Regulatory mechanisms for subtance P in the dorsal horn during a nociceptive stimulus: Axoplasmic transport vs electrical activity

doi:10.1016/0006-8993(86)91074-7

Brain Research

Volume 385, Issue 2, 22 October 1986, Pages 282-290

https://doi.org/10.1016/0006-8993(86)91074-7 Get rights and content

Abstract

Substance P (SP) is believed to be a neuromediator of nociception in the dorsal horn of the spinal cord. SP prescursor is synthesized in the dorsal root ganglia (DRG) and transported via axoplasmic transport to the nerve terminal where it is stored and released as SP. The chemical nociceptive stimulus, formalin, when injected into the hindpaw causes an increase in the level of SP in the dorsal horn. This increase in SP may be the result of increased electrical activity due to activation of free nerve endings or the transport of some chemical or trophic signal to the DRG or to the central terminal. This study investigates the mechanism of the SP increase during the formalin stimulus. Rats were anesthetized and a laminectomy performed. In some experiments the sciatic nerve was exposed. Agar gel pads containing either colchicine or tetrodotoxin (TTX) were applied to the root or sciatic nerve prior injection of 5% formalin or saline into the hindpaw. Electrical activity across the dorsal root distal to the gel pad was monitored to determine the effects of colchicine and TTX on the nerve. Sixty min after the injection into the hindpaw, the animal was perfused and the lumbar spinal cord removed. Ten-μm frozen sections were stained for SP. It was found that the formalin-evoked increase in SP could be partially blocked by either colchicine or TTX applied to the dorsal root and completely blocked by the application of both agents together. TTX or colchicine applied to the sciatic nerve completely blocked the formalin-evoked increase in SP. These data show that the regulation of SP level during a chemical nociceptive stimulus is dependent upon both electrical activity and the axoplasmic transport of some chemical signal.

