ABSTRACT
Activation of astrocytes has a profound effect on brain plasticity and is critical for the pathophysiology of several neurological disorders including neuropathic pain. Here, we show that metabotropic glutamate receptor 5 (mGluR5), which reemerges in astrocytes in a restricted time frame, is essential for these functions. Although mGluR5 is absent in healthy adult astrocytes, it transiently reemerges in astrocytes of the somatosensory cortex (S1). During a limited spatiotemporal time frame, astrocytic mGluR5 drives Ca2+ signals; upregulates multiple synaptogenic molecules such as Thrombospondin-1, Glypican-4, and Hevin; causes excess excitatory synaptogenesis; and produces persistent alteration of S1 neuronal activity, leading to mechanical allodynia. All of these events were abolished by the astrocyte-specific deletion of mGluR5. Astrocytes dynamically control synaptic plasticity by turning on and off a single molecule, mGluR5, which defines subsequent persistent brain functions, especially under pathological conditions.
Subject(s)
Astrocytes , Chronic Pain , Animals , Astrocytes/metabolism , Chronic Pain/pathology , Mice , Neuronal Plasticity , Neurons/metabolism , Receptor, Metabotropic Glutamate 5/metabolismABSTRACT
The anticancer agents platinum derivatives and taxanes such as paclitaxel (PCX) often cause neuropathy known as chemotherapy-induced peripheral neuropathy with high frequency. However, the cellular and molecular mechanisms underlying such neuropathy largely remain unknown. Here, we show new findings that the effect of Goshajinkigan (GJG), a Japanese KAMPO medicine, inhibits PCX-induced neuropathy by acting on astrocytes. The administration of PCX in mice caused the sustained neuropathy lasting at least 4 weeks, which included mechanical allodynia and thermal hyperalgesia but not cold allodynia. PCX-evoked pain behaviors were associated with the sensitization of all primary afferent fibers. PCX did not activate microglia or astrocytes in the spinal cord. However, it significantly activated astrocytes in the primary sensory (S1) cortex without affecting S1 microglial activation there. GJG significantly inhibited the PCX-induced mechanical allodynia by 50% and thermal hyperalgesia by 90%, which was in accordance with the abolishment of astrocytic activation in the S1 cortex. Finally, the inhibition of S1 astrocytes by an astrocyte-toxin L-alpha-aminoadipic acid abolished the PCX-induced neuropathy. Our findings suggest that astrocytes in the S1 cortex would play an important role in the pathogenesis of PCX-induced neuropathy and are a potential target for its treatment.
Subject(s)
Drugs, Chinese Herbal/pharmacology , Neuralgia/drug therapy , Paclitaxel/adverse effects , Animals , Antineoplastic Agents, Phytogenic/administration & dosage , Antineoplastic Agents, Phytogenic/adverse effects , Astrocytes/drug effects , Astrocytes/metabolism , Disease Models, Animal , Humans , Hyperalgesia/chemically induced , Hyperalgesia/drug therapy , Mice , Neuralgia/chemically induced , Paclitaxel/administration & dosage , Somatosensory Cortex/cytology , Somatosensory Cortex/drug effectsABSTRACT
Methylated DNA plays an important role in epigenetic gene regulation, and could therefore serve as a potentially promising biomarker for the diagnosis of cancer and other diseases. Therefore, the development of a simple method for analyzing cytosine methylation in a target gene is required. Here, we report on a new method for the sequence-selective chemical modification of a single cytosine in single-stranded DNA (ssDNA). This method is based on the formation of a DNA three-way junction of the ssDNA and two ssDNA probes, and was successfully applied for a simple polymerase chain reaction (PCR)-based assay for 'pinpoint' methylation analysis.
Subject(s)
Cytosine/analysis , Cytosine/metabolism , DNA Probes/chemistry , DNA, Single-Stranded/analysis , DNA Methylation , DNA, Single-Stranded/genetics , DNA, Single-Stranded/metabolismABSTRACT
DNA methylation is an epigenetic mechanism for transcriptional regulation. The methylation process controls cellular differentiation and is defective in many diseases including cancer. Therefore, the development of a simple method for analysing cytosine methylation in a target gene is required. Here we report a conceptually new method for sequence-selective chemical modification of a single cytosine in single-stranded DNA (ssDNA) using two DNA probes to form a DNA three-way junction with the ssDNA. The method was successfully used in a simple quantitative polymerase-chain-reaction-based assay for discrimination of a single methylated cytosine.
