title: Upregulation of presynaptic proteins and protein kinases associated with enhanced glutamate release from axonal terminals (synaptosomes) of the medial prefrontal cortex in rats with neuropathic pain.
abstract: Although nerve injury-induced long-term postsynaptic changes have been investigated, less is known regarding the molecular mechanisms within presynaptic axonal terminals. We investigated the molecular changes in presynaptic nerve terminals underlying chronic pain-induced plastic changes in the medial prefrontal cortex (mPFC). After neuropathic pain was induced by spared nerve injury (SNI) in rats, we assessed the release of the excitatory neurotransmitter glutamate by using in vitro synaptosomal preparations from the mPFC. We also measured the levels of synaptic proteins and protein kinases in synaptosomes using Western blotting. The results showed that unilateral long-term SNI augmented depolarization-evoked glutamate release from synaptosomes of the bilateral mPFC. This result was confirmed by a rapid destaining rate of FM1-43 dye in SNI-operated rats. Unilateral long-term nerve injury also significantly increased synaptic proteins (including synaptophysin, synaptotagmin, synaptobrevin, syntaxin, and 25-kDa synaptosome-associated protein) in synaptosomal fractions from the bilateral mPFC, and ultrastructure images demonstrated increased synaptic vesicular profiles in synaptosomes from SNI animals. Chronic pain upregulated the phosphorylation of endogenous protein kinases, including extracellular signal-regulated kinases 1 and 2 (ERK1/2) and Ca(2+)/calmodulin-dependent kinase II (CaMKII), and synapsin I, the primary presynaptic target of ERK1/2 and CaMKII. Both presynaptic proteins and protein kinases were upregulated after SNI in a time-dependent manner. These results indicate that the long-term neuropathic pain-induced enhancement of glutamate release in the mPFC is linked to increased synaptic vesicle proteins and the activation of the ERK1/2- and CaMKII-synapsin signaling cascade in presynaptic axonal terminals.
authors.affiliation: School of Medicine, Fu Jen Catholic University, New Taipei City, Taiwan Department of Pediatrics, Cathay General Hospital, Taipei City, Taiwan Graduate Institute of Basic Medicine, Fu Jen Catholic University, New Taipei City, Taiwan Department of Clinical Psychology, Fu Jen Catholic University, New Taipei City, Taiwan Department of Neurology, Cathay General Hospital, Taipei City, Taiwan Department of Neurology, School of Medicine, National Taiwan University, Taipei City, Taiwan.
authors.forename: Kun-Long
authors.initials: KL
authors.lastname: Hung
authors.forename: Su-Jane
authors.initials: SJ
authors.lastname: Wang
authors.forename: Ying-Chou
authors.initials: YC
authors.lastname: Wang
authors.forename: Tsuey-Ru
authors.initials: TR
authors.lastname: Chiang
authors.forename: Chia-Chuan
authors.initials: CC
authors.lastname: Wang
mesh_terms: D000818:Animals; D018698:Glutamic Acid / Q000378:metabolism*; D008297:Male; D008565:Membrane Proteins / Q000096:biosynthesis*; D009437:Neuralgia / Q000378:metabolism* / Q000473:pathology; D017397:Prefrontal Cortex / Q000378:metabolism*; D017729:Presynaptic Terminals / Q000378:metabolism*; D011494:Protein Kinases / Q000096:biosynthesis*; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D013574:Synaptosomes / Q000378:metabolism; D015854:Up-Regulation / Q000502:physiology
chemical_list: D008565:Membrane Proteins; D018698:Glutamic Acid; D011494:Protein Kinases
keywords: Glutamate; Medial prefrontal cortex; Neuropathic pain; Protein kinase; Synaptic proteins; Synaptosome