Activation Of Neurokinin-3 Receptors Research Articles

NK3R signalling in the posterodorsal medial amygdala is involved in stress-induced suppression of pulsatile LH secretion in female mice.

Psychosocial stress negatively impacts reproductive function by inhibiting pulsatile luteinizing hormone (LH) secretion. The posterodorsal medial amygdala (MePD) is responsible in part for processing stress and modulating the reproductive axis. Activation of the neurokinin 3 receptor (NK3R) suppresses the gonadotropin-releasing hormone (GnRH) pulse generator, under hypoestrogenic conditions, and NK3R activity in the amygdala has been documented to play a role in stress and anxiety. We investigate whether NK3R activation in the MePD is involved in mediating the inhibitory effect of psychosocial stress on LH pulsatility in ovariectomised female mice. First, we administered senktide, an NK3R agonist, into the MePD and monitored the effect on pulsatile LH secretion. We then delivered SB222200, a selective NK3R antagonist, intra-MePD in the presence of predator odour, 2,4,5-trimethylthiazole (TMT) and examined the effect on LH pulses. Senktide administration into the MePD dose-dependently suppresses pulsatile LH secretion. Moreover, NK3R signalling in the MePD mediates TMT-induced suppression of the GnRH pulse generator, which we verified using a mathematical model. The model verifies our experimental findings: (i) predator odour exposure inhibits LH pulses, (ii) activation of NK3R in the MePD inhibits LH pulses and (iii) NK3R antagonism in the MePD blocks stressor-induced inhibition of LH pulse frequency in the absence of ovarian steroids. These results demonstrate for the first time that NK3R neurons in the MePD mediate psychosocial stress-induced suppression of the GnRH pulse generator.

Structural insights into neurokinin 3 receptor activation by endogenous and analogue peptide agonists

Neurokinin 3 receptor (NK3R) is a tachykinin receptor essential for the hypothalamic-pituitary-gonadal axis. The endogenous peptide agonist neurokinin B (NKB) preferentially activates NK3R, while substance P (SP) binds preferentially to NK1R. In addition, the SP analogue senktide more potently activates NK3R than NKB and SP. However, the mechanisms of preferential binding of peptide and NK3R activation remain elusive. Herein, we determined the cryogenic electron microscopy (cryo-EM) structures of the NK3R–Gq complex bound to NKB, SP and senktide. The three NK3R–Gq/peptide complexes utilize a class of noncanonical receptor activation mechanisms. Combining the structural analysis and functional assay illustrated that the consensus C-termini of the three peptide agonists share a conserved binding mode to NK3R, while the divergent N-termini of the peptides confer the preferential binding of the agonist to NK3R. In addition, the specific interactions between the N-terminus of senktide and the N-terminus and extracellular loops (ECL2 and ECL3) of NK3R lead to the improved activation displayed by senktide compared to SP and NKB. These findings pave the way to understand tachykinin receptor subtype selectivity and provide ideas to rationally develop drugs targeting NK3R.

How does neurokinin 3 receptor agonism affect pathological and cognitive impairments in an Alzheimer's disease-like rat model?

Alzheimer's disease (AD) is accepted as a form of progressive dementia. Cholinergic systems are commonly affected in AD. Neurokinin 3 receptor (NK3R) is involved in learning memory-related processes. It is known that the activation of NK3R affects the release of many neurotransmitters. The aim of this project was to investigate the effects of NK3R agonist senktide administration on neurobehavioral mechanisms in the experimental AD-like rat model. 50 male Wistar albino rats were divided into Control (C), AD, Control + NK3R agonist (CS), AD + NK3R agonist (ADS), AD + NK3Ragonist + antagonist groups (ADSO). We designed AD-like model by intrahippocampal administration of Aβ1-42. After NK3R agonist + antagonist injections, open field (OF), Morris water maze (MWM) tests were applied. Cholinergic mechanism analysis from hippocampus-cortex tissues was performed by ELISA and catecholamine analysis from brain stem tissue were performed by HPLC method. The transitions from edge to center, rearing, grooming parameters were found to be reduced in final values of OF. While the group-time interaction was significant in the OF test findings, there was no significant difference between the groups. In MWM test, ADS group showed a learning level close to control group and animals in AD and ADSO groups could not learn target quadrant in MWM test. The brain stem NA and DA concentrations were not statistically significant. Hippocampal AChE-ChAT levels were supported by positive effects of senktide on learning via the cholinergic mechanisms. As a result, NK3R agonists were found to be effective in improving cognitive functions in rats with AD pathology. In the experimental AD model, positive effects of NK3R on learning memory may be mediated by cholinergic mechanisms.

