vasoactive-intestinal-peptide and resiniferatoxin

The present study investigated the influence of intravesically instilled resiniferatoxin (RTX) or tetrodotoxin (TTX) on the distribution, number, and chemical coding of noradrenergic and cholinergic nerve fibers (NF) supplying the urinary bladder in female pigs. Samples from the bladder wall were processed for double-labelling immunofluorescence with antibodies against cholinergic and noradrenergic markers and some other neurotransmitter substances. Both RTX and TTX caused a significant decrease in the number of cholinergic NF in the urinary bladder wall (in the muscle coat, submucosa, and beneath the urothelium). RTX instillation resulted in a decrease in the number of noradrenergic NF in the submucosa and urothelium, while TTX treatment caused a significant increase in the number of these axons in all the layers. The most remarkable changes in the chemical coding of the NF comprised a distinct decrease in the number of the cholinergic NF immunoreactive to CGRP (calcitonin gene-related peptide), nNOS (neuronal nitric oxide synthase), SOM (somatostatin) or VIP (vasoactive intestinal polypeptide), and an increase in the number of noradrenergic NF immunopositive to GAL (galanin) or nNOS, both after RTX or TTX instillation. The present study is the first to suggest that both RTX and TTX can modify the number of noradrenergic and cholinergic NF supplying the porcine urinary bladder.

Topics: Adrenergic Fibers; Animals; Calcitonin Gene-Related Peptide; Cholinergic Fibers; Diterpenes; Female; Galanin; Neuropeptide Y; Nitric Oxide Synthase Type I; Somatostatin; Swine; Tetrodotoxin; Urinary Bladder; Vasoactive Intestinal Peptide; Vesicular Acetylcholine Transport Proteins

In the present study the decrease of neuropeptide containing nerve fibers and the increase in the volume threshold to reflex micturition occurring in the rat bladder after intravesical application of capsaicin or resiniferatoxin were compared. The ultrastructure of bladder terminal axons was evaluated at the moment of maximal peptide depletion and compared to that of nerve fibers after systemic capsaicin application. Adult Wistar rats were treated intravesically for 30 min with 0.5 ml of 100 nM RTX, 1 mM capsaicin or 30% ethanol in saline, the vehicle solution. Twenty-four hours and 1, 2, 3, 4 and 8 weeks later the bladders were immunostained for CGRP, SP, VIP and NPY. Cystomanometric studies were performed 24 h and 1, 8, and 12 weeks after vanilloid instillation. Twenty-four hours after systemic capsaicin or intravesical capsaicin or RTX, bladders were prepared for electron microscopic (EM) observation. Intravesical capsaicin or RTX decreased, in a similar way, the number of CGRP and SP-IR (immunoreactive) fibers coursing in the muscular layer and the mucosa. IR fibers amounted to less than 20% of controls at 24 h and returned to normal levels in the eighth week. At the EM level, bladders treated with topical vanilloids did not show morphological changes in terminal axons coursing in the mucosa. In contrast, bladders from animals treated systemically with capsaicin contained numerous grossly degenerated nerve fibers. VIP and NPY-IR fibers were not affected by the treatment. Cystometrograms showed an increase of the volume threshold to reflex micturition that started at 24 h and disappeared at 12 weeks. We conclude that intravesical capsaicin or RTX were equally effective in terms of reducing the number of SP and CGRP-IR fibers and increasing the volume threshold for reflex micturition. Both changes were transient and were not associated with ultrastructural changes of the bladder nerve fibers, excluding terminal axon degeneration as the main mechanism of action of intravesical vanilloids.

Topics: Animals; Axons; Calcitonin Gene-Related Peptide; Capsaicin; Diterpenes; Female; Male; Microscopy, Electron; Neurons, Afferent; Neuropeptide Y; Neuropeptides; Pressure; Rats; Rats, Wistar; Reflex; Substance P; Urinary Bladder; Urination; Vasoactive Intestinal Peptide

1. Antidromic vasodilatation and plasma extravasation to stimulation of the trigeminal ganglion or its perivascular meningeal fibres was investigated by laser-Doppler flowmetry and 125I-labelled bovin serum albumin in the dura mater and in exteroceptive areas (nasal mucosa, upper eyelid) of anaesthetized rats pretreated with guanethidine and pipecuronium. 2 Trigeminal stimulation at 5 Hz for 20 s elicited unilateral phasic vasodilatation in the dura and lasting response in the nasal mucosa. Resiniferatoxin (1-3 microg kg(-1) i.v.), topical (1%) or systemic capsaicin pretreatment (300 mg kg(-1) s.c. plus 1 mg kg(-1) i.v.) did not inhibit the meningeal responses but abolished or strongly inhibited the nasal responses. Administration of vinpocetine (3 mg kg(-1) i.v.) increased both basal blood flow and the dural vasodilatation to perivascular nerve stimulation. 3. Dural vasodilatation to trigeminal stimulation was not inhibited by the calcitonin gene-related peptide-1 receptor (CGRP-1) antagonist hCGRP8-37 (15 or 50 microg kg(-1) i.v), or the neurokinin-1 receptor antagonist RP 67580 (0.1 mg kg(-1) i.v.) although both antagonists inhibited the nasal response. Neither mucosal nor meningeal responses were inhibited by atropine (5 mg kg(-1) i.v.), hexamethonium (10 mg kg(-1) i.v.) or the vasoactive intestinal polypeptide (VIP) antagonist (p-chloro-D-Phe6-Leul7)VIP (20 microg kg(-1) i.v.). 4. Plasma extravasation in the dura and upper eyelid elicited by electrical stimulation of the trigeminal ganglion was almost completely abolished in rats pretreated with resiniferatoxin (3 microg kg(-1) i.v.). 5. It is concluded that in the rat meningeal vasodilatation evoked by stimulation of trigeminal fibres is mediated by capsaicin-insensitive primary afferents, while plasma extravasation in the dura and upper eyelid and the vasodilatation in the nasal mucosa are mediated by capsaicin-sensitive trigeminal fibres.

