Phoneutria nigriventer spider venom induces oedema in rat skin by activation of capsaicin sensitive sensory nerves

doi:10.1016/S0014-2999(97)01387-3

European Journal of Pharmacology

Volume 339, Issues 2–3, 27 November 1997, Pages 223-226

https://doi.org/10.1016/S0014-2999(97)01387-3 Get rights and content

Abstract

Phoneeutria nigriventer venom induces oedema formation when injected in the rat dorsal skin and such oedema is, in part, dependent on the stimulation of tachykinin NK₁ receptors. This study investigated whether Phoneutria nigriventer venom acts directly on tachykinin NK₁ receptors, or indirectly to activate sensory neurones which in turn release a tachykinin NK₁ receptor agonist. The plasma extravasation induced by Phoneutria nigriventer venom (1–10 μg/site) in neonatally capsaicin (8-methyl N-vanillyl-6-nonenamide)-pretreated rats was substantially attenuated (P<0.05) but the response to either the tachykinin NK₁ receptor agonist GR73632 ((δAva[l-Pro⁹, N-MeLeu10] substance P-(7–11) 30 pmol/site) or bradykinin (0.3–3 nmol/site) was not affected. These results indicate that Phoneutria nigriventer venom stimulates sensory nerves indirectly. The lack of effect of capsaicin-pretreatment on the GR73632 and bradykinin responses indicated that the tachykinin NK₁ and bradykinin B₂ receptors remained functional. There was no evidence to suggest that Phoneutria nigriventer venom contains a tachykinin NK₁ receptor agonist.

Introduction

It is well established that acute electrical or chemical (e.g. capsaicin, 8-methyl N-vanillyl-6-nonenamide) stimulation of sensory nerve fibres leads to the release of neuropeptides (e.g. tachykinins such as substance P and also calcitonin gene-related peptide) and local inflammatory effects (Lembeck and Holzer, 1979; Escott and Brain, 1993). By comparison, it is well known that the levels of substance P are substantially and irreversibly reduced in primary sensory neurones when rats are treated with capsaicin at a neonatal stage (Gamse et al., 1980).

Venom from the spider Phoneutria nigriventer has a range of effects on both the central and peripheral nervous systems and on peripheral tissues (Entwistle et al., 1982; Diniz et al., 1990; Lopes-Martins et al., 1994; Costa et al., 1996). Indeed, Phoneutria nigriventer venom potently stimulates inflammatory oedema formation in rabbit and rat skin (Antunes et al., 1992, Antunes et al., 1993; Marangoni et al., 1993; Palframan et al., 1996). Previous work has shown that the Phoneutria nigriventer venom-induced oedema in rats is mediated through tachykinin NK₁ receptor since the oedema is inhibited by the selective tachykinin NK₁ receptor antagonist SR140333 ((S)1-{2-[3(3-4-dichlorophenyl)-1-(3-iso-propoxyphenylacetyl)piperidine-3-yl]ethyl}-4-phenyl-1-azoniabicyclo[2.2.2]octone, chloride), see Palframan et al., 1996). The present study was undertaken to determine whether Phoneutria nigriventer venom acts directly on tachykinin NK₁ receptors, or indirectly by activating sensory neurones which release a tachykinin NK₁ receptor agonist.

Section snippets

Animals and capsaicin desensitisation

The experiments were carried out in Wistar rats of both sexes bred in the Departmental Animal House of the Faculty of Medical Science, UNICAMP (São Paulo, Brazil). Rats were pretreated subcutaneously (s.c.) on the second day of life with capsaicin (50 mg/kg) or the corresponding volume (100 μl) of capsaicin-vehicle (10% ethanol and 10% Tween 80, in 0.9% (w/v) NaCl solution) under ether anaesthesia (Jancsó et al., 1977). The rats were used 12–16 weeks after capsaicin pretreatment at which time

Effect of neonatal capsaicin pretreatment on blood pressure and on oedema formation induced by topical capsaicin

The mean arterial blood pressure in conscious, vehicle-pretreated rats (127±3 mm Hg; n=5) was not significantly different from that of conscious, capsaicin-pretreated rats (125±9 mm Hg; n=6).

Topical administration of 5% capsaicin solution onto the dorsal hind paw skin of vehicle-pretreated rats significantly increased (P<0.05) the oedema formation compared to the response obtained following the administration of vehicle to the contralateral paw (10.5±0.9 and 6.2±0.3 μl, respectively; n=4 each).