SAT-425 NKB Signaling in the Posterodorsal Medial Amygdala Stimulates Gonadotropin Release in a Kisspeptin-Independent Manner in Female Mice

Female reproduction is regulated by a complex signaling network, the function of which is dependent on the level of circulating estrogens. Neurokinin B (NKB) colocalizes with kisspeptin in Kiss1 neurons of the arcuate nucleus (ARC). The autosynaptic action of NKB via the neurokinin 3 receptor (NK3R) in these neurons induces kisspeptin release in the presence of sex steroids. This has led us to believe that NKB stimulates LH secretion in a kisspeptin-dependent manner despite the latter mechanism having been investigated in the persistent hypogonadal state [e.g. Kiss1r knock out (KO) mice and agonadal monkeys]. Here, we aimed to assess whether in the presence of estradiol, senktide, a NK3R specific agonist, can stimulate LH release in a kisspeptin-independent manner. Adult WT female mice were ovariectomized (OVX) or OVX and estradiol-treated (OVX+E2) and studied in parallel to Kiss1KO or Kiss1KO+E2 littermates (n=10/group). Intracerebroventricular (icv) injections of senktide (600pmol) were performed and blood samples were collected before and 25 min after icv injection. Interestingly, the presence of E2 in Kiss1KO females resulted in LH increase like that observed in WT animals. Subsequently, we aimed to locate the brain area which senktide may be acting to stimulate LH release in Kiss1KO+E2 mice. We mapped the distribution of NK3R in the mouse hypothalamus and moreover investigated the anatomical relationship of NK3R and GnRH expression with dual-label immunohistochemistry as a likely pathway of the kisspeptin-independent stimulation of LH release exerted by NKB. We observed no instances of co-expression between NK3R and GnRH cell bodies (>100 cells analyzed from a total of 16 mice) and only minimal close appositions between the two proteins in the ARC and medial septum. Thus, the kisspeptin-independent action of NKB cannot be attributed to direct stimulation of NK3R located on GnRH neurons. Interestingly, we observed that NK3R immunopositive cells in the posterodorsal medial amygdala (MePD), unlike the ARC, were upregulated in the presence of E2. Next, WT OVX+E2 and Kiss1KO+E2 females were injected with senktide (600pmol; n=5/group) in the ARC, the AVPV/PeN or the MePD; areas found to contain NK3R-expressing neurons, and blood samples were collected before and then every 15 min for 45 min. Senktide significantly stimulated LH secretion in all three areas in WT OVX+E2, but only in the MePD of Kiss1KO+E2 mice. Conversely, in the absence of E2, Kiss1KO females injected with senktide in the MePD did not show any alteration in LH, mimicking the results we obtained after icv administration. These data demonstrate that activation of NK3R in the amygdala stimulates LH release in a kisspeptin independent manner in the presence of estrogen. This novel pathway for LH release, that bypasses the kisspeptin neuron, may play an important role in linking amygdala regulated social cues with gonadotropin release.

Neurokinin-3 receptor activation selectively prolongs atrial refractoriness by inhibition of a background K+ channel

The cardiac autonomic nervous system (ANS) controls normal atrial electrical function. The cardiac ANS produces various neuropeptides, among which the neurokinins, whose actions on atrial electrophysiology are largely unknown. We here demonstrate that the neurokinin substance-P (Sub-P) activates a neurokinin-3 receptor (NK-3R) in rabbit, prolonging action potential (AP) duration through inhibition of a background potassium current. In contrast, ventricular AP duration was unaffected by NK-3R activation. NK-3R stimulation lengthened atrial repolarization in intact rabbit hearts and consequently suppressed arrhythmia duration and occurrence in a rabbit isolated heart model of atrial fibrillation (AF). In human atrial appendages, the phenomenon of NK-3R mediated lengthening of atrial repolarization was also observed. Our findings thus uncover a pathway to selectively modulate atrial AP duration by activation of a hitherto unidentified neurokinin-3 receptor in the membrane of atrial myocytes. NK-3R stimulation may therefore represent an anti-arrhythmic concept to suppress re-entry-based atrial tachyarrhythmias, including AF.

