Ya-Yun Wang

Crosstalk between Spinal Astrocytes and Neurons in Nerve Injury-Induced Neuropathic Pain

Emerging research implicates the participation of spinal dorsal horn (SDH) neurons and astrocytes in nerve injury-induced neuropathic pain. However, the crosstalk between spinal astrocytes and neurons in neuropathic pain is not clear. Using a lumbar 5 (L5) spinal nerve ligation (SNL) pain model, we testified our hypothesis that SDH neurons and astrocytes reciprocally regulate each other to maintain the persistent neuropathic pain states. Glial fibrillary acidic protein (GFAP) was used as the astrocytic specific marker and Fos, protein of the protooncogene c-fos, was used as a marker for activated neurons. SNL induced a significant mechanical allodynia as well as activated SDH neurons indicated by the Fos expression at the early phase and activated astrocytes with the increased expression of GFAP during the late phase of pain, respectively. Intrathecal administration of c-fos antisense oligodeoxynucleotides (ASO) or astroglial toxin L-α-aminoadipate (L-AA) reversed the mechanical allodynia, respectively. Immunofluorescent histochemistry revealed that intrathecal administration of c-fos ASO significantly suppressed activation of not only neurons but also astrocytes induced by SNL. Meanwhile, L-AA shortened the duration of neuronal activation by SNL. Our data offers evidence that neuronal and astrocytic activations are closely related with the maintenance of neuropathic pain through a reciprocal “crosstalk”. The current study suggests that neuronal and non-neuronal elements should be taken integrally into consideration for nociceptive transmission, and that the intervention of such interaction may offer some novel pain therapeutic strategies.

Changes of the expression of 5-HT receptor subtype mRNAs in rat dorsal root ganglion by complete Freund's adjuvant-induced inflammation.

By using the reverse transcriptase polymerase chain reaction technique, the expression of 5-hydroxytryptamine (5-HT) receptor subtype mRNAs in the rat lumbar dorsal root ganglion (DRG) was investigated following unilateral injection of complete Freunds adjuvant (CFA) into the rat hind paw. The results showed that 5-HT(1A), 5-HT(1B), 5-HT(1D), 5-HT(1F), 5-HT(2A), 5-HT(3), 5-HT(4), 5-HT(5A) and 5-HT(7) receptor subtypes were present in the rat lumbar DRG. CFA injection resulted in a significant increase in mRNA level of 5-HT(1A), 5-HT(1B), 5-HT(1F), 5-HT(2A), 5-HT(3), 5-HT(4) and 5-HT(7) receptor subtypes and a marked induction of 5-HT(2C) subtype mRNA in the DRG. The present results suggest the important roles for these 5-HT receptor subtypes in generating peripheral nociceptive signaling and provide evidence to elucidate the mechanism of 5-HT in nociception.

Changes of 5-HT receptor subtype mRNAs in rat dorsal root ganglion by bee venom-induced inflammatory pain

The reverse transcriptase polymerase chain reaction (RT-PCR) technique was used to examine the changes of the expression of 5-hydroxytryptamine (5-HT) receptors in the rat lumbar dorsal root ganglion (DRG) following subcutaneous bee venom (BV) injection into the plantar surface of the unilateral hindpaw. In the DRG ipsilateral to the BV injection, significant increase of mRNA levels of 5-HT(1A), 5-HT(1B), 5-HT(2A) and 5-HT(3) receptor subtypes were observed at 1 and 4h after the BV injection, while increase of 5-HT(2C), 5-HT(4), 5-HT(6) and 5-HT(7) receptor subtype mRNAs was detected at 4h only. No such changes were observed in the expressions of 5-HT(1D), 5-HT(1F) and 5-HT(5A) receptor subtype mRNAs. Upregulation of 5-HT(1A), 5-HT(1B) and 5-HT(2A) receptor subtype mRNAs was also observed in the contralateral DRG at 4 h. The presence of 5-HT(1E), 5-HT(2B) and 5-HT(5B) receptor subtype mRNAs was not detected in the rat DRG. The present results suggest that different sets of 5-HT receptor subtypes work at different stages of the inflammatory pain induced by subcutaneous BV injection.

