Regulation of Nociceptive Glutamatergic Signaling by Presynaptic Kv3.4 Channels in the Rat Spinal Dorsal Horn

Presynaptic voltage-gated K+ (Kv) channels in dorsal root ganglion (DRG) neurons are thought to regulate nociceptive synaptic transmission in the spinal dorsal horn. However, the Kv channel subtypes responsible for this critical role have not been identified. The Kv3.4 channel is particularly important because it is robustly expressed in DRG nociceptors, where it regulates action potential (AP) duration. Furthermore, Kv3.4 dysfunction is implicated in the pathophysiology of neuropathic pain in multiple pain models. We hypothesized that, through their ability to modulate AP repolarization, Kv3.4 channels in DRG nociceptors help to regulate nociceptive synaptic transmission. To test this hypothesis, we investigated Kv3.4 immunoreactivity (IR) in the rat cervical superficial dorsal horn (sDH) in both sexes and implemented an intact spinal cord preparation to investigate glutamatergic synaptic currents from second order neurons in the sDH under conditions that selectively inhibit the Kv3.4 current. We found presynaptic Kv3.4 IR in peptidergic and nonpeptidergic nociceptive fibers of the sDH. The Kv3.4 channel is hypersensitive to 4-aminopyridine and tetraethylammonium (TEA). Accordingly, 50 μm 4-aminopyridine and 500 μm TEA significantly prolong the AP, slow the maximum rate of repolarization in small-diameter DRG neurons, and potentiate monosynaptic excitatory postsynaptic currents (EPSCs) in dorsal horn laminae I and II through a presynaptic mechanism. In contrast, highly specific inhibitors of BK, Kv7, and Kv1 channels are less effective modulators of the AP and have little to no effect on EPSCs. The results strongly suggest that presynaptic Kv3.4 channels are major regulators of nociceptive synaptic transmission in the spinal cord.SIGNIFICANCE STATEMENT Intractable neuropathic pain can result from disease or traumatic injury and many studies have been conducted to determine the underlying pathophysiological changes. Voltage-gated ion channels, including the K+ channel Kv3.4, are dysregulated in multiple pain models. Kv3.4 channels are ubiquitously expressed in the dorsal root ganglion (DRG), where they are major regulators of DRG excitability. However, little is known about the ionic mechanisms that regulate nociceptive synaptic transmission at the level of the first synapse in the spinal cord, which is critical to pain transmission in both intact and pathological states. Here, we show that Kv3.4 channels have a significant impact on glutamatergic synaptic transmission in the dorsal horn, further illuminating its potential as a molecular pain therapeutic target.This work was supported by the Vickie and Jack Farber Foundation (M.C.), the Dean's Transformational Science Award (M.C.), the National Institutes of Health (Grant NS079855 to M.C. and Grant NS079702 to A.C.L.), the Dubbs Fellowship Fund (T.M.), Sigma Xi (GIAR Grant G20141015648241 to T.M.), Autifony Therapeutics, Ltd. (M.C.), and the Luso-American Development Foundation (V.P.). We thank Drs. Matthew Dalva, Melanie Elliott, Ethan Goldberg, David Ritter, and Benjamin Zemel, and members of the Covarrubias laboratory for helpful comments and feedback on previous versions of this manuscript; members of the Dalva laboratory for sharing reagents; and Dr. Bruce Bean for providing helpful tips regarding the effects of Kv channel inhibitors on the AP in the DRG.info:eu-repo/semantics/publishedVersio

Potential benefits of the 19-nor-clerodane trans-dehydrocrotonin on the central nervous system

This study examined the effect of trans-dehydrocrotonin (DCTN), a 19-nor-clerodane diterpene isolated from Croton cajucara Benth (Euphorbiaceae), as analgesic and its effect on the central nervous system (CNS) of rodents using different animal models. The DCTN intraperitoneally exhibited mild analgesic activity on hot-plate test, but exhibited strong antinociceptive activity against acetic acid-induced abdominal writhing and the ED50 was calculated to be 44.88 mg/kg. At higher doses (100 mg/kg) it exhibited mild CNS depressant activities in laboratory animals. Moreover, it has negligible antidepressant activity. After taking consideration of the drug interaction, the DCTN can be used as a potent analgesic agent in case of peripheral algesia, without affecting the CNS. Neste estudo avaliou-se o efeito analgésico do diterpeno 19-nor-clerodano trans-desidrocrotonina (DCTN) isolado de Croton cajucara Benth (Euphorbiaceae), bem como seu efeito no sistema nervoso central utilizando-se diferentes tipos de modelos de animais roedores. A administração intraperitoneal deste diterpeno, no teste da placa quente, revelou sua atividade analgésica moderada. No entanto, no teste de contrações abdominais desencadeadas por ácido acético, a DCTN apresentou forte atividade antinociceptiva com DE50 de 44,88 mg/kg. Doses elevadas de DCTN (100 mg/kg) apresentaram moderada atividade depressiva do sistema nervoso central (SNC), não tendo sido evidenciado ação antidepressiva. Após algumas considerações da ação de DCTN em algesia periférica, concluiu-se que esta substância pode ser utilizada como um potente agente analgésico, sem afetar o SNC

