19 entries found for : it-63
Goodman RL, He W, Lopez JA, Bedenbaugh MN, McCosh RB, Bowdridge EC, Coolen LM, Lehman MN, Hileman SM (2019) Evidence that the LH surge in ewes involves both neurokinin B–dependent and –independent actions of kisspeptin. Endocrinology 160(12):2990-3000. doi: 10.1210/en.2019-00597Objective: To determine if NKB is involved in the RCh of the ewe in the LH surge.
Summary: NKB signaling in the RCh increases kisspeptin levels critical for the full amplitude of the LH surge in the ewe, but kisspeptin release occurs independently of retrochiamatic area (RCh) input at the onset of the surge to initiate GnRH secretion.
Usage: Bilaterial injections in the RCh of either NK3-SAP or Blank-SAP.
Related Products: NKB-SAP (Cat. #IT-63), Blank-SAP (Cat. #IT-21)
Selective role of neurokinin B in IL-31–induced itch response in mice.
Sakata D, Uruno T, Matsubara K, Andoh T, Yamamura K, Magoshi Y, Kunimura K, Kamikaseda Y, Furue M, Fukui Y (2019) Selective role of neurokinin B in IL-31–induced itch response in mice. J Allergy Clin Immunol 144(4):1130-1133. doi: 10.1016/j.jaci.2019.06.031
Objective: To examine the physiological significance of neurokinin B in IL-31–induced itch sensation.
Summary: IL-31–induced scratching was unaffected by intrathecal injection of Nppb-SAP. In contrast,treatment with Bombesin-SAP reduced IL-31–induced scratching. Neurokinin B acts upstream of GRP to transmit IL-31–induced itch sensation.
Usage: Intrathecal injection
Related Products: Bombesin-SAP (Cat. #IT-40), Nppb-SAP (Cat. #IT-69), Blank-SAP (Cat. #IT-21), NKB-SAP (Cat. #IT-63)
Krajewski-Hall SJ, Miranda Dos Santos F, McMullen NT, Blackmore EM, Rance NE (2019) Glutamatergic neurokinin 3 receptor neurons in the median preoptic nucleus modulate heat-defense pathways in female mice. Endocrinology 160(4):803-816. doi: 10.1210/en.2018-00934Objective: To characterize the thermoregulatory role of MnPO NK3R neurons in female mice.
Summary: Study suggests that KNDy neurons modulate thermosensory pathways for heat defense indirectly via a subpopulation of glutamatergic MnPO neurons that express NK3R.
Usage: Mice were bilaterally injected with 10 ng NK3-SAP in 100 nL PBS (n = 14) or blank-SAP (n = 8) in the preoptic area adjacent to the MnPO.
Related Products: NKB-SAP (Cat. #IT-63), Blank-SAP (Cat. #IT-21)
Goodman RL, Lopez JA, Bedenbaugh MN, Connors JM, Hardy SL< Hileman SM, Coolen LM, Lehman MN (2018) Evidence that the LH surge in ewes involves both neurokinin B-dependent and -independent actions of kisspeptin. Neuroscience 2018 Abstracts 773.20 / YY14. Society for Neuroscience, San Diego, CA.