Temporal changes of astrocyte activation and glutamate transporter-1 expression in the spinal cord after spinal nerve ligation-induced neuropathic pain.

Astrocyte activation is involved in the neuropathic pain. As a glutamate scavenger, the glutamate transporter‐1 (GLT‐1) is exclusively expressed on the astrocytes and probably correlates with astrocyte activation. In the present study, we attempted to clarify the temporal changing courses of astrocyte activation and GLT‐1 expression, as well as their correlations induced by a neuropathic pain model, namely, spinal nerve ligation (SNL) in which rapidly appearing (<3 days) and persistent (>21 days) mechanical allodynia and thermal hyperalgesia were presented. Immunofluorescent staining showed that GLT‐1 was expressed exclusively in most (not all) of the astrocytes, even when the GLT‐1 expression reached its peak. The expression of GLT‐1 displayed an interesting biphasic change, with an initial up‐regulation followed by a down‐regulation after SNL. Our results also demonstrated that SNL induced a marked and long‐term (>21 days) activation of astrocytes in the ipsilateral spinal dorsal horn. These results suggest that astrocyte activation, the change of GLT‐1 expression and the potential relationship between them might play key roles in the induction and/or maintenance of neuropathic pain. The present results provide novel clues in understanding the mechanisms underlying the involvement of astrocytes and GLT‐1 in the neuropathic pain. Anat Rec, 291:513–518, 2008.

The synaptic connectivity that underlies the noxious transmission and modulation within the superficial dorsal horn of the spinal cord

Noxious stimuli can usually cause the aversive sensations, pain and itch. The initial integration of such noxious information occurs in the superficial dorsal horn of the spinal cord (SDH), which is very important for understanding pain sensation and developing effective analgesic strategies. The circuits formed by pools of neurons and terminals within SDH are accepted as the platform for such complicated integrations and are highly plastic under conditions of inflammatory or neuropathic pain. Recent literature offers a complicated, yet versatile view of SDH intrinsic circuits with both inhibitory and excitatory components. However, our knowledge about the adaptative regulation of SDH local circuits is still far from sufficient due to the incomplete understanding of their organization as they are intermingled with primary afferent fibers (PAFs), poorly understood or identified SDH neurons, somehow contradictory data for descending control systems. A more positive view emphasizes abundant modern data on SDH neuron morphology and physiology riding on the back of significant technological advancements used in neuroscience. Reviewing the current literature on this topic thus produced an integrated understanding of SDH neurons and the SDH local circuits involved in noxious transmission and modulation.

A closer look to botulinum neurotoxin type A-induced analgesia

Chronic pain indicates a type of pain that lasts over time and is accompanied by diagnostic and therapeutic difficulties. It follows that treatment failures are common and patients roam from doctor to doctor in search of an effective care program. So there is an urgent need for long-acting and effective therapeutics to alleviate symptoms of the varied forms of chronic pain. During the past few years, a good success has been achieved with a derivative of a neurotoxin. It has been shown that administration of this toxin can block the release of neurotransmitters and pain mediators. Botulinum neurotoxin type A (BoNT/A) is well known as a treatment for neuromuscular conditions such as dystonia and spasticity. However, the clinical application for BoNT/A has continued to expand. Its analgesic effect has been used in clinical practice with satisfactory results. This review provides an introduction of a hypothesis for the mechanism by which BoNT/A eases chronic pain. It also summarizes the clinical therapeutic effects of BoNT/A in different types of chronic pain and its potential prospects.

5-Hydroxytryptamine1A receptor is involved in the bee venom induced inflammatory pain

&NA; Injection of bee venom into one hindpaw of rat can elicit acute inflammation together with spontaneous pain, heat hyperalgesia and mechanical hyperalgesia/allodynia in the injected paw. 5‐Hydroxytryptamine (5‐HT)1A receptor is the predominant receptor subtype in the spinal dorsal horn mediating the function of 5‐HT in nociception. The goal of the present study is to assess the role of 5‐HT1A receptor in the pain associated with the bee venom induced inflammation. Here we showed that 1 or 4 h after a subcutaneous bee venom challenge, expression of 5‐HT1A receptor mRNA in the ipsilateral lumbar spinal cord increased significantly by 80.94 or 37.86%, respectively. Antisense oligodeoxynucleotide knockdown of spinal 5‐HT1A receptor attenuated spontaneous pain and reversed heat hyperalgesia in rats injected with bee venom. Thus, the present data suggest a facilitating role for 5‐HT1A receptor in bee venom induced inflammatory pain.

