Changes in expression of sensory organ-specific microRNAs in rat dorsal root ganglia in association with mechanical hypersensitivity induced by spinal nerve ligation

Department of Anesthesia, University of Iowa, 51 Newton Road, Iowa City, IA 52242, USA.
Neuroscience (Impact Factor: 3.36). 09/2009; 164(2):711-23. DOI: 10.1016/j.neuroscience.2009.08.033
Source: PubMed


Chronic neuropathic pain caused by peripheral nerve injury is associated with global changes in gene expression in damaged neurons. To understand the molecular mechanisms underlying neuropathic pain, it is essential to elucidate how nerve injury alters gene expression and how the change contributes to the development and maintenance of chronic pain. MicroRNAs are non-protein-coding RNA molecules that regulate gene expression in a wide variety of biological processes mainly at the level of translation. This study investigated the possible involvement of microRNAs in gene regulation relevant to neuropathic pain. The analyses focused on a sensory organ-specific cluster of microRNAs that includes miR-96, -182, and -183. Quantitative real-time polymerase chain reaction (qPCR) analyses confirmed that these microRNAs were highly enriched in the dorsal root ganglion (DRG) of adult rats. Using the L5 spinal nerve ligation (SNL) model of chronic neuropathic pain, we observed a significant reduction in expression of these microRNAs in injured DRG neurons compared to controls. In situ hybridization and immunohistochemical analyses revealed that these microRNAs are expressed in both myelinated (N52 positive) and unmyelinated (IB4 positive) primary afferent neurons. They also revealed that the intracellular distributions of the microRNAs in DRG neurons were dramatically altered in animals with mechanical hypersensitivity. Whereas microRNAs were uniformly distributed within the DRG soma of non-allodynic animals, they were preferentially localized to the periphery of neurons in allodynic animals. The redistribution of microRNAs was associated with changes in the distribution of the stress granule (SG) protein, T-cell intracellular antigen 1 (TIA-1). These data demonstrate that SNL induces changes in expression levels and patterns of miR-96, -182, and -183, implying their possible contribution to chronic neuropathic pain through translational regulation of pain-relevant genes. Moreover, SGs were suggested to be assembled and associated with microRNAs after SNL, which may play a role in modification of microRNA-mediated gene regulation in DRG neurons.