Summary: It is generally recognized that kisspeptin plays a key role in induction of the LH surge in sheep and we have reported evidence that neurokinin B (NKB) does so as well. Specifically, disrupting NKB signaling in the retrochiasmatic area (RCh) using either an antagonist to its receptor, NK3R, or lesions of NK3R-containing neurons in the RCh with a saporin conjugate (NK3-SAP) reduced the amplitude of the estrogen-induced LH surge by 50%. Because a KISS1R antagonist (p271) also produced a 50% decrease in surge amplitude, we hypothesized that these two systems are organized in series with NKB actions in the RCh stimulating kisspeptin release. If this is the case, then the combination of NK3R lesions and a KISS1R antagonist should produce the same inhibition as either treatment alone. This experiment tested this prediction using a 2 x 2 design. Breeding season ewes were ovariectomized and immediately given an estradiol (E) implant sc and two progesterone implants (CIDRs) intravaginally that produced luteal phase levels of these steroids. Ewes then received bilateral injections of either NK3-SAP (n=6) or Blank-SAP (n=5) into the RCh. Three weeks later, an artificial follicular phase was produced by inserting four 3 cm long E implants 24 hrs after CIDR removal and either saline or p271 was infused into the lateral ventricle for 16-24 hrs after E implantation; LH was monitored every 2-4 hrs for two days. CIDRs were then reinserted and the protocol repeated in a cross-over design so that all ewes received saline and p271 treatment. In Blank-SAP ewes, p271 decreased the peak of the LH surge from 61.2 ± 7.6 to 27.4 ± 4.6 ng/mL and delayed it 8 hrs (from 26.5 ± 0.5 to 34.1 ± 1.2 hrs post E implantation). The NK3-SAP injections alone decreased the peak of the LH surge to 29.7 ± 10.7 ng/mL compared to Blank-SAP, but the peak was not further inhibited by p271 in these NK3-SAP-treated ewes (24.4 ± 1.4 ng/mL). However, p271 delayed the peak of the LH surge (from 28.8 ± 1.2 to 34.8 ± 2.1 hrs post E implantation) in the ewes injected with NK3-SAP. Based on these results, we propose that kisspeptin has two roles in the LH surge in ewes: it initiates the surge independent of NKB signaling in the RCh, and maintains LH secretion during the surge by a NKB-dependent system.
Related Products: NKB-SAP (Cat. #IT-63), Blank-SAP (Cat. #IT-21), Custom Conjugates
Krajewski-Hall SJ, Blackmore EM, McMinn JR, McMullen NT, Rance NE (2017) Increased core temperature following ablation of neurokinin 3 receptor-expressing neurons in the mouse median preoptic nucleus and adjacent preoptic area (MnPO/POA). Neuroscience 2017 Abstracts 414.02 / PP19. Society for Neuroscience, Washington, DC.
Summary: We have previously proposed that KNDy neurons play a role in the generation of hot flushes via neurokinin 3 receptor (NK3R) signaling in the preoptic hypothalamus. This hypothesis is strongly supported by recent clinical studies showing that the number and severity of hot flushes is reduced by treatment with NK3R antagonists. To determine if preoptic NK3R neurons modulate thermoregulation in the mouse, we selectively ablated them using injections of saporin conjugated to a NK3R agonist (NK3-SAP). NK3-SAP was stereotaxically injected into Tacr3-EGFP mice to target the MnPO/POA. Controls received injections of BLANK-SAP. The mice were ovariectomized (OVX) and a telemetry probe was implanted i.p. to measure core temperature (TCORE) and activity. Skin temperature (TSKIN) was monitored using a temperature data-logger attached to the surface of the tail. In experiment 1, circadian temperature rhythms were monitored over a 3 day period in mice housed in their home cages (12 light:12 dark). In experiment 2, mice were exposed to three temperatures, 18, 28 and 35oC, in an environmental chamber. Mice were then implanted s.c. with estradiol (E2) capsules and the experiments repeated. We verified by immunohistochemistry and quantitative microscopy that approximately 80% of the EGFP-NK3R neurons in the MnPO were ablated using NK3-SAP. Ablation of NK3R neurons significantly elevated TCORE during the light phase in both OVX and OVX + E2 mice (OVX: BLANK-SAP, 36.7 + 0.1 vs NK3-SAP 37.4 + 0.1; OVX+E2: Blank-SAP 36.1 + 0.1 vs NK3-SAP 36.8 + 0.1). NK3-SAP injections had no significant effect on TCORE during the dark phase. Ablation of NK3R neurons also increased TCORE during the light phase in mice exposed to 18oC and 28oC. All mice exhibited hyperthermia at 35oC. In contrast, ablation of NK3R neurons in the MnPO/POA had no effect on TSKIN or activity regardless of experimental treatment. These data suggest that NK3R neurons in the MnPO/POA participate in the thermoregulatory axis by promoting heat loss during the day and provide further insight into the CNS thermoregulatory pathways that may be activated during the generation of hot flushes.
