Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P08908 (5-HT1A)
 5,574 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

In this study, [125I]iodocyanopindolol ([125I]ICYP), in the presence of isoproterenol, was used to label 5-hydroxytryptamine1B (5-HT1B) receptors in homogenates of the cortex, substantia nigra and caudate-putamen of the rat. The determination of the appropriate concentrations of isoproterenol required to block optimally beta adrenoceptors whereas producing minimal occupancy of 5-HT1B receptors was achieved by generating isotherms for isoproterenol at multiple concentrations of [125I]ICYP. When different concentrations of isoproterenol were used with increasing concentrations of [125I]ICYP, a linear Scatchard transformation of the saturation curve was achieved, even with ligand concentrations about 6-fold greater than the KD for [125I]ICYP. Competition for [125I]ICYP (100 pM) labeled binding sites by 15 serotonin agonists or antagonists was adequately described by a single site model, and the affinity of these drugs for the site labeled by [125I]ICYP was similar to that determined previously when using indirect methods to label 5-HT1B receptors. Serotonin itself showed high affinity for this binding site as did two antagonists, metergoline and methiothepin. By contrast, drugs thought to be selective for the 5-HT1A receptor (e.g., 8-hydroxy-2-(di-n-propylamino)tetralin, buspirone and spiperone) showed very weak affinity for the binding site labeled with [125I]ICYP. The effect of nucleotide regulation on [125I]ICYP binding at 5-HT1B receptors also was evaluated. It was determined that GTP had little effect on the binding of [125I]ICYP, reducing total binding by only 15% and shifting the displacement curve of 5-HT by a factor of less than two. The regulation of 5-HT1B receptors, labeled by [125I]ICYP, also was evaluated. Intraventricular injections of 5.7-dihydroxytryptamine increased significantly the number of 5-HT1B receptors in the caudate-putamen; this treatment had no effect on 5-HT1B binding sites either in the cortex or substantia nigra. The regulatable binding site for [125I]ICYP in the caudate-putamen had a pharmacological profile very similar to that of the 5-HT1B binding site in the cortex. [125I]ICYP appears to be a useful ligand to measure 5-HT1B receptors in the brain of the rat. The localized increase in 5-HT1B receptors in the caudate-putamen after destruction of central serotonergic neurons might indicate that the majority of 5-HT1B receptors in this area of brain are not located on serotonergic nerve terminals.
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PMID:Application of [125I]iodocyanopindolol to measure 5-hydroxytryptamine1B receptors in the brain of the rat. 333 96

In vitro investigations revealed that PAT (8-hydroxy-2-(n-dipropylamino)tetralin) interacted with postsynaptic 5-HT receptors in the rat brain: the drug stimulated 5-HT-sensitive adenylate cyclase in homogenates of colliculi from new-born rats (KAapp 8.6 microM) and inhibited the specific binding of [3H]5-HT to 5-HT1 sites. The PAT-induced inhibition of [3H]5-HT binding showed marked regional differences compatible with a preferential interaction of PAT (IC50 2 nM) with the 5-HT1A subclass. As previously seen with 5-HT agonists, the efficacy of PAT for displacing [3H]5-HT bound to hippocampal membranes was markedly increased by Mn2+ (1 mM) and reduced by GTP (0.1 mM). PAT also affected presynaptic 5-HT metabolism since it inhibited competitively (Ki 1.4 microM) [3H]5-HT uptake into cortical synaptosomes and reduced (in the presence of the 5-HT uptake inhibitor fluoxetine) the K+-evoked release of [3H]5-HT previously taken up or newly synthesized from [3H]tryptophan in cortical or striatal slices. This latter effect was prevented by 5-HT antagonists (methiothepin, metergoline) suggesting that it was mediated by the stimulation of presynaptic 5-HT autoreceptors by PAT. Like 5-HT, PAT counteracted the stimulatory effect of K+-induced depolarization on the synthesis of [3H]5-HT from [3H]tryptophan in cortical slices. It is concluded that PAT is a potent 5-HT agonist acting on both post- and presynaptic 5-HT receptors in the rat brain.
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PMID:Biochemical evidence for the 5-HT agonist properties of PAT (8-hydroxy-2-(di-n-propylamino)tetralin) in the rat brain. 620 61

