Botulinum Toxin and Pain.

This chapter is focused on analgesic mechanism of action of botulinum toxin type A (BoNT-A) including the action beyond peripheral nerve endings. With the exception of the meninges and possibly urinary bladder, the presence of BoNT-A activity in the periphery, cleaving SNAP25 as a target molecule, up to now was not convincingly shown. In contrast many reports demonstrated BoNT-A activity and the presence of cleaved SNAP25 in the brain and spinal cord. In a model of mirror pain BoNT-A analgesic effect can be achieved even without participation of peripheral nerve ending. Thus generalized hypothesis central or peripheral mechanism of action belongs to history, and there is a need to confirm or dispute the results with meninges, urinary bladder, and possibly with other, especially visceral organs.There are two general options for the central actions of BoNT-A: 1. The activity ends by silencing primary sensory neuron thereby stopping the pain information further in the CNS. 2. Or thereafter, indirectly or transsynaptically, BoNT-A triggers smaller or larger neural loops, forming memory of pain in the CNS that could explain the bilateral effects after unilateral peripheral administration, similar effect in mirror image allodynia and the like Intensive research has shown that peripherally administered BoNT-A reaches the CNS by axonal transport. There is increasing evidence that BoNT-A is preventing pain in a growing range of disorders. In the absence of unexpected findings, or an increase in the uncontrolled use of illicit preparations by uneducated persons, BoNT-A is emerging as a new long-lasting and relatively safe analgesic.

Vladimir Sertic: forgotten pioneer of virology and bacteriophage therapy.

Vladimir Sertic was a pioneer of bacteriophage research in the period between the two world wars. He was born and educated in Croatia, where he made his initial discoveries, and joined Félix d'Herelle's Laboratoire du Bactériophage in Paris in 1928. Original documents and a box with hundreds of sealed bacteriophages samples were kept in Sertic's Zagreb home for decades. Following Vladimir's death, his sister passed this archival material to Professor Zdravko Lackovic in 1989. Some years later, these artefacts were opened and studied. Additionally, we conducted a literature search using the term 'Vladimir Sertic' in the databases PubMed and Google Scholar. After a detailed examination of these data, we established a chronology of his work and compiled a list of his scientific publications. A complete bibliography, with the exception of those publications already cited here, is provided as an appendix. Sertic's key contributions included the exploration of the properties of phage lysins, the devising of a uniform bacteriophage classification system and, in collaboration with his protégé, Nikolai Boulgakov, the isolation of numerous bacteriophage strains, including the famous φX174. Finally it was Sertic's pioneering work in Zagreb that offered confirmation that phages are live agents.

Transynaptic Action of Botulinum Neurotoxin Type A at Central Cholinergic Boutons.

Botulinum neurotoxin Type A (BoNT/A) is an effective treatment for several movement disorders, including spasticity and dystonia. BoNT/A acts by cleaving synaptosomal-associated protein of 25 kDa (SNAP-25) at the neuromuscular junction, thus blocking synaptic transmission and weakening overactive muscles. However, not all the therapeutic benefits of the neurotoxin are explained by peripheral neuroparalysis, suggesting an action of BoNT/A on central circuits. Currently, the specific targets of BoNT/A central activity remain unclear. Here, we show that catalytically active BoNT/A is transported to the facial nucleus (FN) after injection into the nasolabial musculature of rats and mice. BoNT/A-mediated cleavage of SNAP-25 in the FN is prevented by intracerebroventricular delivery of antitoxin antibodies, demonstrating that BoNT/A physically leaves the motoneurons to enter second-order neurons. Analysis of intoxicated terminals within the FN shows that BoNT/A is transcytosed preferentially into cholinergic synapses. The cholinergic boutons containing cleaved SNAP-25 are associated with a larger size, suggesting impaired neuroexocytosis. Together, the present findings indicate a previously unrecognized source of reduced motoneuron drive after BoNT/A via blockade of central, excitatory cholinergic inputs. These data highlight the ability of BoNT/A to selectively target and modulate specific central circuits, with consequent impact on its therapeutic effectiveness in movement disorders.SIGNIFICANCE STATEMENT Botulinum neurotoxins are among the most potent toxins known. Despite this, their specific and reversible action prompted their use in clinical practice to treat several neuromuscular pathologies (dystonia, spasticity, muscle spasms) characterized by hyperexcitability of peripheral nerve terminals or even in nonpathological applications (i.e., cosmetic use). Substantial experimental and clinical evidence indicates that not all botulinum neurotoxin Type A (BoNT/A) effects can be explained solely by the local action (i.e., silencing of the neuromuscular junction). In particular, there are cases in which the clinical benefit exceeds the duration of peripheral neurotransmission blockade. In this study, we demonstrate that BoNT/A is transported to facial motoneurons, released, and internalized preferentially into cholinergic terminals impinging onto the motoneurons. Our data demonstrate a direct central action of BoNT/A.