Related Products: NKB-SAP (Cat. #IT-63)
Mittelman-Smith M, Krajewski-Hall S, McMullen N, Rance N (2016) Ablation of KNDy neurons results in hypogonadotropic hypogonadism and amplifies the steroid-induced LH surge in female rats. Endocrinology 157:2015-2027. doi: 10.1210/en.2015-1740
Summary: KNDy neurons are a subpopulation of neurons in the infundibular nucleus that coexpress estrogen receptor α, kisspeptin, and neurokinin B (NKB) mRNA. Previous work indicated that altered signaling from KNDy neurons may play a role in the low levels of circulating sex steroids found in hypogonadotropic hypogonadism. Rats received bilateral 10-ng injections of NK3-SAP (Cat. #IT-63) dorsal to the arcuate nucleus. Blank-SAP (Cat. #IT-21) was used as control. In animals with intact ovaries the NK3-SAP lesion resulted in hypogonadotropic hypogonadism. In contrast, the LH surge in lesioned ovariectomized rats was 3-fold higher, demonstrating that KNDy neurons are integral for the control of serum LH levels, estrous cyclicity, and may also have some control over the magnitude of the LH surge.
Related Products: NKB-SAP (Cat. #IT-63), Blank-SAP (Cat. #IT-21)
Helena C, Toporikova N, Kalil B, Stathopoulos A, Pogrebna V, Carolino R, Anselmo-Franci J, Bertram R (2015) KNDy neurons modulate the magnitude of the steroid-induced luteinizing hormone surges in ovariectomized rats. Endocrinology 156:4200-4213. doi: 10.1210/en.2015-1070
Summary: Maturation and reproductive function in mammals is controlled by the kisspeptin neuropeptide. Kisspeptin modulates numerous systems within this framework including the mediation of positive and negative feedback effects of estradiol on luteinizing hormone (LH). In the rat, two kisspeptin neuronal populations exist; one in the anteroventral periventricular nucleus (AVPV), and the KNDy (kisspeptin/ neurokinin B/dynorphin) neurons in the arcuate nucleus. In this work the authors examine the role of KNDy neurons in estradiol positive feedback effects by administering 10-ng bilateral injections of NK3-SAP (Cat. #IT-63) into the arcuate nucleus of rats. The results indicate that KNDy neurons use dynorphin to inhibit AVPV neurons, establishing a regulatory mechanism for the amplitude of steroid-induced LH surges.
Related Products: NKB-SAP (Cat. #IT-63)
Mittelman-Smith M, Krajewski-Hall S, McMullen N, Rance N (2015) Neurokinin 3 receptor-expressing neurons in the median preoptic nucleus modulate heat-dissipation effectors in the female rat. Endocrinology 156:2552-2562. doi: 10.1210/en.2014-1974
Summary: Kisspeptin and Neurokinin B (NKB) expression in the infundibular, or arcuate, nucleus is increased after menopause. Here the authors investigate whether KNDy (kisspeptin, NKB, and dynorphin expressing) neurons are able to influence cutaneous vasodilation through Neurokinin 3 (NK3)-expressing projections from the median preoptic nucleus (MnPO). Rats received two 10-ng injections of NK3-SAP (Cat. #IT-63) into the MnPO. Blank-SAP (Cat. #IT-21) was used as a control. The data indicate that NK3-expressing neurons in the MnPO facilitate vasodilation.
Related Products: NKB-SAP (Cat. #IT-63), Blank-SAP (Cat. #IT-21)
Involvement of kndy neurons in luteinizing hormone surges induced by steroids
Helena CV, Toporikova N, Kalil B, Stathopoulos AM, Anselmo-Franci JA, Bertram R (2014) Involvement of kndy neurons in luteinizing hormone surges induced by steroids. Neuroscience 2014 Abstracts 543.11. Society for Neuroscience, Washington, DC.
