Visualization of neurokinin-3 receptor sites in rat brain using the highly selective ligand [3H]senktide

doi:10.1016/0006-8993(90)91218-6

Brain Research

Volume 506, Issue 1, 1 January 1990, Pages 175-179

https://doi.org/10.1016/0006-8993(90)91218-6 Get rights and content

Abstract

The autoradiographic distribution of the neurokinin (NK)-3 receptor sub-type was visualized in the rat brain using [³H]senktide, a highly selective ligand. [³H]Senktide apparently binds to a single class of high affinity (K_d = 2.8 ± 1.0nM), low capacity (B_max = 31.2 ± 3.0fmol/mg protein) sites in rat brain cortex. The ligand selectivity pattern reveals that eledoisin and senktide are potent competitors of both [³H]senktide and [¹²⁵I]Bolton-Hunter eledoisin binding sites demonstrating the NK-3 nature of these sites. Autoradiographic data show that [³H]senktide binding sites are concentrated in mid-cortical layers, supraoptic nucleus, zona incerta, basolateral nucleus of the amygdala and interpeduncular nucleus. Much lower densities of binding are seen in most other areas such as the caudate-putamen and cerebellum. This distribution is similar, but not identical, to that previously reported for NK-3 sites using less selective ligands. It is most likely because less selective probes also bind to other classes of NK receptors. The higher selectivity of [³H]senktide is thus an important advantage for the precise characterization of NK-3 receptor binding parameters.

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Cited by (96)