Estradiol alters body temperature regulation in the female mouse

ABSTRACTHot flushes are due to estrogen withdrawal and characterized by the episodic activation of heat dissipation effectors. Recent studies (in humans and rats) have implicated neurokinin 3 (NK3) receptor signaling in the genesis of hot flushes. Although transgenic mice are increasingly used for biomedical research, there is limited information on how 17β-estradiol and NK3 receptor signaling alters thermoregulation in the mouse. In this study, a method was developed to measure tail skin temperature (TSKIN) using a small data-logger attached to the surface of the tail, which, when combined with a telemetry probe for core temperature (TCORE), allowed us to monitor thermoregulation in freely-moving mice over long durations. We report that estradiol treatment of ovariectomized mice reduced TCORE during the light phase (but not the dark phase) while having no effect on TSKIN or activity. Estradiol also lowered TCORE in mice exposed to ambient temperatures ranging from 20 to 36°C. Unlike previous studies in the rat, estradiol treatment of ovariectomized mice did not reduce TSKIN during the dark phase. Subcutaneous injections of an NK3 receptor agonist (senktide) in ovariectomized mice caused an acute increase in TSKIN and a reduction in TCORE, consistent with the activation of heat dissipation effectors. These changes were reduced by estradiol, suggesting that estradiol lowers the sensitivity of central thermoregulatory pathways to NK3 receptor activation. Overall, we show that estradiol treatment of ovariectomized mice decreases TCORE during the light phase, reduces the thermoregulatory effects of senktide and modulates thermoregulation differently than previously described in the rat.

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Objectives Despite intensive research, the pathogenesis of PE remains a mystery. Our study demonstrates a novel interaction between C-reactive protein (CRP) and Neurokinin B (NKB) leading to enhanced activation of the Neurokinin 3 receptor (NK3R). Methods Patient sera and placental tissue were obtained from Memorial Hermann Hospital, Houston, TX. CRP and NKB levels were quantified by ELISA and western blot in term sera and placentas. Coimmunofluorescence was used to colocalize CRP and NKB in placental trophoblast cells. To model PE, pregnant C57BL/6 mice were injected with CRP or with a NK3R antagonist, SB222200, on E13/14. Blood pressure was monitored via tail cuff. Proteinuria was measured via metabolic cage and ELISA on E17. Further verification of the role of the kinin system was undertaken by an in vivo siRNA knockdown of the NK3R and an enzyme involved in phosphocholination of NKB, PCYT1b. Results CRP, an acute phase reactant, was elevated 13-fold in sera and 2-fold in placentas of PE patients. To determine the role of elevated CRP in PE, we injected CRP into pregnant mice. Injection of CRP induced hypertension and proteinuria, indicating its direct role in PE. CRP is known to bind to phosphocholine on damaged cell membranes. NKB, a placentally-enriched peptide and PE-related pathogenic molecule, has been shown to be phosphocholinated, increasing its stability and enhancing NK3R activation. CRP and NKB were found to co-localize to the membrane of human placental trophoblast cells in terminal villi. Finally, in vivo evidence showed nanoparticle-encapsulated siRNA knockdown of NK3R and PCYT1b, and pharmacologic antagonism of NK3R significantly reduced CRP-induced hypertension (159.5 mmHg mean systolic CRP vs. ⩽139 mmHg CRP + treatment groups) and proteinuria in pregnant mice. Conclusions Our findings demonstrate that elevated CRP and NKB/NK3R signaling contribute to disease development. These findings reveal novel therapeutic targets and pathogenic biomarkers for PE. Disclosures N. Parchim: None. W. Wang: None. T. Iriyama: None. O.A. Ashimi: None. C. Liu: None. A.H. Siddiqui: None. S. Blackwell: None. B. Sibai: None. R.E. Kellems: None. Y. Xia: None.