Summary: A subset of hypothalamic arcuate neurons that coexpress kisspeptin, neurokinin B and dynorphin (KNDy neurons) has been postulated to be critical for puberty onset and regulation of luteinizing hormone (LH) secretion. A method for targeted ablation of KNDy neurons was recently developed using the molecular neurotoxin saporin conjugated to the selective NK3R agonist [MePhe7]Neurokinin B (Nk3-SAP). Ovariectomized rats were microinjected bilaterally into the arcuate nucleus with Blank-SAP or Nk3-SAP. One set of rats was transcardiacally perfused 1, 2 or 3 weeks after the injections and immunocytochemistry for kisspeptin was performed in the arcuate nucleus region. The number of KNDy neurons was significantly decreased after 1 week of the toxin injection, however maximal fiber ablation was only achieved 3 weeks after the microinjections. Another group of rats was treated with oil (OVO), estradiol (OVE) or estradiol plus progesterone (OVEP). One week later, rats had their jugular vein cannulated and blood samples were taken at 10am and hourly from 3 until 6pm. Selective ablation of KNDy neurons of OVO rats significantly reduced basal LH levels at all time points studied. Basal LH levels in OVE and OVEP animals did not differ between groups, yet KNDy ablation increased peak LH levels in the afternoon of OVE and OVEP rats. A third group of OVE animals was microinjected with norbinaltorphimine (nor-BNI), a kappa opioid receptor antagonist, directly into the anteroventral periventricular nucleus (AVPV) one hour before the expected LH surge. The blockage of dynorphin receptors intra-AVPV significantly increased the LH surge, similar to the effect of KNDy ablation in OVE rats. Our results suggest that KNDy neurons provide inhibition to AVPV kisspeptin neurons through dynorphin and thus regulate the size of the LH surge induced by estradiol or estradiol plus progesterone.
Related Products: NKB-SAP (Cat. #IT-63)
Habenular kisspeptin modulates fear in the zebrafish.
Ogawa S, Nathan FM, Parhar IS (2014) Habenular kisspeptin modulates fear in the zebrafish. Proc Natl Acad Sci U S A 111(10):3841-3846. doi: 10.1073/pnas.1314184111
Summary: The peptide kisspeptin can be found in several areas of the brain, but its role in regions other than the hypothalamus has not been studied. Zebrafish express kiss1 mRNA which is a conserved ortholog of the mammalian KISSI/KissI making zebrafish a viable model for investigating the role of kisspeptin in various brain systems. Animals received 1 μg of the custom conjugate kiss-SAP (see NK3-SAP, Cat. #IT-63) via an intracranial injection. Blank-SAP (Cat. #IT-21) was used as a control. Reducing Kiss1 immunoreactivity in the habenula and the raphe reduced an invoked fear response, indicating a role for kisspeptin in fear inhibition.
Related Products: Blank-SAP (Cat. #IT-21), NKB-SAP (Cat. #IT-63), Custom Conjugates
Ablation of arcuate KNDy neurons amplifies the LH surge in steroid-primed, ovariectomized rats.
Krajewski-Hall SJ, Mittelman-Smith, Mcmullen NT, Rance NE (2013) Ablation of arcuate KNDy neurons amplifies the LH surge in steroid-primed, ovariectomized rats. Neuroscience 2013 Abstracts 274.01. Society for Neuroscience, San Diego, CA.