Decompression endoscopic surgery for frontal secondary headache attributed to supraorbital and supratrochlear nerve entrapment: a comprehensive review.

In the last decade, a new surgical treatment modality was developed for frontal secondary headache, based on the assumption that the trigger of this pain entity is the entrapment of peripheral sensory nerves. The surgery entails a procedure, where an endoscopic approach is used to decompress the supraorbital and supratrochlear nerve branches, which are entrapped by the periosteum in the region of the corrugator supercilii muscle. Candidates for the surgery define their headache as moderate to severe persistent daily pressure or tension, localized in the frontal area, sometimes accompanied by symptoms of nausea and photophobia mimicking a primary headache-migraine. We created a step-by-step screening algorithm which is used to differentiate patients that have the highest chance for a successful surgical decompression. Up to now, published data regarding this type of surgery demonstrate long-lasting successful outcomes while adverse effects are minor. This article reviews and discusses from a surgeon's perspective decompression surgery for secondary headache attributed to supraorbital and supratrochlear nerve entrapment.

"Bunanje": XX Century Abuse of Atropa Belladonna Halucinogenic Berries in Continental Croatia.

BACKGROUND: Atropa belladonna (Engl. deadly nightshade, Cro. velebilje, bunika) is a plant containing pharmacologically active, potentially toxic alkaloids: atropine, hyocyamine and scopolamine. The risk of poisoning in children is important because of possible confusion of black/dark blue belladonna fruit berries with other edible berries. There are many reports in literature of accidental intoxication but no report on traditional intentional usage to achieve hallucinogenic effects. SUBJECTS AND METHODS: Here we report purposeful ingestion of Atropa belladonna berries for hallucinatory effects among adolescents in Bjelovar region in north part of Croatia. This has been happening among children/adolescents while they were grazing animals. We visited a dozen villages in the region and spoke to the oldest mostly to the elderly residents. RESULTS: The existence of such abuse of Atropa belladonna berries in the first part of XX century was confirmed by eight narrators from five distinct places in the region. Interestingly this type of behavior had a specific name "bunanje", unknown in Croatian language, but clearly associated with local plant name bun or bunika. According to informants consumers of berries would develop delirium or hallucinations associated behavior, incoherent and meaningless speech. However nobody remembers any severe case of poisoning. At the regional hospital in Bjelovar in the Pediatric department, there is no record of poisoning with Atropa belladonna. To our knowledge this is the first report of intentional consumption of belladonna berries to achieve the hallucinogenic effect. CONCLUSIONS: The fact that the custom was observed in five distinct spots and it had its specific name "bunanje" suggest that those are not isolated random events but the type of practices; seasonal abuse of hallucinogenic berries of Atropa belladonna, among rural adolescents in the first part of XX century.

Comparison of analgesic effects of single versus repeated injection of botulinum toxin in orofacial formalin test in rats.