We have investigated the distribution of [3H]5-carboxamidotryptamine ([3H]5-CT) binding sites by in vitro autoradiography on sections of guinea-pig and rat brain. In saturation studies, the ligand recognised a saturable, homogeneous population of binding sites with an affinity ranging from 0.19-0.45 nM depending on the region. The labelling pattern was heterogeneous, and the displacement pattern with different competing drugs selective for different 5-HT receptor subtypes was complex. [3H]5-CT appeared to label 5-HT1B/5-HT1D sites in the substantia nigra, globus pallidus and caudate/putamen, as the binding in these regions was displaced by the 5-HT1B/1D receptor selective agents sumatriptan, CP-122,288 and GR-127,935. In the hippocampus and lateral septum, the very dense [3H]5-CT binding was displaced with high affinity by the 5-HT1A receptor selective agonist 8-hydroxy-dipropylaminotetralin ((+/-)-8-OH-DPAT), dihydroergotamine and 5-HT. In contrast the affinity of the 5-HT1 receptor antagonists spiperone and methiothepine was much lower than their previously published potency at 5-HT1A receptors. The affinity of agonists, taken together with the fact that the distribution of these [3H]5-CT sites overlaps that of [3H]8-OH-DPAT binding sites in serial sections, suggest that these sites correspond to 5-HT1A receptors. Their atypical properties deserve further investigations. While [3H]5-CT binding at 5-HT1B/1D sites and these atypical 5-HT1A sites was inhibited by the GTP analogue 5'-beta, gamma-imidotriphosphate, [3H]5-CT binding in the superficial cortical layers and in midline thalamic nuclei was insensitive to this agent. It was however displaced by low concentrations of spiperone, clozapine and methiothepine, but not by sumatriptan, CP-122,288, GR-127,935 or dihydroergotamine. This binding profile is similar to that of 5-HT7 receptors, while the spatial distribution of these sites matches the known distribution of 5-HT7 messenger RNA. We did not find evidence of [3H]5-CT labelling to 5-HT5 receptors, in spite of their reported high affinity for this ligand. It is concluded that [3H]5-CT, in the presence of selective blockers, can be used to investigate the properties of 5-HT1A, 5-HT1B/1D and 5-HT7 receptors in the rodent brain, although further studies are required to explain the atypical features of [3H]5-CT binding in 5-HT1A receptors containing regions.
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PMID:Autoradiographic visualisation of [3H]5-carboxamidotryptamine binding sites in the guinea pig and rat brain. 749 19

The human 5-HT1A receptor was expressed in Sf9 insect cells to examine desensitization as manifested by agonist-induced uncoupling from G proteins and second messengers. New binding sites were detected after infection of cells with the 5-HT1A receptor-bearing baculovirus. 5-HT1A receptor agonists caused inhibition of cAMP accumulation that could be attenuated by specific receptor antagonists. Brief pretreatment with 5-HT resulted in (1) an uncoupling of receptor from G proteins as evidenced by a loss of high-affinity agonist binding sites and a diminished ability of the receptor to increase incorporation of AA-GTP into endogenous Go alpha-like G proteins, (2) a decreased ability of the receptor to inhibit cAMP accumulation, and (3) increased phosphorylation of the 5-HT1A receptor on serine and threonine residues. Phosphorylation occurred in the presence of a number of cyclic nucleotide dependent kinase inhibitors, and desensitization of the cAMP response occurred in the presence of H-7 and also in cells with prolonged exposure to PMA. Both phosphorylation and desensitization were markedly attenuated by 100 nM and 1 microM heparin and demonstrated similar time courses and concentration-response relationships. Those results demonstrate a close association between agonist-induced desensitization and phosphorylation of the 5-HT1A receptor in Sf9 cells through a pathway that mainly does not involve protein kinase A or C and might involve a G protein-linked receptor kinase.
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PMID:Agonist-induced desensitization and phosphorylation of human 5-HT1A receptor expressed in Sf9 insect cells. 754 32

1. We investigated the effects of serotonin (5-hydroxytryptamine, 5-HT) on whole-cell barium currents through calcium channels in visualized neonatal rat hypoglossal motoneurones (HMs) in a thin brainstem slice preparation. 2. High voltage-activated (HVA) currents were elicited by depolarizing voltage steps from -70 to 0 mV; low voltage-activated (LVA) currents were evoked using steps to between -30 and -40 mV from hyperpolarized potentials (< -80 mV). 5-HT (1.0 microM) inhibited HVA currents by at least 10% in 70% of HMs tested (n = 99); in those responsive neurones, 5-HT decreased HVA current by 22 +/- 1.3% (mean +/- S.E.M.). In contrast, 5-HT had no effect on LVA current amplitude in HMs (n = 7). 3. Calcium current inhibition was mimicked by 5-carboxamidotryptamine maleate (5-CT), a 5-HT1 receptor agonist, and by R(+)-8-hydroxydipropylaminotetralin hydrobromide (8-OH-DPAT), a specific 5-HT1A agonist; N-(3-trifluoromethylphenyl) piperazine hydrochloride (TFMPP), a 5-HT1B agonist, was without effect. The effect of 5-HT was blocked by the 5-HT1A antagonist 1-(2-methoxyphenyl)-4-[4-(2-phthalimido)butyl]piperazine hydrobromide (NAN-190) but not by ketanserin, a 5-HT2A/2C antagonist. Although R(-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane hydrochloride (DOI), a 5-HT2A/2C agonist, mimicked the current inhibition by 5-HT, it was ineffective in the presence of NAN-190. These data indicate that 5-HT1A receptors mediate calcium current inhibition by 5-HT. 4. Following application of either omega-conotoxin-GVIA (omega-CgTX) or omega-agatoxin-IVA (omega-Aga-IVA), to block N- and P-type components of calcium current, the 5-HT-sensitive current was reduced; 5-HT had no effect on the current remaining after application of both toxins. Thus, 5-HT inhibits both N- and P-type calcium currents in neonatal HMs. 5. Inhibition of HVA current by 5-HT was irreversible, and subsequent applications of 5-HT were occluded, when GTP gamma S was substituted for GTP in the pipette. In addition, inhibition of HVA current by 5-HT was relieved following depolarizing prepulses. These data indicate that inhibition of calcium channels by 5-HT is mediated by G proteins. 6. Under current clamp, both 5-HT and 8-OH-DPAT decreased the amplitude of the after-hyperpolarization (AHP) that followed action potentials, indicating involvement of a 5-HT1A receptor.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:Inhibition of N- and P-type calcium currents and the after-hyperpolarization in rat motoneurones by serotonin. 756 6