Neurokinin receptors in drug and alcohol addiction
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Citation Excerpt :
This is intriguing because the NK3R is expressed more highly than the NK1R in the rodent VTA; however, retrograde tracing studies show that the NK1R seems to be expressed more strongly in VTA neurons projecting to the NAC, whereas NK3R is more prominent in VTA neurons that project to the frontal cortex (Lessard, 2009). Like the NK1R, the NK3R is expressed widely throughout the brain in regions that mediate affective and motivated behaviors, including in the olfactory bulb, amygdala, multiple cortical regions, hypothalamic subnuclei, hippocampus, locus coeruleus, VTA, and interpenduncular nucleus, among others (Massi et al., 2000; Stoessl and Hill, 1990; Dam et al., 1990; Shughrue et al., 1996). Overall, NK3R activation has very similar effects to NK1R stimulation on mesolimbic DA function.
The neurokinins are a class of peptide signaling molecules that mediate a range of central and peripheral functions including pain processing, gastrointestinal function, stress responses, and anxiety. Recent data have linked these neuropeptides with drug-related behaviors. Specifically, substance P (SP) and neurokinin B (NKB), have been shown to influence responses to alcohol, cocaine, and/or opiate drugs. SP and NKB preferentially bind to the neurokinin-1 receptor (NK1R) and neurokinin-3 receptor (NK3R), respectively, but do have some affinity for all classes of neurokinin receptor at high concentrations. NK1R activity has been shown to influence reward and reinforcement for opiate drugs, stimulatory and neurochemical responses to cocaine, and escalated and stress-induced alcohol seeking. In reinstatement models of relapse-like behavior, NK1R antagonism attenuates stress-induced reinstatement for all classes of drugs tested to date. The NK3R also influences alcohol intake and behavioral/neurochemical responses to cocaine, but less research has been performed in regard to this particular receptor in preclinical models of addiction. Clinically, agents targeting these receptors have shown some promise, but have produced mixed results. Here, the preclinical findings for the NK1R and NK3R are reviewed, and discussion is provided to interpret clinical findings. Additionally, important factors to consider in regards to future clinical work are suggested.
The neurokinin-3 receptor agonist senktide facilitates the integration of memories for object, place and temporal order into episodic memory
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Senktide, a potent neurokinin-3 receptor (NK3-R) agonist, has been shown to have promnestic effects in adult and aged rodents and to facilitate episodic-like memory (ELM) in mice when administrated before the learning trial. In the present study we assessed the effects of senktide on memory consolidation by administering it post-trial (after the learning trial) in adult rats. We applied an ELM test, based on the integrated memory for object, place and temporal order, which we developed (Kart-Teke, de Souza Silva, Huston, & Dere, 2006). This test involves two learning trials and one test trial. We examined intervals of 1 h and 23 h between the learning and test trials (experiment 1) in untreated animals and found that they exhibited intact ELM after a delay of 1 h, but not 23 h. In another test for ELM performed 7 days later, vehicle or senktide (0.2 mg/kg, s.c.) was applied immediately after the second learning trial and the test was conducted 23 h later (experiment 2). Senktide treatment recovered components of ELM (memory for place and object) compared with vehicle-treated animals. After one more week, vehicle or senktide (0.2 mg/kg, s.c.) was applied post-trial and the test conducted 6 h later (experiment 3). The senktide-treated group exhibited intact ELM, unlike the vehicle-treated group. Finally, animals received post-trial treatment with either vehicle or SR142801, a selective NK3-R antagonist (6 mg/kg, i.p.), 1 min before senktide injection (0.2 mg/kg, s.c.) in the ELM paradigm and were tested 6 h later (experiment 4). The vehicle + senktide group showed intact ELM, while the SR142801 + senktide group did not. The results indicate that senktide facilitated the consolidation or the expression of ELM and that the senktide effect was NK3-R dependent.
Is there still a future for neurokinin 3 receptor antagonists as potential drugs for the treatment of psychiatric diseases?
2012, Pharmacology and Therapeutics
Citation Excerpt :
The distribution of NK3 binding sites in the CNS has been studied in several species including rats, guinea pigs, gerbils and humans, by various techniques: radioligand autoradiography, in situ hybridization and immunohistochemistry. These studies revealed important species differences (Almeida et al., 2004; Buck et al., 1986; Carpentier & Baude, 1996; Dam et al., 1990; Ding et al., 1996; Langlois et al., 2001; Mussap & Burcher, 1990; Pinto et al., 2004; Saffroy et al., 2003; Shughrue et al., 1996; Stoessl & Hill, 1990; Yip & Chahl, 2001). While in rats, guinea pigs and gerbils the NK3 receptor was similarly distributed within the cerebral cortex, the zona incerta, the medial habenula, the amygdala nuclei, the superior colliculus, the interpeduncular nucleus, the ventral tegmental area, the substantia nigra pars compacta and the dentate gyrus, outside of these structures each species displayed a specific distribution pattern of central NK3 receptors (Fig. 1).
Selective non-peptide antagonists for the neurokinin 3 (NK₃) receptor first became available about twenty years ago. Although the understanding of the role of the NK₃ receptor in the brain has been greatly complicated by marked species differences in its distribution and by pharmacological heterogeneity, studies with brain-penetrant non-peptide NK₃ receptor antagonists in animals have indicated that these compounds may find utility in a number of psychiatric diseases, including schizophrenia, anxiety and depressive disorders. However, clinical studies with selective NK₃ receptor antagonists in these psychiatric conditions have been disappointing and they were unable to confirm the promising therapeutic potential from animal studies, thereby questioning the therapeutic utility of these compounds for CNS disorders. The purpose of this article is to provide a critical overview of the available data on NK₃ receptor antagonists in the psychiatry research and development field, by reviewing the behavioral and neurochemical effects of these agents in both preclinical and clinical studies.
Neurokinin3-R agonism in aged rats has anxiolytic-, antidepressant-, and promnestic-like effects and stimulates ACh release in frontal cortex, amygdala and hippocampus
2011, European Neuropsychopharmacology
Citation Excerpt :