Long-term effectiveness and repeated administration of botulinum toxin A are the basis for its use in both neuromuscular disorders and certain painful conditions. Botulinum toxin A has been recently approved for migraine treatment, and its off-label use extends to other craniofacial pain disorders. However, recently it was reported that, after repeated injection, botulinum toxin loses its antinociceptive efficacy in rats. In present study with a similar design, we compared the effects of single and repeated injections of botulinum toxin in formalin-induced orofacial pain. No statistically significant differences were found between single or repeatedly treated animal groups. Our results are in line with the clinical experience and suggest that botulinum toxin can be re-administered in orofacial pain treatment.

The safety of botulinum neurotoxin type A's intraarticular application in experimental animals.

In vivo studies of botulinum neurotoxin type A (BoNT-A) enabled characterization of its activity in the nociceptive sensory system separate from its preferred action in motor and autonomic nerve terminals. However, in the recent rodent studies of arthritic pain which employed high intra-articular (i.a.) doses (expressed as a total number of units (U) per animal or U/kg), possible systemic effects have not been conclusively excluded. Herein we assessed the effect of two pharmaceutical preparations, abobotulinumtoxinA (aboBoNT-A, 10, 20, and 40 U/kg corresponding to 0.05, 0.11, and 0.22 ng/kg neurotoxin) and onabotulinumtoxinA (onaBoNT-A, 10 and 20 U/kg corresponding to 0.09 and 0.18 ng/kg, respectively) injected into the rat knee, on safety-relevant readouts: digit abduction, motor performance and weight gain during 14 days post-treatment. The i. a. toxin produced dose-dependent impairment of the toe spreading reflex and rotarod performance, which was moderate and transient after 10 U/kg onaBoNT-A and ≤20 U/kg aboBoNT-A doses, and severe and long-lasting (examined up to 14 days) after ≥20 U/kg of onaBoNT-A and 40 U/kg aboBoNT-A. In addition, lower toxin doses prevented the normal weight gain compared to controls, while higher doses induced marked weight loss (≥20 U/kg of onaBoNT-A and 40 U/kg aboBoNT-A). Commonly employed BoNT-A formulations, depending on the doses, cause local relaxation of the surrounding muscles and systemic adverse effects in rats. Thus, to evade possible toxin unwanted local or systemic spread, careful dosing and motor testing should be mandatory in preclinical behavioral studies, irrespective of the sites and doses of toxin application.

Antinociceptive Actions of Botulinum Toxin A1 on Immunogenic Hypersensitivity in Temporomandibular Joint of Rats.

Botulinum neurotoxin type A1 (BoNT-A) reduces the peripheral peptide and cytokine upregulation in rats with antigen-evoked persistent immunogenic hypersensitivity (PIH) of the temporomandibular joint (TMJ). Herein, we examined the effects of two preparations of BoNT-A, abobotulinumtoxinA (aboBoNT-A; Dysport) and onabotulinumtoxinA (onaBoNT-A; Botox), on spontaneous and evoked nociceptive behaviors, as well as on central neuronal and astroglial activation. The antigen-evoked PIH was induced in rats via repeated systemic and unilateral intra-articular (i.a.) injections of methylated bovine serum albumin (mBSA). Rats were subsequently injected with unilateral i.a. aboBoNT-A (14 U/kg), onaBoNT-A (7 U/kg), or the vehicle (saline). After i.a. treatments, spontaneous and mechanically evoked nocifensive behaviors were assessed before and after the low-dose i.a. formalin (0.5%) challenge. The central effects of BoNT-A were assessed by an immunohistochemical analysis of cleaved synaptosomal-associated protein 25 (cSNAP-25) presence, c-Fos, GFAP, and CGRP expression in the trigeminal nucleus caudalis (TNC). Both BoNT-A preparations similarly reduced the formalin-induced spontaneous pain-related behaviors and mechanical allodynia of the hypernociceptive rats. Likewise, their effects were associated with the central occurrence of cSNAP-25 and reduction of c-Fos and GFAP upregulation in the TNC. BoNT-A antinociceptive activity on the PIH is associated with the toxin axonal transport to trigeminal sensory areas and reduction of neuronal and glial activation in central nociceptive regions.

New analgesic: Focus on botulinum toxin.