Neurokinin-3 receptor activation in the retrochiasmatic area is essential for the full pre-ovulatory luteinising hormone surge in ewes.

Neurokinin B (NKB) is essential for human reproduction and has been shown to stimulate luteinising hormone (LH) secretion in several species, including sheep. Ewes express the neurokinin-3 receptor (NK3R) in the retrochiasmatic area (RCh) and there is one report that placement of senktide, an NK3R agonist, therein stimulates LH secretion that resembles an LH surge in ewes. In the present study, we first confirmed that local administration of senktide to the RCh produced a surge-like increase in LH secretion, and then tested the effects of this agonist in two other areas implicated in the control of LH secretion and where NK3R is found in high abundance: the preoptic area (POA) and arcuate nucleus (ARC). Bilateral microimplants containing senktide induced a dramatic surge-like increase in LH when given in the POA similar to that seen with RCh treatment. By contrast, senktide treatment in the ARC resulted in a much smaller but significant increase in LH concentrations suggestive of an effect on tonic secretion. The possible role of POA and RCh NK3R activation in the LH surge was next tested by treating ewes with SB222200, an NK3R antagonist, in each area during an oestradiol-induced LH surge. SB222200 in the RCh, but not in the POA, reduced the LH surge amplitude by approximately 40% compared to controls, indicating that NK3R activation in the former region is essential for full expression of the pre-ovulatory LH surge. Based on these data, we propose that the actions of NKB in the RCh are an important component of the pre-ovulatory LH surge in ewes.

Activation of the neurokinin 3 receptor promotes filopodia growth and sprouting in rat embryonic hypothalamic cells.

Members of the tachykinin family have trophic effects on developing neurons. The tachykinin neurokinin 3 receptor (NK3R) appears early in embryonic development; during the peak birthdates of hypothalamic neurons, but its involvement in neural development has not been examined. To address its possible role, immortalized embryonic hypothalamic neurons (CLU209) were treated with CellMask, a plasma membrane stain, or the membranes were imaged in CLU209 cells that were transfected with a pEGFP-NK3R expression vector. Nontransfected cells and transfected cells were then treated with senktide, a NK3R agonist, or Dulbecco's Modified Eagle's Medium (DMEM) and time-lapse confocal images were captured for the following 30 min. Compared to DMEM, senktide treatment led to filopodia initiation from the soma of both nontransfected and transfected CLU209 cells. These filopodia had diameters and lengths of approximately 200 nm and 3 µm, respectively. Pretreatment with an IP3 receptor blocker, 2-aminoethoxydiphenyl borate (2-APB), prevented the senktide-induced growth in filopodia; demonstrating that NK3R-induced outgrowth of filopodia likely involves the release of intracellular calcium. Exposure of transfected CLU209 cells to senktide for 24 h led to further growth of filopodia and processes that extended 10-20 µm. A mathematical model, composed of a linear and population model was developed to account for the dynamics of filopodia growth during a timescale of minutes. The results suggest that the ligand-induced activation of NK3R affects early developmental processes by initiating filopodia formation that are a prerequisite for neuritogenesis.

Activation of Neurokinin 3 Receptors Stimulates GnRH Release in a Location-Dependent but Kisspeptin-Independent Manner in Adult Mice