Download full-text


Available from: Donna Hammond
  • Source
    • "This capability is inherent to microRNAs (miRNAs), small non-protein-coding RNAs known to play an important role in development and disease by post-transcriptional down-regulation of entire sets of target mRNAs [9-11]. Evidence supporting this hypothesis includes the facts that miRNAs are up- and down-regulated in sensory neurons in animal models of neuropathy, that certain salient miRNAs are prominently expressed in nociceptors and that pain is suppressed in transgenic mice in which global miRNA expression in nociceptors was selectively blocked [12-20]. This evidence, however, is not specific to pain. "
    [Show abstract] [Hide abstract]
    ABSTRACT: We carried out a genome-wide study, using microRNA sequencing (miRNA-seq), aimed at identifying miRNAs in primary sensory neurons that are associated with neuropathic pain. Such scans usually yield long lists of transcripts regulated by nerve injury, but not necessarily related to pain. To overcome this we tried a novel search strategy: identification of transcripts regulated differentially by nerve injury in rat lines very similar except for a contrasting pain phenotype. Dorsal root ganglia (DRGs) L4 and 5 in the two lines were excised 3 days after spinal nerve ligation surgery (SNL) and small RNAs were extracted and sequenced. We identified 284 mature miRNA species expressed in rat DRGs, including several not previously reported, and 3340 unique small RNA sequences. Baseline expression of miRNA was nearly identical in the two rat lines, consistent with their shared genetic background. In both lines many miRNAs were nominally up- or down-regulated following SNL, but the change was similar across lines. Only 3 miRNAs that were expressed abundantly (rno-miR-30d-5p, rno-miR-125b-5p) or at moderate levels (rno-miR-379-5p) were differentially regulated. This makes them prime candidates as novel PNS determinants of neuropathic pain. The first two are known miRNA regulators of the expression of Tnf, Bdnf and Stat3, gene products intimately associated with neuropathic pain phenotype. A few non-miRNA, small noncoding RNAs (sncRNAs) were also differentially regulated. Despite its genome-wide coverage, our search strategy yielded a remarkably short list of neuropathic pain-related miRNAs. As 2 of the 3 are validated regulators of important pro-nociceptive compounds, it is likely that they contribute to the orchestration of gene expression changes that determine individual variability in pain phenotype. Further research is required to determine whether some of the other known or predicted gene targets of these miRNAs, or of the differentially regulated non-miRNA sncRNAs, also contribute.
    Full-text · Article · Mar 2014 · Molecular Pain
  • Source
    • "Pain conditions have been suggested to deregulate the expression of miRNAs in pain pathways from primary afferent nociceptors to brain areas associated with emotional components of pain perception (Bai et al., 2007; Aldrich et al., 2009; Kusuda et al., 2011; Imai et al., 2011; Poh et al., 2011; von Schack et al., 2011). miRNAs are frequently deregulated and expressed at aberrant levels in diseased tissue, and first evidence suggests that this applies to neurogenic pain in CRPS (Orlova et al., 2011). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Neuro-immune alterations in the peripheral and central nervous system play a role in the pathophysiology of chronic pain, and non-coding RNAs - and microRNAs (miRNAs) in particular - regulate both immune and neuronal processes. Specifically, miRNAs control macromolecular complexes in neurons, glia and immune cells and regulate signals used for neuro-immune communication in the pain pathway. Therefore, miRNAs may be hypothesized as critically important master switches modulating chronic pain. In particular, understanding the concerted function of miRNA in the regulation of nociception and endogenous analgesia and defining the importance of miRNAs in the circuitries and cognitive, emotional and behavioral components involved in pain is expected to shed new light on the enigmatic pathophysiology of neuropathic pain, migraine and complex regional pain syndrome. Specific miRNAs may evolve as new druggable molecular targets for pain prevention and relief. Furthermore, predisposing miRNA expression patterns and inter-individual variations and polymorphisms in miRNAs and/or their binding sites may serve as biomarkers for pain and help to predict individual risks for certain types of pain and responsiveness to analgesic drugs. miRNA-based diagnostics are expected to develop into hands-on tools that allow better patient stratification, improved mechanism-based treatment, and targeted prevention strategies for high risk individuals.
    Full-text · Article · Oct 2013 · Frontiers in Molecular Neuroscience
  • Source
    • "Recent studies showed that miRNAs are abundantly expressed in the nervous system and have been initially identified as critical mediators in the regulation of neuroinflammation and neuropathic pain (Imai et al., 2011; Kosik, 2006; Tarassishin et al., 2011). Aldrich et al. (2009) found that miR-96, miR-182, and miR-183 are highly enriched in the dorsal root ganglion of adult rats, and spinal nerve ligation (SNL) results in a significant reduction in expression of these miRNAs. Further research implied their possible contribution to chronic neuropathic pain through translational regulation of pain-relevant genes. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Unlabelled: Following peripheral nerve injury (PNI) microglia proliferates and adopts inflammation that contributes to development and maintenance of neuropathic pain. miRNAs and autophagy are two important factors in the regulation of inflammation. However, little is known about whether miRNAs regulate neuroinflammation and neuropathic pain by controlling autophagy. In the study, we demonstrated that miR-195 levels were markedly increased in rats subjected to L5 spinal nerve ligation (SNL). Upregulated miR-195 was also found in spinal microglia of rats with SNL. The overexpression of miR-195 contributed to lipopolysaccharide-induced expression of proinflammatory cytokines IL-1β, TNF-α, and iNOS in cultured microglia. Upregulated miR-195 also resulted in increased mechanical and cold hypersensitivity after PNI, whereas miR-195 inhibition reduced mechanical and cold sensitivity. We further demonstrated that PNI significantly inhibited microglial autophagy activation, whereas miR-195 inhibitor treatment increased autophagy activation and suppressed neuroinflammation in vivo and in vitro. More important, autophagy inhibition impaired miR-195 inhibitor-induced downregulation of neuroinflammation and neuropathic pain. Additionally, ATG14 was identified as the functional target of miR-195. Conclusions: These data demonstrated that miR-195/autophagy signaling represents a novel pathway regulating neuroinflammation and neuropathic pain, thus offering a new target for therapy of neuropathic pain.
    Full-text · Article · Apr 2013 · Glia
Show more