In 2010, Kissin concluded pessimistically that of the 59 new drugs introduced in the fifty-year period between 1960 and 2009 and still in use, only seven had new molecular targets. Of these, only one, sumatriptan, was effective enough to lead to the introduction of multiple drugs targeting the same target molecules (triptans) (Kissin, 2010). Morphine and acetylsalicylic acid (aspirin), introduced for the treatment of pain more than a century ago, continue to dominate biomedical publications despite their limited effectiveness in many areas (e.g., neuropathic pain) and serious adverse effects. Today, are we really closer to ideal analgesics that would work hard enough, long enough, and did not have unwanted side effects? The purpose of the present article is to analyze where we are now. Several drugs, like long-acting opioids or botulinum toxins open some hope. Advantage of botulinum toxin A is unique duration of action (months). New discoveries showed that after peripheral application botulinum toxin by axonal transport reaches the CNS. Major analgesic mechanism of action seems to be of central origin. Will botulinum toxin in the CNS bring new indications and or/adverse effects? Much more basic and clinical research should be in front of us. Although relatively safe as a drug, botulinum toxin is not without adverse effect. Policy makers, clinicians and all those applying botulinum toxin should be aware of that. Unfortunately the life without the pain is still not possible.

Association of Intranasal and Neurogenic Dural Inflammation in Experimental Acute Rhinosinusitis.

BACKGROUND: Nasal cavity and sinus disorders, such as allergic rhinitis, rhinosinusitis, or certain anatomical defects, are often associated with transient or ongoing headaches. On the other hand, migraine headache patients often exhibit pain referral over the area of nasal sinuses and typical nasal autonomic symptoms involving congestion and rhinorrhea. Mechanism for convergence of nasal or sinus disorders and headaches is unknown. Herein, we examined the association of sino-nasal inflammatory pain with common preclinical indicators of trigeminovascular system activation such as dural neurogenic inflammation (DNI) and neuronal activation in brainstem nociceptive nuclei. METHODS: Nasal and paranasal cavity inflammation and pain was induced by formalin (2.5%/10 µl) or capsaicin (0.1%/10 µl) instillation at the border of maxillary sinus and nasal cavity in rats. Quantification of inflammation of nasal mucosa and DNI was performed by spectrophotometric measurement of Evans blue - plasma protein complex extravasation. Pain behavior was quantified by rat grimace scale (RGS). Nociceptive neuronal activation in caudal part of spinal trigeminal nucleus (TNC) was assessed by c-Fos protein immunohistochemistry. RESULTS: Capsaicin and formalin administered into rat nasal cavity increased plasma protein extravasation in the nasal mucosa and dura mater. Intensity of plasma protein extravasation in nasal mucosa correlated with extravasation in dura. Similarly, facial pain intensity correlated with nociceptive neuronal c-Fos activation in the TNC. CONCLUSION: Present data show that inflammatory stimuli in deep nasal and paranasal structures provoke distant intracranial changes related to trigeminovascular system activation. We hypothesize that this phenomenon could explain overlapping symptoms and comorbidity of nasal/paranasal inflammatory disorders with migraine.

Single intracerebroventricular injection of botulinum toxin type A produces slow onset and long-term memory impairment in rats.

It is generally believed that the cholinergic system plays an important role in normal cognitive functioning. Botulinum toxin is the most potent toxin of the peripheral cholinergic system and today it is used in the treatment of a variety of neurological disorders. However, it is surprising that its effect on cognitive processes has been investigated in only two publications. Short-term effects of the central application of botulinum toxin (BTX) type B have been associated with cognitive impairment in animals, while results with type A are ambiguous. In the present study, we have investigated the duration of memory impairment after an intracerebroventricular administration of BTX-A in rats. Two experiments were performed, lasting 12 and 5 months, respectively. In both experiments, the same dose of BTX-A was applied (2 U/kg) and the Morris water maze test was used in the assessment of memory performance. Results show that a single icv injection of a small dose of BTX-A significantly impairs the water maze performance. In both experiments, impairment was apparently of a slow onset and long lasting (up to 12 months). The length and pattern of attenuation suggest development of dementia-like deficits. In addition to providing a potentially new experimental model of memory impairment, these results question the idea of an intracranial application of BTX in the treatment of CNS disorders.