GnRH neurons form the final common pathway for the central control of reproduction. GnRH release occurs from terminals in the external layer of the median eminence (ME) for neuroendocrine control of the pituitary, and near GnRH-GnRH fiber appositions within the preoptic area (POA). Whether or not control of GnRH secretion by neuromodulators is different in these 2 areas is unknown. Mutations in neurokinin B (NKB) or the neurokinin-3 receptor (NK3R) are linked to hypogonadotropic hypogonadism in humans, suggesting that NKB may regulate GnRH secretion. Using fast scan cyclic voltammetry through carbon-fiber microelectrodes, we examined real-time GnRH release in response to the NK3R agonist senktide in the ME and POA. Coronal brain slices were acutely prepared from adult gonad-intact GnRH-green fluorescent protein male mice, and carbon-fiber microelectrodes were placed either within green fluorescent protein-positive terminal fields of the ME or near GnRH-GnRH fiber appositions in the POA. Senktide induced GnRH release consistently in the ME but not the POA, indicating that GnRH release is differentially regulated by NKB in a location-dependent manner. Senktide also induced GnRH secretion in the ME of kisspeptin-knockout (Kiss1 knockout) mice. Interestingly, release amplitude was lower compared with wild-type mice. These data indicate regulation of GnRH release by NK3R agonists is site specific and suggest that kisspeptin is not a required mediator between NK3R activation and GnRH secretion in the ME. This information will be useful for informing future models of afferent regulation of GnRH release.

Modulation of body temperature and LH secretion by hypothalamic KNDy (kisspeptin, neurokinin B and dynorphin) neurons: A novel hypothesis on the mechanism of hot flushes

Despite affecting millions of individuals, the etiology of hot flushes remains unknown. Here we review the physiology of hot flushes, CNS pathways regulating heat-dissipation effectors, and effects of estrogen on thermoregulation in animal models. Based on the marked changes in hypothalamic kisspeptin, neurokinin B and dynorphin (KNDy) neurons in postmenopausal women, we hypothesize that KNDy neurons play a role in the mechanism of flushes. In the rat, KNDy neurons project to preoptic thermoregulatory areas that express the neurokinin 3 receptor (NK3R), the primary receptor for NKB. Furthermore, activation of NK₃R in the median preoptic nucleus, part of the heat-defense pathway, reduces body temperature. Finally, ablation of KNDy neurons reduces cutaneous vasodilatation and partially blocks the effects of estrogen on thermoregulation. These data suggest that arcuate KNDy neurons relay estrogen signals to preoptic structures regulating heat-dissipation effectors, supporting the hypothesis that KNDy neurons participate in the generation of flushes.

Regulation of Arcuate Neurons Coexpressing Kisspeptin, Neurokinin B, and Dynorphin by Modulators of Neurokinin 3 and κ-Opioid Receptors in Adult Male Mice

Pulsatile GnRH release is essential to fertility and is modulated by gonadal steroids, most likely via steroid-sensitive afferents. Arcuate neurons coexpressing kisspeptin, neurokinin B (NKB), and dynorphin (KNDy neurons) are steroid-sensitive and have been postulated to both generate GnRH pulses and mediate steroid feedback on pulse frequency. KNDy neurons are proposed to interact with one another via NKB and dynorphin to activate and inhibit the KNDy network, respectively, and thus alter kisspeptin output to GnRH neurons. To test the roles of NKB and dynorphin on KNDy neurons and the steroid sensitivity of these actions, targeted extracellular recordings were made of Tac2(NKB)-GFP-identified neurons from castrate and intact male mice. Single-cell PCR confirmed most of these cells had a KNDy phenotype. The neurokinin 3 receptor (NK3R) agonist senktide increased action potential firing activity of KNDy neurons. Dynorphin reduced spontaneous KNDy neuron activity, but antagonism of κ-opioid receptors (KOR) failed to induce firing activity in quiescent KNDy neurons. Senktide-induced activation was greater in KNDy neurons from castrate mice, whereas dynorphin-induced suppression was greater in KNDy neurons from intact mice. Interactions of dynorphin with senktide-induced activity were more complex; dynorphin treatment after senktide had no consistent inhibitory effect, whereas pretreatment with dynorphin decreased senktide-induced activity only in KNDy neurons from intact but not castrate mice. These data suggest dynorphin-mediated inhibition of senktide-induced activity requires gonadal steroid feedback. Together, these observations support the hypotheses that activation of NK3R and KOR, respectively, excites and inhibits KNDy neurons and that gonadal steroids modulate these effects.