The high-affinity GTP hydrolyzing activity stimulated by 5-hydroxytryptamine (5-HT) receptor agonists was pharmacologically characterized in rat hippocampal membranes. The addition of 100 microM 5-HT increased significantly the Vmax of high-affinity GTPase activity with an apparent Km of 0.37 microM in a Mg(++)-dependent fashion. 5-HT receptor agonists, except for the selective 5-HT2 receptor agonists, (+/-)-1-(2,5-dimethoxy-4-bromophenyl)-2-aminopropane and (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane, stimulated the activity in a concentration-dependent manner, with affinities indicative of the 5-HT1A receptor involvement. 2-(2,6-Dimethoxyphenoxyethyl)aminomethyl-1,4-benzodioxane, buspirone, ipsapirone, metergoline and (+/-)-8-hydroxy-2-(di-n-propylamino)tetralin showed activities characterizing these as partial agonists. The drug 1-(2-methoxyphenyl)-4-[4-(2-phthalimido)butyl]piperazine was also characterized as a weak partial agonist. 5-HT (100 nM)-stimulated activity was potently antagonized by metitepine (also called methiothepin) and spiperone (with a Kb value of 37 nM in a competitive manner) but not by ketanserin. The affinities of the agonists obtained in this study correlated well with those for the 5-HT1A receptor-mediated inhibition of forskolin-stimulated adenylyl cyclase activity in guinea pig and rat hippocampal membranes reported in a previous article. The 5-HT-mediated activation of high-affinity GTPase in rat hippocampal membranes can be used to investigate a functional interaction between the 5-HT1A receptors and G proteins, in particular the Gi subfamily, associated with adenylyl cyclase inhibition.
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PMID:Pharmacological characterization of the 5-hydroxytryptamine-1A receptor-mediated activation of high-affinity GTP hydrolysis in rat hippocampal membranes. 761 17

This study addresses the mechanisms responsible for regulation of high-conductance anion channels by GTP binding proteins in Chinese hamster ovary (CHO) cells. Single-channel currents were measured in inside-out membrane patches using patch-clamp techniques. Anion-selective channels with a unitary conductance of 381 +/- 8 pS activated spontaneously in 48% of excised patches. In patches with no spontaneous channel activity, addition of GppNHp, a nonhydrolyzable analogue of GTP, activated channels in 8 of 12 studies, and in patches with spontaneous channel activity, GppNHp increased open probability in 4 of 4 experiments. In contrast, GDP beta S, a nonhydrolyzable GDP analogue, inhibited both spontaneous and GppNHp-induced channel activity. In patches without spontaneous channel activity, addition of cholera toxin activated channels in five of eight studies. Interestingly, pertussis toxin had a similar effect, activating channels in five of seven previously quiescent patches. To further evaluate the possible role of inhibitory G proteins in channel regulation, activity was measured in cell-attached patches in cells transfected with the serotonin 5-HT1A receptor, which is coupled to effector mechanisms through a pertussis toxin-sensitive G protein. Stimulation of 5-HT1A-transfected cells with the receptor agonist (+/-)-8-hydroxy-2-(di-n-propylamino)tetralin caused a transient decrease in open probability in either standard or high-potassium solutions. In aggregate, these findings suggest that both cholera and pertussis toxin-sensitive G proteins contribute to regulation of high-conductance anion channels in CHO cells.
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PMID:Regulation of high-conductance anion channels by G proteins and 5-HT1A receptors in CHO cells. 768 Dec 62