Psychoactive drugs and neuroplasticity.

The influence of psychoactive drugs on neuroplasticity, especially on neurogenesis is reviewed. From psychopharmacological point of view most interesting results are those showing neurogenesis that neurogenesis is increased by SSRI. However, the role of serotonin system in neurogenesis as well as significance of neurogenesis in the beneficial effect of psychotropic drugs requires a lot of additional and new inventive research.

[Botulinum toxin type A and cholinergic system]. / Botulinum toksin tipa A i kolinergicni sustav.

Anaerobic bacterium Clostridium botulinum produces seven different serotypes of botulinum neurotoxins (A-G), which specifically act at the peripheral cholinergic nerve terminals blocking the release of acethylcholine. Primary site of action of botulinum toxin type A (BT-A) is neuromuscular end plate where it specifically cleaves SNAP-25, one of the proteins necessary for neuroegzocytosis. The consequence is long-lasting muscle paralysis. Although BT-A is one of the most potent toxins in nature, over the last 20 years, intramuscular injections of nanogram quantities of BT-A have been used to treat various conditions characterized by increased muscle contraction, like dystonias, spasticity related to cerebral palsy etc but also for autonomic nervous system disorders, like hyperhydrosis. Long duration of action (several months) after peripheral application is the most prominent feature of the toxin's action. Although the acute mechanism of action on neuromuscular junction is largely investigated, there are still some unknowns related to: the passage of BT-A through epithelial barriers, specific recognition of peripheral cholinergic neurons. The mechanism of long duration of action, which is the base of therapeutic use of BT-A, is poorly understood.

Involvement of substance P in the antinociceptive effect of botulinum toxin type A: Evidence from knockout mice.

The antinociceptive action of botulinum toxin type A (BoNT/A) has been demonstrated in behavioral animal studies and clinical settings. It was shown that this effect is associated with toxin activity in CNS, however, the mechanism is not fully understood. Substance P (SP) is one of the dominant neurotransmitters in primary afferent neurons transmitting pain and itch. Thus, here we examined association of SP-mediated transmission and BoNT/A antinociceptive action by employing gene knockouts. Antinociceptive activity of intraplantarly (i.pl.) injected BoNT/A was examined in mice lacking the gene encoding for SP/neurokinin A (tac1-/-) or SP-preferred receptor neurokinin 1 (tac1r-/-), compared to control C57Bl/6J wild type animals. BoNT/A action was assessed in inflammatory pain induced by formalin and CFA, and neuropathic pain induced by partial sciatic nerve ligation. BoNT/A activity in CNS was examined by c-Fos and BoNT/A-cleaved SNAP-25 immunohistochemistry. In wild type mice, acute (formalin-evoked) and chronic pain (neuropathic and inflammatory) was reduced by peripherally injected BoNT/A. In tac1-/- and tac1r-/- knockout mice, BoNT/A exerted no analgesic effect. In control animals BoNT/A reduced the formalin-evoked c-Fos expression in lumbar dorsal horn, while in knockout mice the c-Fos expression was not reduced. After peripheral toxin injection, cleaved SNAP-25 occurred in lumbar dorsal horn in all animal genotypes. BoNT/A antinociceptive activity is absent in animals lacking the SP and neurokinin 1 receptor encoding genes, in spite of presence of toxin's enzymatic activity in central sensory regions. Thus, we conclude that the integrity of SP-ergic system is necessary for the antinociceptive activity of BoNT/A.

Activity of botulinum toxin type A in cranial dura: implications for treatment of migraine and other headaches.