Sensitization of hypervigilance effects of cocaine can be induced by NK3 receptor activation in marmoset monkeys

BackgroundCocaine is a widely abused drug which can result in the establishment of addiction. The neurokinin3-receptor (NK3-R) has been linked to cocaine addiction by genetic, epigenetic, and pharmacological studies suggesting that a cocaine-induced increase in NK3-R signaling may contribute to the establishment of cocaine addiction-related behaviors. MethodsHere we measured cocaine-induced sensitization of vigilance- and locomotor behaviors in marmoset monkeys (Callithrix penicillata) in an open field. ResultsWe found a sensitization of vigilance-related, but not locomotor behaviors after repeated cocaine (7mg/kg, i.p.) treatment. There was a cross-sensitization for scan frequency, but not of glance frequency, both vigilance-related behaviors, after repeated treatment with the NK3-R agonist senktide (0.2mg/kg, i.p.) given for 7 days, after a cocaine challenge (5mg/kg, i.p.). ConclusionsThese data suggest that in marmoset monkeys, repeated cocaine treatment leads to a sensitization of vigilance-related behaviors, which have a prominent role in spontaneously expressed activities in this species, but not of locomotor activity. Repeated activation of NK3-Rs can mimic some of the behavioral sensitization effect and may thus contribute to the establishment of cocaine related behaviors.

Activation of tachykinin, neurokinin 3 receptors affects chromatin structure and gene expression by means of histone acetylation

The tachykinin, neurokinin 3 receptor (NK3R) is a g-protein coupled receptor that is broadly distributed in the nervous system and exerts its diverse physiological actions through multiple signaling pathways. Despite the role of the receptor system in a range of biological functions, the effects of NK3R activation on chromatin dynamics and gene expression have received limited attention. The present work determined the effects of senktide, a selective NK3R agonist, on chromatin organization, acetylation, and gene expression, using qRT-PCR, in a hypothalamic cell line (CLU 209) that expresses the NK3R. Senktide (1nM, 10nM) caused a relaxation of chromatin, an increase in global acetylation of histone H3 and H4, and an increase in the expression of a common set of genes involved in cell signaling, cell growth, and synaptic plasticity. Pretreatment with histone acetyltransferase (HAT) inhibitor (garcinol and 2-methylene y-butylactone), that inhibits p300, p300/CREB binding protein (CBP) associated factor (PCAF), and GCN 5, prevented the senktide-induced increase in expression of most, but not all, of the genes upregulated in response to 1nM and 10nM senktide. Treatment with 100nM had the opposite effect: a reduction in chromatin relaxation and decreased acetylation. The expression of four genes was significantly decreased and the HAT inhibitor had a limited effect in blocking the upregulation of genes in response to 100nM senktide. Activation of the NK3R appears to recruit multiple pathways, including acetylation, and possibly histone deactylases, histone methylases, or DNA methylases to affect chromatin structure and gene expression.

NaV‐igating excitement in the enteric nervous system

Na_V‐igating excitement in the enteric nervous system

Tachykinin NK3 receptor contribution to systemic release of vasopressin and oxytocin in response to osmotic and hypotensive challenge

Activation of the neurokinin 3 receptor (NK3R) by a receptor agonist, hypotension, and hyperosmolarity results in the internalization of NK3R expressed by magnocellular neurons and the release of vasopressin (VP) and oxytocin (OT) into the circulation. The contribution of NK3R activation to the release of VP and OT in response to hyperosmolarity and hypotension was evaluated by measuring the release of both hormones following pretreatment with a selective NK3R antagonist, SB-222200. Freely behaving male rats were given an intraventricular injection of either 0.15 M NaCl or 250, 500, or 1,000 pmol SB-222200, and then were administered an intravenous infusion of 2 M NaCl or 0.15 M NaCl (experiment 1), or a bolus intra injection of 0.15 M NaCl or hydralazine (HDZ), a hypotension-inducing drug (experiment 2). Blood samples were taken from indwelling arterial catheters at various time points for 1-2 h, both before and after treatments. Plasma VP and OT levels were determined by ELISA. Blockade of NK3R did not affect the baseline levels of either hormone. In contrast, pretreatment with SB-222200 significantly reduced ( approximately 60%) or abolished the release of VP and OT, respectively, to 2 M NaCl infusion. HDZ-induced VP and OT release was eliminated by pretreatment with 500 pmol SB-222200. Therefore, NK3R activation contributes significantly to the systemic release of both VP and OT in response to osmotic and hypotensive challenges.

