title: Contribution of Primary Afferent Input to Trigeminal Astroglial Hyperactivity, Cytokine Induction and NMDA Receptor Phosphorylation.
abstract: We tested the hypothesis that primary afferent inputs play a role in astroglial hyperactivity after tissue injury. We first injected complete Freund's adjuvant (CFA, 0.05 ml, 1:1 oil/saline) into the masseter muscle, which upregulated glial fibrillary acidic protein (GFAP), a marker of astrocytes, interleukin (IL)-1β an inflammatory cytokine, and phosphorylation of serine896 of the NR1 subunit (P-NR1) of the NMDA receptor in the subnuclei interpolaris/caudalis (Vi/Vc) transition zone, an important structure for processing trigeminal nociceptive input. Local anesthetic block with lidocaine (2%) of the masseter muscle at 10 min prior to injection of CFA into the same site significantly reduced the CFA-induced increase in GFAP, IL-1β and P-NR1 (p<0.05, n=4/group). We then tested the effect of peripheral electrical stimulation (ES). The ES protocol was burst stimulation consisting of trains of 4 square pulses (10-100 Hz, 0.1-3 mA, 0.5 ms pulse width). Under pentobarbital anesthesia, an ES was delivered every 0.2 s for a total of 30 min. The Vi/Vc tissues were processed for immunohistochemistry or western blot analysis at 10-120 min after ES. Compared to naive and SHAM-treated rats, there was increased immunoreactivity against GFAP, IL-1β and P-NR1 in the Vi/Vc in rats receiving ES. Double staining showed that IL-1β was selectively localized in GFAP-positive astroglia, and P-NR1-immunoreactivity was localized to neurons. These findings indicate that primary afferent inputs are necessary and sufficient to induce astroglial hyperactivity and upregulation of IL-1β, as well as neuronal NMDA receptor phosphorylation.
authors.affiliation: Department of Neural and Pain Sciences, Dental School and Program in Neuroscience, University of Maryland, Baltimore, MD 21201, USA.
authors.forename: H
authors.initials: H
authors.lastname: Wang
authors.forename: W
authors.initials: W
authors.lastname: Guo
authors.forename: K
authors.initials: K
authors.lastname: Yang
authors.forename: F
authors.initials: F
authors.lastname: Wei
authors.forename: R
authors.initials: R
authors.lastname: Dubner
authors.forename: K
authors.initials: K
authors.lastname: Ren