Discussion

Various symptoms, including pain and cardiovascular disturbances, are observed when animals and humans are bitten by the armed spider Phoneutria nigriventer a species common in Brazil (Lucas, 1988). In addition to polypeptides of different molecular weights, the venom also contains varying amounts of histamine (0.06%) and 5-hydroxytryptamine (5-HT 0.03–0.24%; Schenberg and Pereira-Lima, 1971). We have previously shown that the oedema formation induced by undialysed Phoneutria nigriventer venom

Acknowledgements

S.D.B. and G.d.N. thank the Wellcome Trust for financial support. S.K.P.C. is the recipient of a grant from FAPESP.

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Activation of tissue kallikrein–kininogen–kinin system in rabbit skin by a fraction isolated from Phoneutria nigriventer (armed spider) venom
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Bradykinin activates peripheral capsaicin-sensitive fibres via a second messenger system
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Citation Excerpt :
Tachykinin release is probably secondary to C fibers activation via TRPV1, since PNV does not directly activate NK receptors (Costa et al., 1997). C fibers may store both substance P and glutamate (De Biasi and Rustioni, 1988), and retrograde release of tachykinins contributes to PNV-induced neurogenic inflammation (Costa et al., 1997, 2000). Both tachykinins and glutamate retrograde release are involved in the PNV-induced hyperalgesic state observed in animal models (Zanchet and Cury, 2003).
Phoneutria nigriventer venom (PNV) is a complex mixture of toxins exerting multiple pharmacological effects that ultimately result in severe local pain at the site of the bite. It has been proposed that the PNV-induced pain is mediated by both peripheral and central mechanisms. The nociception triggered by PNV is peripherally mediated by the activation of B₂, 5-HT₄, NMDA, AMPA, NK₁, and NK₂ receptors, as well as TTXS-Na⁺, ASIC, and TRPV1 channels. The activation of tachykinin, glutamate and CGRP receptors along with the production of inflammatory mediators are, at least partially, responsible for the central component of pain. Despite its well established pro-nociceptive properties, PNV contains some toxins with antinociceptive activity, which have been studied in the last few years. The toxins ω-CNTX-Pn4a, ω-CNTX-Pn2a, ω-CNTX-Pn3a, κ-CNTX-Pn1a, U₇-CNTX-Pn1a, δ-CNTX-Pn1a, and Γ-CNTX-Pn1a from PNV, as well as the semi-synthetic peptide PnPP-19 have been tested in different experimental models of pain showing consistent antinociceptive properties. This review aims to discuss the pro- and antinociceptive actions of PNV and its toxins, highlighting possible mechanisms involved in these apparently dualistic properties.
Group III metabotropic glutamate receptors and transient receptor potential vanilloid 1 co-localize and interact on nociceptors
2012, Neuroscience
Several lines of evidence indicate group III metabotropic glutamate receptors (mGluRs) have systemic anti-hyperalgesic effects. We hypothesized this could occur through modulation of transient receptor potential vanilloid 1 (TRPV1) receptors on nociceptors. To address this question we performed anatomical studies to determine if group III mGluRs were expressed on cutaneous axons and if they co-localized with TRPV1. Immunostaining at the electron microscopic level demonstrated that 22% of unmyelinated axons labeled for mGluR8. Immunostaining at the light microscopic level in lumbar dorsal root ganglia (DRG) demonstrated that 80% and 28% of neurons labeled for mGluR8 or TRPV1, respectively. Of those neurons labeled for mGluR8, 25% labeled for TRPV1; of those labeled for TRPV1, 71% labeled for mGluR8. In behavior studies intraplantar injection of the group III mGluR agonist, L-(+)-2-amino-4-phosphonobutyric acid (L-AP-4: 0.1, 1.0, and 10.0 μM) had no effect on paw withdrawal latency (PWL) to heat in naïve rats but administration of 10 μM L-AP-4 prior to 0.05% capsaicin (CAP), significantly attenuated CAP-induced lifting/licking and reduced flinching behavior. The L-AP-4 effect was specific since administration of a group III antagonist α-methyl-3-methyl-4-phosphonophenylglycine (UBP1112) (100 μM) blocked the L-AP-4 effect on CAP, resulting in behaviors similar to CAP alone. Intraplantar injection of UBP1112 alone did not result in nociceptive behaviors, indicating group III mGluRs are not tonically active. Finally, the anti-hyperalgesic effect of group III in this paradigm was local and not systemic since intraplantar administration of L-AP-4 in one hind paw did not attenuate nociceptive behaviors following CAP injection in the contralateral hind paw. Adenyl cyclase/cyclic AMP/PKA may be the second messenger pathway linking these two receptor families because intraplantar injection of forskolin (FSK, 10 μM) reduced PWL to heat and L-AP-4 reversed this FSK effect. Taken together, these results suggest group III mGluRs can negatively modulate TRPV1 through inhibition of adenyl cyclase and downstream intracellular activity, blocking TRPV1-induced activation of nociceptors.