NK-3 receptor activation depolarizes and induces an after-depolarization in pyramidal neurons in gerbil cingulate cortex

The involvement of tachykinins in cortical function is poorly understood. To study the actions of neurokinin-3 (NK3) receptor activation in frontal cortex, whole cell patch clamp recordings were performed from pyramidal neurons in slices of cingulate cortex from juvenile gerbils. Senktide (500 nM), a selective NK3 receptor agonist, induced a transient increase in spontaneous EPSPs in layer V pyramidal neurons, accompanied by a small depolarization (∼4 mV). EPSPs during senktide had a larger amplitude and faster 10–90% rise time than during control. Senktide induced a transient depolarization in layer II/III pyramidal neurons, which often reached threshold for spikes. The depolarization (∼6 mV) persisted in TTX, and was accompanied by an increase in input resistance. Senktide also transiently induced a slow after-depolarization, which appeared following a depolarizing pulse. The slow after-depolarization persisted in TTX. These data suggest that activation of NK3 receptors on layer II/III pyramidal neurons induce post-synaptic depolarization and an after-depolarization, which could be mediated by blockade of a leak potassium conductance and a non-selective cation conductance, respectively.

Regional decreases in NK-3, but not NK-1 tachykinin receptor binding in dystonic hamster ( dtsz) brains

Although the pathophysiology of primary dystonias is currently unknown, it is thought to involve changes in the basal ganglia–thalamus–cortex circuit, particularly activity imbalances between direct and indirect striatal pathways. Substance P, a member of the tachykinin family of neuropeptides, is a major component in the direct pathway from striatum to basal ganglia output nuclei. In the present study quantitative autoradiography was used to examine changes in neurokinin-1 (NK-1) and neurokinin-3 (NK-3) receptors in mutant dystonic hamsters ( dt sz), a well characterized model of paroxysmal dystonia. NK-1 receptors were labeled in 10 dystonic brains and 10 age-matched controls with 3 nM [ 3H]-[Sar 9, Met(O 2) 11]-SP. NK-3 binding sites were labeled in adjacent sections with 2.5 nM [ 3H]senktide. NK-1 binding was found to be unaltered in 27 brain areas examined. In contrast, NK-3 binding was significantly reduced in layers 4 and 5 of the prefrontal (−46%), anterior cingulate (−42%) and parietal (−45%) cortices, ventromedial thalamus (−42%) and substantia nigra pars compacta (−36%) in dystonic brains compared to controls. The latter effects may be particularly relevant in view of evidence that activation of NK-3 receptors on dopaminergic neurons in the substantia nigra pars compacta can increase nigrostriatal dopaminergic activity. Since previous studies indicated that a reduced basal ganglia output in mutant hamsters is based on an overactivity of the direct pathway which also innervates substantia nigra pars compacta neurons, the decreased NK-3 binding could be related to a receptor down-regulation. The present finding of decreased NK-3 receptor density in the substantia nigra pars compacta, thalamic and cortical areas substantiates the hypothesis that disturbances of the basal ganglia–thalamus–cortex circuit play a critical role in the pathogenesis of paroxysmal dystonia.

Intraventricular injection of tachykinin NK-sub-3 receptor agonists suppresses the ingestion of NaCl-associated tastes.

The present experiments evaluated whether a salty taste was required for injections of a neurokinin-3 (NK3) receptor agonist (senktide) to suppress intake or whether senktide would reduce the intake of tastes that are predictive of NaCl. During training, different groups of rats were given access to 1% almond + water, 1% almond + 0.3 M NaCl, or 1% almond + 0.1 M sucrose. On the test day, rats were administered intraventricular injections of either saline or 200 ng senktide and then given access to 1% almond + water. Senktide had no effect on the intake of the water-associated or sucrose-associated almond. In contrast, senktide significantly reduced the intake of NaCl-associated almond. Senktide had no effect on almond intake by water-deprived rats. These results show that activation of NK3 receptors reduces the intake of NaCl and of a neutral taste that is predictive of sodium but not of calories.