Summary: KNDy (kisspeptin, neurokinin B and dynorphin) neurons in the arcuate nucleus play an important role in the reproductive axis. We have developed a method to selectively ablate KNDy neurons in the rat using NK3-SAP, a neurokinin 3 receptor agonist conjugated to saporin (Mittelman-Smith, Endocrinology 2012). Ablation of KNDy neurons results in cessation of estrous cycles, ovarian atrophy, a decrease in tonic LH secretion and loss of the rise of serum LH after ovariectomy. Given these profound effects, we tested if we could induce an LH surge in KNDy-ablated rats using a well-established steroid replacement regimen. Rats were maintained on a 14:10 light cycle (lights on at 0500). Using stereotactic surgery, NK3-SAP or Blank-SAP was injected in the arcuate nucleus and rats were allowed to recover for one month before ovariectomy. Seven days after ovariectomy they were implanted with silastic capsules containing 17β-estradiol. Two days later, they were implanted with progesterone capsules (~0830h). Rats were sacrificed in the afternoon at a time previously shown to exhibit peak LH surge levels (~1600h) and the brains were processed for immunohistochemistry. Ablation of KNDy neurons was verified by near complete loss of NKB-immunoreactive neurons in the arcuate nucleus in NK3-SAP rats. At 0830h, tonic levels of serum LH were significantly lower in KNDy ablated rats, consistent with our previous studies. Unexpectedly, the surge in serum LH at 1600h was more than 3-fold higher in NK3-SAP-treated rats compared to Blank-SAP controls (53.5 + 16.5 ng/ml vs 16.5 + 2.1 ng/ml, respectively). To determine if this change was associated with increased activation of GnRH neurons, dual-labeled GnRH-fos immunocytochemistry was performed in rostral hypothalamic sections. There was no significant difference in the total number of GnRH cells counted in 4 matched sections (NK3-SAP, 85.8 + 9.8 vs Blank-SAP, 84.9 + 4.6) or in the percentage of GnRH cells that were activated, as measured by GnRH-fos coexpression (NK3-SAP, 22.4 + 1.8% vs Blank-SAP, 16.6 + 4.9%). In addition, there was no difference between NK3-SAP and Blank-SAP controls in the number of fos-ir cells counted in the AVPV. These data indicate that arcuate KNDy neurons are not required for the induction of an LH surge. The marked increase in the LH surge in KNDy-ablated rats, however, suggests that KNDy neurons are important for regulating the magnitude of the surge.
Related Products: NKB-SAP (Cat. #IT-63)
Helena CV, Kalil B, Anselmo-Franci JA, Bertram R (2013) Time course study of targeted ablation of KNDy neurons, but not tyrosine-hydroxylase neurons, in the rat arcuate nucleus using a neurokinin b-saporin. Endocr Rev 34:OR47-5. 95th Annual Meetin and Expo, San Francisco. doi: 10.1093/edrv/34.supp.1
Summary: Ovariectomized rats were given bilateral injections of NK3-SAP (Cat. #IT-63) into the arcuate nucleus for a time course study of KNDy neuron loss. Blank-SAP (Cat. #IT-21) was used as a control.
Related Products: NKB-SAP (Cat. #IT-63), Blank-SAP (Cat. #IT-21)
Mittelman-Smith MA, Williams H, Krajewski-Hall SJ, McMullen NT, Rance NE (2012) Role for kisspeptin/neurokinin B/dynorphin (KNDy) neurons in cutaneous vasodilatation and the estrogen modulation of body temperature. Proc Natl Acad Sci U S A 109(48):19846-19851. doi: 10.1073/pnas.1211517109
Summary: Menopause is marked by estrogen withdrawal, and also by hot flushes. Given the fact that hypothalamic levels of kisspeptin/neurokinin B/dynorphin (KNDy) neurons are significantly altered in menopause, the authors investigated whether these neurons are involved in the generation of flushes. Rats received bilateral injections of NK3-SAP (Cat. #IT-63) into the arcuate nucleus – a total of 40 ng. Blank-SAP (Cat. #IT-21) was used as control. The data indicate that KNDy neurons promote cutaneous vasodilation, and play a role in 17β-estradiol modulation of body temperature, supporting the hypothesis that these neurons could play a role in the generation of hot flushes.
Related Products: NKB-SAP (Cat. #IT-63), Blank-SAP (Cat. #IT-21)
Krajewski-Hall SJ, Mittelman-Smith MA, Williams H, Lafrance KJ, Mcmullen NT, Rance NE (2012) A role for kisspeptin/neurokinin B/dynorphin (KNDy) neurons in the regulation of estrous cycles and the estrogen modulation of body temperature. Neuroscience 2012 Abstracts 585.02. Society for Neuroscience, New Orleans, LA.