Characterization of woody odorant contributors in copaiba oil (Copaifera multijuga Hayne)

Copaifera multijuga Hayne is one of the Copaifera species from which copaiba oil is extracted. Employed in the composition of anti-inflammatory and antiseptic products used in phytotherapy, it is also used by the fragrance industry as a fixative in perfumes, cosmetics and in products such as soaps. To identify the active aroma compounds in C. multijuga oil bouquet, GC-O-MS using AEDA (Aroma Extract Dilution Analysis) was used after the quantification of the components by GC-FID. The results obtained pointed to minor compounds such as delta-cadinene (1.9%, FD 64), delta-cadinol (0.9%, FD 128), (Z)-alpha-santalol (0.2%, FD 128) caryophyllene oxide (0.2%, FD 64), alpha-cadinol (0.1%, FD 128) and tau-muurolol (0.1%, FD 128) as the most intense compounds in the odor of the copaiba oil studied. Chiral GC-O-MS showed (+)-delta-cadinene as the only enantiomer present in the oil, with a sweet, green and refreshing aroma

Chemical composition and release in situ due to injury of the invasive coral tubastraea (Cnidaria, Scleractinia)

Substâncias químicas de defesa contra consumidores e competidores podem ser usadas por espécies invasoras marinhas como estratégia de colonização e perpetuação em novo ambiente. Entretanto, há poucos estudos experimentais que demonstrem as possíveis interações negativas entre corais escleractínios. Este trabalho tem como objetivo caracterizar os metabólitos secundários dos corais invasores Tubastraea tagusensis e T. coccinea; avaliar através da técnica de amostragem in situ quais são as substâncias de T. tagusensis liberadas na água do mar, com o auxílio de aparelho subaquático com colunas Sep-Paks®. Colônias dos corais invasores Tubastraea spp foram coletadas na Baía de Ilha Grande, RJ, e extraídas com MeOH. Os extratos foram submetidos à eluições com hexano, DCM e MeOH, e analisados por CG-EM. Estearato de metila e palmitato de metila foram as substâncias majoritárias das frações hexânicas e hexano: DCM, enquanto o colesterol foi a substância mais abundante das frações DCM e DCM:MeOH de Tubastraea spp. O material orgânico retido nas colunas Sep-Paks® foi identificado como hidrocarbonetos. Diferenças significativas entre controle e tratamento foram relacionadas a diferentes quantidades de 1-hexadeceno, n-hexadecano e n-eicosano. A produção de substâncias de defesas em Tubastraea spp permite especular sobre a ameaça que estes corais invasores representam para as comunidades bentônicas da Ilha Grande.Defensive chemistry may be used against consumers and competitors by invasive species as a strategy for colonization and perpetuation in a new area. There are relatively few studies of negative chemical interactions between scleratinian corals. This study characterizes the secondary metabolites in the invasive corals Tubastraea tagusensis and T. coccinea and relates these to an in situ experiment using a submersible apparatus with Sep-Paks® cartridges to trap substances released by T. tagusensis directly from the sea-water. Colonies of Tubastraea spp were collected in Ilha Grande Bay, RJ, extracted with methanol (MeOH), and the extracts washed with hexane, dichloromethane (DCM) and methanol, and analyzed by GC/MS. Methyl stearate and methyl palmitate were the major components of the hexane and hexane:MeOH fractions, while cholesterol was the most abundant in the DCM and DCM:MeOH fractions from Tubastraea spp. The organic material retained in Sep-Paks® cartridges was tentatively identified as hydrocarbons. There was a significant difference between treatments and controls for 1-hexadecene, n-hexadecane and n-eicosane contents. The production of defensive substances by the invasive corals may be a threat to the benthic communities of the region, which include endemic species

Cytotoxicity and antibacterial studies of iridoids and phenolic compounds isolated from the latex of Himatanthus sucuuba

The latex of Himatanthus sucuuba (Spruce) Woodson, used popularly in the Amazon for the treatment of tumors, gastritis, inflammations and infections, was evaluated for cytotoxicity and antibacterial activities. The iridoid lactones, plumericin and isoplumericin were isolated from latex by bioassay fractionation and were found to be associated with DNA damage. Gallic acid exhibited the highest antimicrobial activity among the phenolic compounds isolated from the aqueous fraction. The compounds associated to cytotoxicity and antimicrobial activities could be responsible to the effects of this species used in traditional medicine.Key words: Himatanthus sucuuba, iridoids, phenolics, cytotoxicity, antibacterial