Participation of peripheral tachykinin NK<inf>1</inf> receptors in the carrageenan-induced inflammation of the rat temporomandibular joint
2009, European Journal of Pain
Citation Excerpt :
In fact, the differential roles that substance P can have in inflammatory situations are highly dependent on the inflammatory model or system under study. For example, substance P can directly act as a pro-inflammatory mediator increasing vascular permeability in the skin microcirculation (Costa et al., 1997, 2006b), but in the absence of this neuropeptide, the inflammatory signs observed in the allergic rat airways can be exacerbated (Franco-Penteado et al., 2005, 2006). In response to C. difficile toxin A – induced intestinal inflammation (or other pathological stimuli), substance P released from sensory neurons and/or macrophages can bind to the NK1 receptor in target cells and activate the NF-κB system, thus resulting in increased synthesis of pro-inflammatory mediators, such as interleukin-8.
Temporomandibular disorders represent one of the major challenges in dentistry therapeutics. This study was undertaken to evaluate the time course of carrageenan-induced inflammation in the rat temporomandibular joint (TMJ) and to investigate the role of tachykinin NK₁ receptors. Inflammation was induced by a single intra-articular (i.art.) injection of carrageenan into the left TMJ (control group received sterile saline). Inflammatory parameters such as plasma extravasation, leukocyte influx and mechanical allodynia (measured as the head-withdrawal force threshold) and TNFα and IL-1β concentrations were measured in the TMJ lavages at selected time-points. The carrageenan-induced responses were also evaluated after treatment with the NK₁ receptor antagonist SR140333. The i.art. injection of carrageenan into the TMJ caused a time-dependent plasma extravasation associated with mechanical allodynia, and a marked neutrophil accumulation between 4 and 24 h. Treatment with SR140333 substantially inhibited the increase in plasma extravasation and leukocyte influx at 4 and 24 h, as well as the production of TNFα and IL-1β into the joint cavity, but failed to affect changes in head-withdrawal threshold. The results obtained from the present TMJ-arthritis model provide, for the first time, information regarding the time course of this experimental inflammatory process. In addition, our data show that peripheral NK₁ receptors mediate the production of both TNFα and IL-1β in the TMJ as well as some of the inflammatory signs, such as plasma extravasation and leukocyte influx, but not the nociceptive component.
Characterization of the mechanisms underlying the inflammatory response to Polistes lanio lanio (paper wasp) venom in mouse dorsal skin
2009, Toxicon
Citation Excerpt :
More recent work has shown that the venom of a Brazilian paper wasp, Polybia paulista, stimulates a potent inflammatory response, including leukocyte infiltration and plasma protein extravasation, in the rat peritoneal cavity (de Paula et al., 2006). The general profile of these venoms is similar to the well-characterized inflammatory activity of spider (Costa et al., 1997; Ospedal et al., 2002) and snake (Farsky et al., 1997; Camargo et al., 2005; Fernandes et al., 2006) venoms in the cutaneous microvasculature of rats or mice. The dialysis of P. l. lanio venom partially attenuated the increase in vascular permeability in mouse dorsal skin, suggesting that dialyzable, low molecular mass components (e.g. histamine) or biologically active tachykinin-like peptides contributed to the venom-mediated inflammatory response.
Stings by Polistes wasps can cause life-threatening allergic reactions, pain and inflammation. We examined the changes in microvascular permeability and neutrophil influx caused by the venom of Polistes lanio a paper wasp found in southeastern Brazil. The intradermal injection of wasp venom caused long-lasting paw oedema and dose-dependently increased microvascular permeability in mouse dorsal skin. SR140333, an NK₁ receptor antagonist, markedly inhibited the response, but the NK₂ receptor antagonist SR48968 was ineffective. The oedema was reduced in capsaicin-treated rats, indicating a direct activation of sensory fibres. Dialysis of the venom partially reduced the oedema and the remaining response was