Summary: We have recently described a method to selectively ablate kisspeptin/neurokinin B/dynorphin (KNDy) neurons using stereotaxic injections of NK3-SAP, a neurokinin 3 receptor agonist conjugated to saporin (Mittelman-Smith, Endocrinology, 2012). These studies revealed a critical role for arcuate KNDy neurons in tonic gonadotropin secretion, the rise in serum LH after ovariectomy and estrogen modulation of body weight. Here we determine the effects of KNDy neuron ablation on estrous cycles and the estradiol modulation of body temperature. In the first study, stereotaxic injections of NK3-SAP or Blank-SAP were made in the arcuate nucleus of ovary-intact, adult female rats. Rats with nearly complete KNDy-neuron ablation (verified by NKB immunohistochemistry) exhibited constant diestrus and ovarian atrophy, confirming the importance of these neurons in reproductive regulation. In a second experiment, we evaluated the effects of KNDy neuron ablation on the thermoregulatory axis in rats that were ovariectomized (OVX) and then treated with 17β-estradiol (E2). Tail skin temperatures (TSKIN) and core temperatures (TCORE) were recorded in rats throughout the light/dark cycle and during exposure to different ambient temperatures (TAMBIENT) in an environmental chamber. Notably, the average TSKIN of KNDy-ablated rats was consistently lower than control rats, indicative of lower levels of cutaneous vasodilatation. Moreover, KNDy neuron ablation blocked the reduction of TSKIN by E2 that occurred during the light phase in the environmental chamber, but did not affect the E2 suppression of TSKIN during the dark phase. At a high TAMBIENT of 33 C, the mean TCORE of OVX control rats increased to 39.0 C, and was reduced by E2 replacement. In contrast, at this high TAMBIENT, the average TCORE of OVX, KNDy-ablated rats was lower than OVX control rats, and TCORE was not altered by E2 replacement. Because KNDy neurons exhibit dramatic changes in morphology and gene expression in postmenopausal women, we have hypothesized these neurons contribute to the generation of hot flushes. These studies support this hypothesis by providing the first evidence that KNDy neurons participate in the E2 modulation of body temperature and promote cutaneous vasodilatation, one of the cardinal signs of a hot flush.
Related Products: NKB-SAP (Cat. #IT-63), Blank-SAP (Cat. #IT-21)
Mittelman-Smith MA, Williams H, Krajewski-Hall SJ, Lai J, Ciofi P, McMullen NT, Rance NE ( 2012 ) Arcuate kisspeptin/neurokinin b/dynorphin (KNDy) neurons mediate the estrogen suppression of gonadotropin secretion and body weight. Endocrinology 153(6):2800-2812 . doi: 10.1210/en.2012-1045
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Rance NE, Mittelman-Smith MA, Krajewski-Hall SJ (2012) Featured Article: Use of a novel saporin conjugate (NK3-SAP) to study the function of neurokinin 3 receptor (NK3r)-expressing kisspeptin/neurokinin B/dynorphin (KNDy) neurons in the rat arcuate nucleus. Targeting Trends 13(2)
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Read the featured article in Targeting Trends.
See Also:
Corboz MR, Rivelli MA, Eckel SP (2010) Bronchoconstrictor effect of the tachykinin NK₃-receptor agonists [MePhe⁷]-neurokinin B and senktide in the isolated guinea pig lung. Exp Lung Res 36(9):509-521. doi: 10.3109/01902141003777582
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Selective agonists for substance P and neurokinin receptors
Drapeau G, D'Orléans-Juste P, Dion S, Rhaleb NE, Rouissi NE, Regoli D (1987) Selective agonists for substance P and neurokinin receptors. Neuropeptides 10:43-54. doi: 10.1016/0143-4179(87)90088-6
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Specific agonists for neurokinin B receptors
Drapeau G, d'Orléans-Juste P, Dion S, Rhaleb NE, Regoli D (1987) Specific agonists for neurokinin B receptors. Eur J Pharmacol 136(3):401-403. doi: 10.1016/0014-2999(87)90313-x
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