Binding characteristics of a novel radioiodinated ligand, [125I]R(+)trans-7-hydroxy-2-(N-n-propyl-N-3'-iodo-2'-propenyl) aminotetralin ([125I]R(+)trans-7-OH-PIPAT), were evaluated using homogenate binding and autoradiographic techniques in rat brain. [125I]R(+)trans-7-OH-PIPAT bound to sites (dopamine receptors) in homogenates of rat basal forebrain (including caudate putamen, nucleus accumbens and olfactory tubercle) with a high affinity (Kd = 0.42 nM). A majority (70%) of the sites labeled by [125I]R(+)trans-7-OH-PIPAT in basal forebrain were GTP-sensitive. In rat hippocampal homogenates, specific and saturable binding of [125I]R(+)trans-7-OH-PIPAT to 5-HT1A receptors, with a Kd value of 1.4 nM and a Bmax value of 210 fmol/mg protein, was observed. Binding of [125I]R(+)trans-7-OH-PIPAT to sigma sites was also demonstrated in rat cerebellar homogenates. In the presence of GTP (to inhibit binding to D2 and 5-HT1A receptors) and DTG (to inhibit binding to sigma sites), dopamine D3 receptors could be selectively labeled with [125I]R(+)trans-7-OH-PIPAT. [125I]R(+)trans-7-OH-PIPAT offers several unique advantages, including high specific activity and high affinity binding, which make it an excellent probe for the investigation and characterization of the distribution of dopamine D3 receptors.
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PMID:Characterization of binding sites for [125I]R(+)trans-7-OH-PIPAT in rat brain. 770 18

The tritiated derivative of the novel silent 5-HT1A receptor antagonist WAY 100635 [N-(2-(4-(2-methoxyphenyl)-1-piperazinyl)ethyl)-N-(2-pyridinyl) cyclohexane carboxamide] was tested as a potential radioligand of 5-HT1A receptors in the rat brain. Binding assays with membranes from various brain regions showed that [3H]WAY 100635 specifically bound to a homogeneous population of sites, with a Kd of 0.10 nM. The regional distribution of [3H]WAY 100635 specific binding sites, as assessed in membrane binding assays and by autoradiography of labelled brain sections, superimposed exactly over that of 5-HT1A receptors specifically labelled by [3H]8-hydroxy-2-(di-n-propylamino) tetralin ([3H]8-OH-DPAT). Furthermore, the positive correlation (r = 0.96) between the respective pKi values of a large series of ligands as inhibitors of the specific binding of [3H]WAY 100635 and [3H]8-OH-DPAT in hippocampal membranes indicated that their pharmacological properties were similar. Nevertheless, marked differences also existed between [3H]8-OH-DPAT and [3H]WAY 100635 specific binding, as the former was inhibited by 1-100 microM GTP and GppNHp, whereas the latter was enhanced by these guanine nucleotides. In contrast, Mn2+ (1-10 mM) increased the specific binding of [3H]8-OH-DPAT, but inhibited that of [3H]WAY 100635. Treatment of membranes with N-ethylmaleimide (1-5 mM) markedly reduced their capacity to specifically bind [3H]8-OH-DPAT, but slightly increased (at 1 mM) or did not affect (at 5 mM) their [3H]WAY 100635 specific binding capacity. Finally, the Bmax of [3H]WAY 100635 specific binding sites was regularly 50-60% higher than that of [3H]8-OH-DPAT in the same membrane preparations from various brain regions (hippocampus, septum, cerebral cortex). These data are compatible with the idea that whereas [3H]8-OH-DPAT only binds to G-protein-coupled 5-HT1A receptors, [3H]WAY 100635 is a high affinity ligand of both G-protein-coupled and free 5-HT1A receptor binding subunits in brain membranes.
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PMID:The selective 5-HT1A antagonist radioligand [3H]WAY 100635 labels both G-protein-coupled and free 5-HT1A receptors in rat brain membranes. 772 Jul 79

GTP hydrolyzing activity was assayed by measuring the amount of 32P(i) released from 0.3 microM [gamma-32P]GTP in the membranes prepared from rat brain. 5-Hydroxytryptamine (5-HT) stimulated the high-affinity GTPase activity in hippocampus, but not in striatum, in a concentration-dependent manner with an EC50 value of 18 nM and maximal percent stimulation of 13.9%. This response was mimicked by (+/-)-8-hydroxy-2-(di-n-propylamino)tetralin [(+/-)-8-OH-DPAT], but not by (+/-)-1-(2,5-dimethoxy-4-iodophyenyl)-2-aminopropane [(+/-)-DOI]. These results suggest that 5-HT-stimulated high-affinity GTPase activity of the GTP-binding protein(s) is mediated via 5-HT1A receptor subtype in the rat hippocampus.
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PMID:5-HT1A receptor-mediated activation of high-affinity GTPase in rat hippocampal membranes. 777 84


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