentially maximal at 10(-4) M. Gpp(NH)p was found to reduce the apparent affinity as well as the apparent number of binding sites for 3H-clonidine; the Kd of 3H-clonidine being 16 nM and the Bmax 300 fmol/mg protein when 10(-4) M of the nucleotide was present. In mice pretreated with the noradrenaline neurotoxin DSP4 for 14 days prior to assay, the specific binding of nearly saturating concentrations (10-30 nM) of 3H-clonidine was virtually the same as for control mice. However, for the DSP4 treated animals the down-regulation of 3H-clonidine binding induced by 10(-4) M Gpp(NH)p was significantly higher than for control mice. Thus, at a 3H-clonidine concentration of 10 nM the down-regulation induced by Gpp(NH)p was 64.1 +/- 2.6% for the DSP4 versus 58.9 +/- 2.3% for the control mice (P less than 0.05). At a 3H-clonidine concentration of 30 nM the down-regulation was 52.4 +/- 1.4% for DSP4 versus 41.8 +/- 4.1% for control mice (P less than 0.05). DSP4 pretreatment also potentiated the antinociceptive effect of intrathecal clonidine, as assessed by the tail-flick and hot plate tests. It is suggested that the functional supersensitivity induced by DPS4 for clonidine is related to the increased sensitivity for guanine nucleotide regulation of the spinal cord alpha 2-adrenoceptors.

Animals↗
PubMed1989

Alpha 2-adrenoceptors mediate inhibition of cyclic AMP production in the spinal cord after stimulation of cyclic AMP with forskolin but not after stimulation with capsaicin or vasoactive intestinal peptide.

In slices obtained from the ventral and the dorsal guinea pig spinal cord both forskolin and vasoactive intestinal peptide (VIP) caused a dose-dependent stimulation of the production of cyclic AMP. By contrast capsaicin stimulated cyclic AMP formation only in the dorsal cord; no effect was observed in the ventral cord. The alpha 2-adrenergic agonist UK-14,304 dose-dependently inhibited the production of cyclic AMP in both the dorsal and ventral aspects of the cord when the formation of cyclic AMP had been stimulated with 3 microM forskolin, the maximal inhibition amounting to 25-32%. Also the basal (i.e., unstimulated) production of cyclic AMP was inhibited, the inhibition amounting to about 16-18%. However, after stimulation of cyclic AMP formation in the dorsal cord with capsaicin, UK-14,304 was virtually ineffective in inhibiting the accumulation of cyclic AMP. Also, when the formation of cyclic AMP was stimulated with VIP, UK-14,304 was virtually ineffective in inhibiting the formation of cyclic AMP both in the ventral and the dorsal parts of the cord. When cyclic AMP production had been stimulated with forskolin the ability of UK-14,304 to inhibit the formation of cyclic AMP was not attenuated by capsaicin, either in the ventral or in the dorsal cord. The results are discussed with the notion that cyclic AMP inhibitory spinal cord alpha 2-adrenoceptors are located on cells accessible to stimulation of cyclic AMP with forskolin but not with capsaicin or VIP.

Adenine↗
PubMed1989

Guanfacine inhibits excitatory amino acid release from rat cerebral cortex through a non-adrenergic mechanism.

The effect of guanfacine on the release of excitatory amino acids from rat cerebral cortex slices was assessed after preloading the slices with 3H-D-aspartate. It was found that guanfacine dose-dependently inhibited the veratridine stimulated release of the excitatory amino acid analogue; the ED50 of guanfacine being 6.4 +/- 2.3 microM (mean +/- S.E.M.) and the veratradine stimulated efflux being completely abolished at about 48 microM of guanfacine. Guanfacine also inhibited the veratridine stimulated release of endogenous aspartate and glutamate with similar potency to the inhibition of 3H-D-aspartate efflux. The inhibitory effect of guanfacine on the veratridine stimulated 3H-D-aspartate efflux was neither affected by high concentrations of the alpha 2-adrenoceptor blocker idazoxan nor by high concentrations of the alpha 1-adrenoceptor blocker prazosin. It is concluded that guanfacine inhibits the neural excitatory amino acid release in the rat cortex by a novel non-alpha 2/alpha 1-adrenergic mechanism.

Amino Acids↗