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This finding is in agreement with studies that employed the formalin test in the rats’ paw, which have shown that pharmacological blockade of NK1 receptors reduced the first or both phases of nociception evoked by formalin (Birch et al., 1992; Traub, 1996). Furthermore, there are several evidences that injection of formalin in the paw leads to an increase of SP levels in the dorsal horn, probably, as the result of an increased biosynthesis, transport and release of this peptide in primary afferents and spinal neurons in response to long-lasting inflow of noxious messages (Kantner et al., 1986; Zhang et al., 1994). Altogether, these results reinforce the role of SP in persistent pain by acting at both, the peripheral level (present study), as well as at the central level, where most of the studies demonstrated increased expression of SP after formalin injection.
There is accumulating evidence that substance P released from peripheral sensory neurons participates in inflammatory and neuropathic pain. In this study it was investigated the ability of substance P to induce orofacial nociception and thermal and mechanical hyperalgesia, as well as the role of NK₁ receptors on models of orofacial inflammatory and neuropathic pain. Substance P injected into the upper lip at 1, 10 and 100 μg/50 μL failed to induce nociceptive behavior. Also, substance P (0.1–10 μg/50 μL) injected into the upper lip did not evoke orofacial cold hyperalgesia and when injected at 1 μg/50 μL did not induce mechanical hyperalgesia. However, substance P at this latter dose induced orofacial heat hyperalgesia, which was reduced by the pre-treatment of rats with a non-peptide NK₁ receptor antagonist (SR140333B, 3 mg/kg). Systemic treatment with SR140333B (3 mg/kg) also reduced carrageenan-induced heat hyperalgesia, but did not exert any influence on carrageenan-induced cold hyperalgesia. Blockade of NK₁ receptors with SR140333B also reduced by about 50% both phases of the formalin response evaluated in the orofacial region. Moreover, heat, but not cold or mechanical, hyperalgesia induced by constriction of the infraorbital nerve, a model of trigeminal neuropathic pain, was abolished by pretreatment with SR140333B. Considering that substance P was peripherally injected (i.e. upper lip) and the NK₁ antagonist used lacks the ability to cross the blood–brain-barrier, our results demonstrate that the peripheral SP/NK₁ system participates in the heat hyperalgesia associated with inflammation or nerve injury and in the persistent pain evoked by formalin in the orofacial region.
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The gel was then cut into 5 mm long and 2 mm wide strips. The strips were placed on the median nerve in a “sandwich” dressing [30]. Group 4 was anesthetized and the right median nerve was exposed.
In this study, we examined the relationship between astrocyte activation in the cuneate nucleus (CN) and behavioral hypersensitivity after chronic constriction injury (CCI) of the median nerve. In addition, we also examined the effects of pre-emptive treatment with a number of drugs on astrocyte activation and hypersensitivity development in this model. Using immunohistochemistry and immunoblotting, little glial fibrillary acidic protein (GFAP; an astrocyte marker) immunoreactivity was detected in the CN of the normal rats. As early as 3 days after CCI, there was a significant increase in GFAP immunoreactivity in the lesion side of CN, and this reached a maximum at 7 days, and was followed by a decline. Counting of GFAP-immunoreactive astrocytes revealed that astrocytic hypertrophy, but not proliferation, contributes to increased GFAP immunoreactivity. Furthermore, microinjection of the glial activation inhibitor, fluorocitrate, into the CN at 3 days after CCI attenuated injury-induced behavioral hypersensitivity in a dose-dependent manner. These results suggest that median nerve injury-induced astrocytic activation in the CN modulated the development of behavioral hypersensitivity. Animals received MK-801 (glutamate N-methyl-d-aspartate (NMDA) receptor antagonist), clonidine (α₂-adrenoreceptor agonist), tetrodotoxin (TTX, sodium channel blocker) or lidocaine (local anesthetic) 30 min prior to median nerve CCI. Pre-treatment with MK-801, TTX, and 2% lidocaine, but not clonidine, attenuated GFAP immunoreactivity and behavioral hypersensitivity following median nerve injury. In conclusion, suppressing reactions to injury, such as the generation of ectopic discharges and activation of NMDA receptors, can decrease astrocyte activation in the CN and attenuate neuropathic pain sensations.
Activation of mu, delta or kappa opioid receptors by DAMGO, DPDPE, U-50488 or U-69593 respectively causes antinociception in the formalin test in the naked mole-rat (Heterocephalus glaber)
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Data available on the role of the opioid systems of the naked mole-rat in nociception is scanty and unique compared to that of other rodents. In the current study, the effect of DAMGO, DPDPE and U-50488 and U-69593 on formalin-induced (20 μl, 10%) nociception were investigated. Nociceptive-like behaviors were quantified by scoring in blocks of 5 min the total amount of time (s) the animal spent scratching/biting the injected paw in the early (0–5 min) and in the late (25–60 min) phase of the test. In both the early and late phases, administration of 1 or 5 mg/kg of DAMGO or DPDPE caused a naloxone-attenuated decrease in the mean scratching/biting time. U-50488 and U-69593 at all the doses tested did not significantly change the mean scratching/biting time in the early phase. However, in the late phase U-50488 or U-69593 at the highest doses tested (1 or 5 mg/kg or 0.025 or 0.05 mg/kg, respectively) caused a statistically significant and naloxone-attenuated decrease in the mean scratching/biting time. The data showed that mu, delta or kappa-selective opioids causes antinociception in the formalin test in this rodent, adding novel information on the role of opioid systems of the animal on pain regulation.