BACKGROUND AND PURPOSE: Although botulinum toxin type A (BoNT/A) is approved for chronic migraine treatment, its mechanism of action is still unknown. Dural neurogenic inflammation (DNI) commonly used to investigate migraine pathophysiology can be evoked by trigeminal pain. Here, we investigated the reactivity of cranial dura to trigeminal pain and the mechanism of BoNT/A action on DNI. EXPERIMENTAL APPROACH: Because temporomandibular disorders are highly comorbid with migraine, we employed a rat model of inflammation induced by complete Freund's adjuvant, followed by treatment with BoNT/A injections or sumatriptan p.o. DNI was assessed by Evans blue-plasma protein extravasation, cell histology and RIA for CGRP. BoNT/A enzymatic activity in dura was assessed by immunohistochemistry for cleaved synaptosomal-associated protein 25 (SNAP-25). KEY RESULTS: BoNT/A and sumatriptan reduced the mechanical allodynia and DNI, evoked by complete Freund's adjuvant. BoNT/A prevented inflammatory cell infiltration and inhibited the increase of CGRP levels in dura. After peripheral application, BoNT/A-cleaved SNAP-25 colocalized with CGRP in intracranial dural nerve endings. Injection of the axonal transport blocker colchicine into the trigeminal ganglion prevented the formation of cleaved SNAP-25 in dura. CONCLUSIONS AND IMPLICATIONS: Pericranially injected BoNT/A was taken up by local sensory nerve endings, axonally transported to the trigeminal ganglion and transcytosed to dural afferents. Colocalization of cleaved SNAP-25 and the migraine mediator CGRP in dura suggests that BoNT/A may prevent DNI by suppressing transmission by CGRP. This might explain the effects of BoNT/A in temporomandibular joint inflammation and in migraine and some other headaches.

Botulinum neurotoxin type A: Actions beyond SNAP-25?

Botulinum neurotoxin type A (BoNT/A), the most potent toxin known in nature which causes botulism, is a commonly used therapeutic protein. It prevents synaptic vesicle neuroexocytosis by proteolytic cleavage of synaptosomal-associated protein of 25 kDa (SNAP-25). It is widely believed that BoNT/A therapeutic or toxic actions are exclusively mediated by SNAP-25 cleavage. On the other hand, in vitro and in vivo findings suggest that several BoNT/A actions related to neuroexocytosis, cell cycle and apoptosis, neuritogenesis and gene expression are not necessarily mediated by this widely accepted mechanism of action. In present review we summarize the literature evidence which point to the existence of unknown BoNT/A molecular target(s) and modulation of unknown signaling pathways. The effects of BoNT/A apparently independent of SNAP-25 occur at similar doses/concentrations known to induce SNAP-25 cleavage and prevention of neurotransmitter release. Accordingly, these effects might be pharmacologically significant. Potentially the most interesting are observations of antimitotic and antitumor activity of BoNT/A. However, the exact mechanisms require further studies.

Involvement of the peripheral cholinergic muscarinic system in the compensatory ovarian hypertrophy in the rat.

In the present experiments, unilateral ovariectomy (ULO) induced compensatory hypertrophy (COH) of the remaining rat ovary (60%-85% increase in ovarian weight, total proteins, and total RNA and DNA). An increased thymidine uptake preceded the organ enlargement. COH was inhibited by i.p.-administered muscarinic antagonist propantheline (dose-dependently) or botulinum toxin delivered locally to the ovary. The effects were reversed by bethanecol i.p. (a muscarinic agonist). In sham ULO animals, [3H]-scopolamine binding to ovarian membranes indicated the existence of muscarinic receptors (Kd 2.5 nM, Bmax 12 fmol/mg proteins, Hill 1.0). The ovarian 1,2-diacylglycerol (DAG) was 120-150 pmol/mg tissue and did not react to carbachol in vitro (50 microM). At 15 minutes after ULO, the [3H]-scopolamine binding was unchanged (Kd 2.6 nM, Bmax 12.6 fmol/mg tissue, Hill 1.0), but the ovarian DAG was increased (280-350 pmol/mg tissue) and increased further in response to carbachol (460-550 pmol/mg tissue). After ULO, ovarian DAG remained continuously responsive to carbachol. The ULO-induced DAG increase and enhanced susceptibility to carbachol were inhibited by the botulinum toxin or atropine pretreatments. Abdominal vagotomy done immediately before ULO also inhibited the ULO-induced DAG increase and DAG responsiveness to carbachol. However, when the vagotomy was performed 10 mins after ULO, the ovarian DAG remained responsive to carbachol in vitro. The data suggest that the peripheral cholinergic system, including the ovarian muscarinic receptors, stimulates COH. This is apparently associated with the ULO-induced coupling of the ovarian muscarinic receptors to phosphoinositide (PI) breakdown. Vagus plays a role in the occurrence of the changed muscarinic receptor-PI breakdown relationship in the remaining ovary.