The effect of senktide on immobility could be mediated through an interaction with noradrenaline, serotonin, and/or other neurotransmitters in the brain classically involved in depression. For example, in the raphe nuclei NK3 binding sites have been shown (Dam et al., 1990), and these were decreased in the 5,7-dihydroxytryptamine-treated median raphe nucleus (Stoessl and Hill, 1990), thus, providing a possibility of neurokinin and serotonergic interactions. Moreover, senktide has been shown to activate local glutamatergic inputs to serotonergic neurons of the dorsal raphe nucleus (Liu et al., 2002).
Neurokinin-3 receptors (NK₃-R) are localized in brain regions which have been implicated in processes governing learning and memory as well as emotionality. The effects of acute subcutaneous (s.c.) senktide (0.2 and 0.4 mg/kg), a NK₃-R agonist, were tested in aged (23–25 month old) Wistar rats: (a) in an episodic-like memory test, using an object discrimination task (this is the first study to test for deficits in episodic-like memory in aged rats, since appropriate tests have only recently became available); (b) on parameters of anxiety in an open field test, (c) on indices of depression in the forced swimming test and (d) on the activity of cholinergic neurons of the basal forebrain, using in vivo microdialysis and HPLC. Neither the saline-, nor senktide-treated aged animals, exhibited episodic-like memory. However, the senktide-, but not the vehicle-treated group, exhibited object memory for spatial displacement, a component of episodic memory. Senktide injection also had anxiolytic- and antidepressant-like effects. Furthermore, the active doses of senktide on behavior increased ACh levels in the frontal cortex, amygdala and hippocampus, suggesting a relationship between its cholinergic and behavioral actions. The results indicate cholinergic modulation by the NK₃-R in conjunction with a role in the processing of memory and emotional responses in the aged rat.
In vitro and in vivo comparison of two non-peptide tachykinin NK<inf>3</inf> receptor antagonists: Improvements in efficacy achieved through enhanced brain penetration or altered pharmacological characteristics
2010, European Journal of Pharmacology
Clinical evaluation of tachykinin NK₃ receptor antagonists has provided support for the therapeutic utility of this target in schizophrenia. However, these studies have not been entirely conclusive, possibly because of the pharmacokinetic limitations of these molecules. In the search for tachykinin NK₃ receptor antagonists with improved properties, we have discovered GSK172981 and GSK256471. Both compounds demonstrated high affinity for recombinant human (pK_i values 7.7 and 8.9, respectively) and native guinea pig (pK_i values 7.8 and 8.4, respectively) tachykinin NK₃ receptors. In vitro functional evaluations revealed GSK172981 to be a competitive antagonist (pA₂ = 7.2) at cloned human tachykinin NK₃ receptor whereas GSK256471 diminished the neurokinin B-induced E_max response, indicative of non-surmountable antagonist pharmacology (pA₂ = 9.2). GSK172981 also exhibited a competitive profile in antagonizing neurokinin B-stimulated neuronal activity recorded from the guinea pig medial habenula slices (apparent pK_B = 8.1), whilst GSK256471 abolished the agonist-induced response. Central nervous system penetration by GSK172981 and GSK256471 was indicated by dose-dependent ex vivo tachykinin NK₃ receptor occupancy in medial prefrontal cortex (ED₅₀ values of 0.8 and 0.9 mg/kg, i.p., respectively) and the dose-dependent attenuation of agonist-induced “wet dog shake” behaviours in guinea pigs. Finally, in vivo microdialysis studies demonstrated that acute GSK172981 (30 mg/kg, i.p.) and GSK256471 (1 mg/kg, i.p.) attenuated haloperidol-induced increases in extracellular dopamine in the guinea pig nucleus accumbens. Taken together, these in vitro and in vivo characterisations of the tachykinin NK₃ receptor antagonists GSK172981 and GSK256471 support their potential utility in the treatment of schizophrenia.
Blockade of neurokinin-3 receptors modulates dopamine-mediated behavioral hyperactivity
2009, Neuropharmacology
Citation Excerpt :
NK-3 receptors are found peripherally on nerve endings of primary afferent neurons innervating the respiratory, gastrointestinal and urinary tracts (Patacchini and Maggi, 2001), however they are also differentially expressed in the central nervous system. NK-3 receptors localized in the substantia nigra, ventral tegmental area (VTA) and prefrontal cortex (Dam et al., 1990) are thought to regulate dopaminergic neurotransmission and locomotive behaviors (Elliott et al., 1991; Overton et al., 1992). Tyrosine hydroxylase-containing neurons in the substantia nigra and VTA have been shown to express NK-3 receptors (Chen et al., 1998).
Acute activation or blockade of neurokinin-3 (NK-3) receptors has been shown to alter dopamine-mediated function and behaviors, however long-term effects of NK-3 receptor blockade remain largely unknown. The present study investigated whether acute and repeated administration of the NK-3 receptor antagonist SB 222200 altered hyperactivity induced by cocaine, and examined its effects on dopamine D1 receptor density in the striatum. Adult male CD-1 mice received either vehicle or SB 222200 (2.5 or 5 mg/kg, s.c.) 30 min before a cocaine injection (20 mg/kg, i.p.) and behavioral responses were recorded. Mice that were administered SB 222200 had an attenuated stereotypic response to cocaine compared to vehicle treated mice. Mice were also injected once daily with either vehicle or SB 222200 (5 mg/kg, s.c.) for 5 days, and after a 7-day drug-free period they were challenged with either saline, cocaine or the dopamine D1 receptor agonist SKF 82958 (0.125 or 0.25 mg/kg, i.p.). Mice injected with SB 222200 had significantly enhanced hyperactivity when challenged with cocaine or a low dose of SKF 82958 (0.125 mg/kg, i.p.) compared to control mice. Brains of mice administered vehicle or SB 222200 for 5 days were harvested after a 7-day drug-free period for dopamine D1 receptor quantification by radioligand binding. [³H] SCH 23390 homogenate binding studies showed a 19.7% increase in dopamine D1 receptor density in the striatum of SB 222200 treated mice. These data suggest that repeated blockade of NK-3 receptors enhances subsequent dopamine-mediated behaviors possibly resulting from dopamine D1 receptor up-regulation in the striatum.

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Visualization of neurokinin-3 receptor sites in rat brain using the highly selective ligand [3H]senktide

Abstract

Neuroscience

Trends Pharmacol. Sci.

Life Sci.

Trens Neurosci.

J. Biol. Chem.

Eur. J. Pharmacol.

J. Biol. Chem.

Trends Neurosci.

Regul. Peptides

A new type of tachykinin binding site in the rat brain characterized by the specific binding of a labelled elodoisin derivative

Mol. Pharmacol.

Visualization of neurokinin-3 receptor sites in rat brain using the highly selective ligand [³H]senktide