Spinal subarachnoid adrenal medullary transplants reduce hind paw swelling and peripheral nerve transport following formalin injection in rats
2008, Brain Research
Citation Excerpt :
In none of the treatment groups was SP IR noted distally to the ligation, thus confirming anterograde-only transport of SP.) However, formalin injection significantly increased SP IR in sham- and control-transplanted rats. A previous study demonstrated that retarding microtubule function with colchicine treatment of the central terminal significantly suppressed formalin-induced accumulation of SP IR in the dorsal horn (Kantner et al., 1986). Thus, the current data demonstrate that formalin increases axoplasmic transport of SP to the periphery.
Previous studies have demonstrated that adrenal medullary chromaffin cells transplanted into the spinal subarachnoid space significantly reduced pain-related behavior following hind paw plantar formalin injection in rats. The data suggests a centrally mediated antinociceptive mechanism. The spinal transplants may have effects on sciatic nerve function as well. To address this, the current study examined the effects of spinal adrenal transplants on hind paw edema and the anterograde transport of substance P (SP) that occur following formalin injection. Robust formalin-evoked edema, as well as hind paw flinching, was observed in striated muscle control-transplanted rats, which were not observed in adrenal-transplanted rats. To visualize transport of SP, the sciatic nerve was ligated ipsilateral to formalin injection and the nerve was processed 48 h later for immunocytochemistry. A significant formalin-induced accumulation of SP immunoreactivity (IR) was observed proximal to the ligation in control-transplanted rats. In contrast, there was significantly less SP IR observed from nerve of adrenal-transplanted rats, suggesting a diminution of anterograde axoplasmic transport by adrenal transplants. The change in SP IR may have been due to an alteration of transport due to formalin injection, thus, transport was visualized by the accumulation of growth-associated protein 43 (GAP43) at the ligation site. Formalin injection did not significantly increase proximal accumulation of GAP43 IR, indicating that formalin does not increase anterograde transport. Surprisingly, however, adrenal transplants significantly diminished GAP43 IR accumulation compared to control-transplanted rats. These data demonstrate that spinal adrenal transplants can attenuate the formalin-evoked response by modulating primary afferent responses.
Modulation of the formalin-induced nociceptive response by diabetes: Possible involvement of intracellular calcium
2000, Brain Research
We examined the involvement of cytosolic calcium in the modulation of the formalin-induced nociceptive response by diabetes. Injection of formalin into the hindpaw of mice produced a biphasic nociceptive response consisting of immediate (first phase) and tonic (second phase) components. Although the duration of the first-phase response was significantly longer in diabetic mice than in non-diabetic mice, the second phase was significantly shorter in diabetic mice. The first-phase response was dose-dependently and significantly reduced by pretreatment with ryanodine, which blocks Ca²⁺ release from Ca²⁺/caffeine-sensitive microsomal pools. The second-phase response was also significantly increased when diabetic mice were pretreated with ryanodine. However, ryanodine had no significant effect on either the first-phase or second-phase response in non-diabetic mice. On the other hand, pretreatment with thapsigargin, which inhibits Ca²⁺ uptake into the inositol-1,4,5-trisphosphate-sensitive microsomal Ca²⁺ pool, significantly enhanced the first-phase response in non-diabetic mice. Furthermore, thapsigargin significantly and dose-dependently reduced the second phase of the formalin-induced nociceptive response in non-diabetic mice. Thapsigargin administered i.t. did not significantly affect either the first- or the second-phase response in diabetic mice. These results suggest that the change in the formalin-induced nociceptive response in diabetic mice may be due, at least in part, to the modification of nociceptive transmission in the spinal cord by intracellular calcium.
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^∗: Present address: Department of Physical Therapy, Medical College of Ohio, C.S. 10008, 3000 Arlington Avenue, Toledo, OH 43699, U.S.A.

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Research reportRegulatory mechanisms for subtance P in the dorsal horn during a nociceptive stimulus: Axoplasmic transport vs electrical activity

Abstract

Brain Research

Prog. Neurobiol.

Brain Research

Brain Research

Brain Research

Gen. Pharm.

Brain Research

Brain Research

Exp. Neurol.

Brain Research

Brain Research

TINS

Exp. Neurol.

Neuroscience

Neurosci. Lett.

The origin, distribution and synaptic relationships of substance P axons in rat spinal cord

J. Comp. Neurol.

Organization of the Spinal Cord

Effects of lidocaine on axonal morphology, microtubules, and rapid transport in rabbit vagus nerve in vitro

J. Neurobiol.

Transganglionic degenerative atrophy and regenerative proliferation in the Rolando substance of the primate spinal cord: discoupling and restoration of synaptic connectivity in the central nervous system after peripheral nerve lesion

Folia Morphol.

Research report
Regulatory mechanisms for subtance P in the dorsal horn during a nociceptive stimulus: Axoplasmic transport vs electrical activity