further inhibited by SR140333. Mass spectrometric analysis of the venom revealed two peptides (QPPTPPEHRFPGLM and ASEPTALGLPRIFPGLM) with sequence similarities to the C-terminal region of tachykinin-like peptides found in Phoneutria nigriventer spider venom and vertebrates. Wasp venom failed to release histamine from mast cells in vitro and spectrofluorometric assay of the venom revealed a negligible content of histamine in the usual dose of P. l. lanio venom (1 nmol of histamine/7 μg of venom) that was removed by dialysis. The histamine H₁ receptor antagonist pyrilamine, but not bradykinin B₁ or B₂ receptor antagonists, inhibited venom-induced oedema. In conclusion, P. l. lanio venom induces potent oedema and increases vascular permeability in mice, primarily through activation of tachykinin NK₁ receptors by substance P released from sensory C fibres, which in turn releases histamine from dermal mast cells. This is the first description of a neurovascular mechanism for P. l. lanio venom-mediated inflammation. The extent to which the two tachykinin-like peptides identified here contribute to this neurogenic inflammatory response remains to be elucidated.
Unusual profile of leukocyte recruitment in mice induced by a skin secretion of the tree frog Phyllomedusa hypochondrialis
2007, Toxicon
Phyllomedusa hypochondrialis skin secretion can cause both systemic and local effects. In this study, we describe the pattern of local acute inflammatory response after P. hypochondrialis skin secretion injection. The inflammatory reaction in the mice footpad was analysed, including the leukocyte recruitment into local tissue from the peripheral blood, in a mouse model of tissue injury. We also investigated the release of the cytokines IL-1, IL-6 and TNF-α, chemokines KC and MCP-1 and the eicosanoids LTB 4 and PGE₂ in mice. The present findings support the ability of P. hypochondrialis skin secretion to induce local inflammation. In addition, these skin secretion components play a role in the initial rolling and slowing of recruited leukocytes and the transition from rolling to adhesion. Levels of the proinflammatory cytokine IL-6, chemokines KC and MCP-1 as well as the eicosanoid PGE₂ were significantly increased after injection of a skin secretion of P. hypochondrialis (0.6 μg/30 μl intraplantar), whereas no changes in other parameters were observed. Finally, the mechanisms involved in the local inflammatory process induced by P. hypochondrialis skin secretion is one of the questions of relevance related to the complex pathophysiology induced by this particular secretion and other toxins.
How important are NK<inf>1</inf> receptors for influencing microvascular inflammation and itch in the skin? Studies using Phoneutria nigriventer venom
2006, Vascular Pharmacology
Pain and itch sensations are induced by depolarization of C-fibre nerves and possibly other types of fibres. We have evidence from several species, including mice, that skin plasma extravasation induced by the Phoneutria nigriventer spider venom (PNV) is dependent on tachykinin NK₁ receptors. We have now investigated the itching measured as bouts of scratching in response to intradermal (i.d.) PNV in wildtype (NK₁^+/+) and NK₁ receptor knockout (NK₁^−/−) mice. Mice, either NK₁^+/+ or NK₁^−/−, were given a single i.d. injection (0.05 ml) of test agent or vehicle into the shaved dorsal skin, in the intercostal region, in a randomized way. The bouts of scratching were recorded in a blinded manner for 60 min. Oedema formation was concomitantly assessed by the extravascular accumulation of i.v. injected ¹²⁵I-albumin. The i.d. injection of either substance P (at a high dose of 100 nmol/site), or PNV (0.3–10 μg/site) induced oedema formation in NK₁^+/+ but substantially less was observed in NK₁^−/− mice, as previously reported. PNV also induced scratching, but significantly less scratching was observed in NK₁^−/− compared with NK₁^+/+ mice. In contrast, SP did not induce significant scratching at amounts up to 100 nmol in NK₁^+/+ mice. Experiments with an NK₁ receptor antagonist SR140333 (at doses that blocked PNV-induced oedema) revealed that whilst a local co-injection i.d. (1 nmol) in NK₁^+/+ mice had no effect on PNV (3 μg/site)-induced scratching (18.5 ± 3.7 vs. 14.4 ± 3.5 bouts, mean ± S.E.M., n = 5–7), systemic treatment with SR140333 (120 nmol/kg, i.v.) significantly inhibited scratching (14 ± 3.5 vs. 3.1 ± 1.2 bouts, n = 4–6; P < 0.05). These results indicate that NK₁ receptors are involved in mediating PNV-induced scratching and that the location of the receptors is unlikely to be skin. Thus, a distinct separation between endogenous microvascular and PNV nociceptive NK₁-dependent effects is suggested.