Botulinum toxin A, brain and pain.

Botulinum neurotoxin type A (BoNT/A) is one of the most potent toxins known and a potential biological threat. At the same time, it is among the most widely used therapeutic proteins used yearly by millions of people, especially for cosmetic purposes. Currently, its clinical use in certain types of pain is increasing, and its long-term duration of effects represents a special clinical value. Efficacy of BoNT/A in different types of pain has been found in numerous clinical trials and case reports, as well as in animal pain models. However, sites and mechanisms of BoNT/A actions involved in nociception are a matter of controversy. In analogy with well known neuroparalytic effects in peripheral cholinergic synapses, presently dominant opinion is that BoNT/A exerts pain reduction by inhibiting peripheral neurotransmitter/inflammatory mediator release from sensory nerves. On the other hand, growing number of behavioral and immunohistochemical studies demonstrated the requirement of axonal transport for BoNT/A's antinociceptive action. In addition, toxin's enzymatic activity in central sensory regions was clearly identified after its peripheral application. Apart from general pharmacology, this review summarizes the clinical and experimental evidence for BoNT/A antinociceptive activity and compares the data in favor of peripheral vs. central site and mechanism of action. Based on literature review and published results from our laboratory we propose that the hypothesis of peripheral site of BoNT/A action is not sufficient to explain the experimental data collected up to now.

Botulinum toxin type A selectivity for certain types of pain is associated with capsaicin-sensitive neurons.

Unlike most classical analgesics, botulinum toxin type A (BoNT/A) does not alter acute nociceptive thresholds, and shows selectivity primarily for allodynic and hyperalgesic responses in certain pain conditions. We hypothesized that this phenomenon might be explained by characterizing the sensory neurons targeted by BoNT/A in the central nervous system after its axonal transport. BoNT/A's central antinociceptive activity following its application into the rat whisker pad was examined in trigeminal nucleus caudalis (TNC) and higher-level nociceptive brain areas using BoNT/A-cleaved synaptosomal-associated protein 25 (SNAP-25) and c-Fos immunohistochemistry. Occurrence of cleaved SNAP-25 in TNC was examined after nonselective ganglion ablation with formalin or selective denervation of capsaicin-sensitive (vanilloid receptor-1 or TRPV1-expressing) neurons, and in relation to different cellular and neuronal markers. Regional c-Fos activation and effect of TRPV1-expressing afferent denervation on toxin's antinociceptive action were studied in formalin-induced orofacial pain. BoNT/A-cleaved SNAP-25 was observed in TNC, but not in higher-level nociceptive nuclei. Cleaved SNAP-25 in TNC disappeared after formalin-induced trigeminal ganglion ablation or capsaicin-induced sensory denervation. Occurrence of cleaved SNAP-25 in TNC and BoNT/A antinociceptive activity in formalin-induced orofacial pain were prevented by denervation with capsaicin. Cleaved SNAP-25 localization demonstrated toxin's presynaptic activity in TRPV1-expressing neurons. BoNT/A reduced the c-Fos activation in TNC, locus coeruleus, and periaqueductal gray. Present experiments suggest that BoNT/A alters the nociceptive transmission at the central synapse of primary afferents. Targeting of TRPV1-expressing neurons might be associated with observed selectivity of BoNT/A action only in certain types of pain.