Topics: Animals; Calcitonin Gene-Related Peptide; Calcitonin Gene-Related Peptide Receptor Antagonists; Capsaicin; Diterpenes; Electric Stimulation; Indoles; Isoindoles; Laser-Doppler Flowmetry; Male; Meninges; Nasal Mucosa; Nerve Fibers; Neurokinin-1 Receptor Antagonists; Neurons, Afferent; Neurotoxins; Peptide Fragments; Rats; Rats, Wistar; Regional Blood Flow; Serum Albumin, Radio-Iodinated; Trigeminal Nerve; Vasoactive Intestinal Peptide; Vasodilation

Resiniferatoxin (RTX) depletes vanilloid (capsaicin) receptors from lumbar dorsal root ganglia (DRG) of the rat. In addition, RTX causes changes in neuropeptide and nitric oxide synthase expression in lumbar DRG neurons, similar to those described following axotomy; this latter phenomenon is referred to as messenger plasticity. These findings suggested that vanilloid receptor loss may be part of the plasticity that follows RTX treatment. Here we show that vanilloid receptor expression, as detected by [3H]RTX autoradiography, is not changed in lumbar DRGs of axotomized rats, nor is it altered in a rat model (chronic constriction injury) of neuropathic pain. Thus, the in vivo expression of vanilloid receptors detected by specific [3H]RTX binding does not require the presence of intraaxonally transported trophic factors such as nerve growth factor. We conclude that messenger plasticity and vanilloid receptor loss are mediated by distinct mechanisms.

Topics: Animals; Autoradiography; Capsaicin; Diterpenes; Female; Galanin; Ganglia, Spinal; Lumbosacral Region; Neuronal Plasticity; Neurotoxins; Nitric Oxide Synthase; Rats; Rats, Sprague-Dawley; Receptors, Drug; Vasoactive Intestinal Peptide

Using in situ hybridization, the expression of mRNA encoding galanin, vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY), and nitric oxide synthase (NOS), respectively, was studied in lumbar dorsal root ganglia of rats given a single s.c. dose of 300 micrograms kg-1 resiniferatoxin (RTX), an ultrapotent capsaicin analogue. In control animals, 10% of the DRG neurones were positive for galanin mRNA, whereas no message for VIP, NPY or NOS could be detected. One week after RTX treatment, a markedly increased number (approximately 30%) of the neurones expressed galanin mRNA. Simultaneously, VIP and NOS mRNA became detectable in 6-8% of the neurones. The number of galanin-positive neurones declined after 2 weeks and returned to control levels by 8 weeks. The increase in number of VIP-, or NOS-positive neurones persisted up to 4 weeks after RTX treatment and declined thereafter. Also, there was a small increase in NPY mRNA-positive neurones. In parallel immunohistochemical experiments, similar increases were observed for galanin message-associated protein (GMAP)-, VIP- and NOS-like immunoreactivities. Our findings suggest that RTX can cause changes (messenger plasticity) in galanin, VIP and NOS expression in capsaicin-sensitive sensory neurones of the rat, similar to those described following axotomy.

Topics: Animals; Diterpenes; Female; Galanin; Ganglia, Spinal; Immunohistochemistry; Lumbosacral Region; Neurotoxins; Nitric Oxide Synthase; Peptide Fragments; Rats; Rats, Sprague-Dawley; Vasoactive Intestinal Peptide

Neurochemical and functional studies were performed to investigate and to compare the effects of resiniferatoxin and capsaicin in the rat stomach. Neonatal administration of resiniferatoxin (0.6-1.6 mumol/kg subcutaneously (s.c.)) produced a marked decrease in gastric calcitonin gene-related peptide-like immunoreactivity in both secretory and non-secretory region of the stomach. Almost complete depletion of the peptide was determined by neonatal administration of capsaicin (164 mumol/kg s.c.). Vasoactive intestinal polypeptide-like immunoreactivity was concomitantly unaffected by resiniferatoxin or capsaicin, thus showing the selectivity of action of the neurotoxins on gastric afferent fibers. Oral administration of an equimolar dose (0.3 nmol/kg) of resiniferatoxin or capsaicin together with 50% ethanol reduced at a similar extent gastric haemorrhagic lesions produced by the mucosal barrier-breaker agent. These findings provide evidence that resiniferatoxin and capsaicin may act on a common neuronal target in the rat stomach and that the acute exciting (protective) effect is of the same magnitude.

Topics: Administration, Oral; Animals; Animals, Newborn; Calcitonin Gene-Related Peptide; Capsaicin; Diterpenes; Dose-Response Relationship, Drug; Ethanol; Gastric Mucosa; Gastrointestinal Hemorrhage; Injections, Subcutaneous; Male; Neurons, Afferent; Neurotoxins; Rats; Stomach Ulcer; Vasoactive Intestinal Peptide