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European Journal of Pharmacology

Abstract

Introduction

Section snippets

Animals and capsaicin desensitisation

Effect of neonatal capsaicin pretreatment on blood pressure and on oedema formation induced by topical capsaicin

Discussion

Acknowledgements

References (20)

Phoneutria nigriventer (armed spider) venom induces increased vascular permeability in rat and rabbit skin in vivo

Toxicon

Activation of tissue kallikrein–kininogen–kinin system in rabbit skin by a fraction isolated from Phoneutria nigriventer (armed spider) venom

Toxicon

The effect of Phoneutria nigriventer (armed spider) venom on arterial blood pressure of anaesthetised rats

Eur. J. Pharmacol.

The purification and amino acid sequence of the lethal neurotoxin Tx1 from the venom of the Brazilian `armed' spider Phoneutria nigriventer

FEBS Lett.

Isolation of a pure toxic polypeptide from the venom of the spider Phoneutria nigriventer and its neurophysiological activity on an insect femur preparation

Toxicon

Multiple tachykinin pools in sensory nerve fibers in the rabbit iris

Neuroscience

Spiders in Brazil

Toxicon

Involvement of capsaicin-sensitive neurones in the haemodynamic effects of exogenous vasoactive peptides: Studies in conscious, adult Long Evans rats treated neonatally with capsaicin

Br. J. Pharmacol.

Bradykinin activates peripheral capsaicin-sensitive fibres via a second messenger system

Agents Actions

Effect of a calcitonin gene-related peptide antagonist (CGRP_8–37) on skin vasodilatation and oedema induced by stimulation of the rat saphenous nerve

Br. J. Pharmacol.

Cited by (23)

Pain modulatory properties of Phoneutria nigriventer crude venom and derived peptides: A double-edged sword

Group III metabotropic glutamate receptors and transient receptor potential vanilloid 1 co-localize and interact on nociceptors

Participation of peripheral tachykinin NK<inf>1</inf> receptors in the carrageenan-induced inflammation of the rat temporomandibular joint

Characterization of the mechanisms underlying the inflammatory response to Polistes lanio lanio (paper wasp) venom in mouse dorsal skin

Unusual profile of leukocyte recruitment in mice induced by a skin secretion of the tree frog Phyllomedusa hypochondrialis

How important are NK<inf>1</inf> receptors for influencing microvascular inflammation and itch in the skin? Studies using Phoneutria nigriventer venom

Short communicationPhoneutria nigriventer spider venom induces oedema in rat skin by activation of capsaicin sensitive sensory nerves

Abstract

Introduction

Section snippets

Animals and capsaicin desensitisation

Effect of neonatal capsaicin pretreatment on blood pressure and on oedema formation induced by topical capsaicin

Discussion

Acknowledgements

Toxicon

Toxicon

Eur. J. Pharmacol.

FEBS Lett.

Toxicon

Neuroscience

Toxicon

Involvement of capsaicin-sensitive neurones in the haemodynamic effects of exogenous vasoactive peptides: Studies in conscious, adult Long Evans rats treated neonatally with capsaicin

Br. J. Pharmacol.

Bradykinin activates peripheral capsaicin-sensitive fibres via a second messenger system

Agents Actions

Effect of a calcitonin gene-related peptide antagonist (CGRP8–37) on skin vasodilatation and oedema induced by stimulation of the rat saphenous nerve

Br. J. Pharmacol.

Short communication
Phoneutria nigriventer spider venom induces oedema in rat skin by activation of capsaicin sensitive sensory nerves

Effect of a calcitonin gene-related peptide antagonist (CGRP_8–37) on skin vasodilatation and oedema induced by stimulation of the rat saphenous nerve