Preproenkephalin mRNA is Expressed in a Subpopulation of GABAergic Neurons in the Spinal Dorsal Horn of the GAD67-GFP Knock-In Mouse

GABA (γ‐aminobutyric acid)ergic neurons in the spinal dorsal horn have been reported to be divided into distinctive populations, with different cotransmitters and neuropeptides. In this study, we examined the colocalization of enkephalin (ENK) mRNA with GABA in the spinal dorsal horn using the glutamic acid decarboxylase (GAD)67‐green fluorescence protein (GFP) knock‐in mouse. Our approach was to perform in situ hybridization histochemistry to detect mRNA for preproenkephalin (PPE, the precursor protein for ENK), combined with immunohistochemistry for GFP to reveal GABAergic neurons. Quantitative analysis indicated that more than 44.4% (2967/6681) of GFP‐immunoreactive neurons showed signals for PPE mRNA in the spinal dorsal horn. While 53.9% (2967/5501) of PPE mRNA‐expressing neurons were immunoreactive for GFP. The double‐labeled neurons were observed throughout the spinal dorsal horn, although they had a preferential localization in superficial layers. The present results provide a detailed morphological evidence that ENK and GABA colocalized in a subpopulation of neurons in the spinal dorsal horn, which are likely to represent local inhibitory dorsal horn interneurons involved in the modulation of pain transmission. Anat Rec, 291:1334–1341, 2008.

Understanding the role of mitochondria in the pathogenesis of chronic pain

Chronic pain is a major public health problem. Mitochondria play important roles in a myriad of cellular processes and mitochondrial dysfunction has been implicated in multiple neurological disorders. This review aims to provide an insight into advances in understanding of the role of mitochondrial dysfunction in the pathogenesis of chronic pain. The results show that the five major mitochondrial functions (the mitochondrial energy generating system, reactive oxygen species generation, mitochondrial permeability transition pore, apoptotic pathways and intracellular calcium mobilisation) may play critical roles in neuropathic and inflammatory pain. Therefore, protecting mitochondrial function would be a promising strategy to alleviate or prevent chronic pain states. Related chronic inflammatory and neuropathic pain models, as well as the spectral characteristics of current fluorescent probes to detect mitochondria in pain studies, are also discussed.

The Roles of Neurotensin and its Analogues in Pain

Neurotensin (NT) is an endogenous 13 amino acid neuropeptide with profound opioid-independent analgesic effects. This role of NT is thought to be mediated by both neurotensin receptor subtype 1 (NTS1) and neurotensin receptor subtype 2 (NTS2). NT and its receptors are widely distributed in the pain circuits in central nervous system. Thus NT might modulate pain in many structures of pain pathway, such as spinal cord, rostroventral medulla (RVM) and periaqueductal gray (PAG). Actually either intrathecal application of NT or direct injection of NT into RVM or PAG or intracerebroventricular injection of NT showed analgesic effects. NT exerted its antinociceptive effects in both acute pain and chronic pain models. The analgesic effects of NT were originally found in acute pain experiments. In the case of pathological pain, for example, formalin injection induced inflammatory pain and sciatic nerve constriction induced neuropathic pain, NT also shows antinociceptive effects. The effects exist in somatic pain as well as visceral pain induced by noxious colorectal distension (CRD) or writhing test. It should be noted that NT plays an important role in stress-induced antinociception (SIAN), especially in higher intensity stress experiments. However as a neuropeptide, NT is susceptible to degradation by peptidases and cannot cross the blood-brain barrier (BBB). Great efforts have been made to find NT analogues that are more biologically stable and could inhibit pain by systematic administration. The present review focuses on the analgesic role and the underlying mechanisms of NT and its analogues in pain, especially in chronic pain models.