The long term objective of this project is to elucidate the role of craniofacial primary afferent neurons in musculoskeletal disorders such as temporomandibular disorders (TMD) and fibromyalgia (FM) using animal models. Two hypotheses are proposed: Hypothesis 1) Masticatory muscle inflammation increases the number of trigeminal ganglion (TG) muscle afferent neurons that express: substance P (SP), calcitonin gene-related peptide (CGRP), neurokinin-1 receptor (NK-lr) and CGRP receptor (CGRPr). This increase involves a phenotypic switch in which muscle primary afferent neurons that do not normally express neuropeptides express SP, CGRP, NK-1r, CGRPr following inflammation. We propose that this change contributes to muscle allodynia and hyperalgesia and can be modulated by pharmacologic manipulations thus providing insight into therapeutics for deep tissue pain. This hypothesis will be tested by quantifying the distribution of TG muscle afferent somata and peripheral axons containing SP, CGRP, NK-1r, CGRPr in three groups: i) control, ii) inflamed muscle, iii) inflamed muscle with intervention (anti-nerve growth factor, NK-lr and CGRPr antagonists). This hypothesis will also be tested by determining the levels ofCGRP, SP and gene expression for CGRP, SP within the TG using radioimmunoassay and reverse transcriptase polymerase chain reaction. Hypothesis 2) SP and CGRP alter the functional properties of TG muscle afferent neurons in part by evoking spontaneous activity and increasing their excitability. We predict that substantially more group II, III and IV TG muscle afferent neurons will be modulated by SP and CGRP following inflammation and that these functional alterations can be modulated pharmacologically. This hypothesis will be tested by characterizing the a) spontaneous and evoked activity and b) active and passive membrane properties of TG muscle afferent neurons prior to muscle inflammation, following muscle inflammation, and following muscle inflammation combined with pharmacological intervention. This will be achieved using intracellular electrophysiological recordings from masseter muscle afferent neurons in a trigeminal ganglion-masseter nerve in vitro preparation. Determination of soma size, axon diameter, and SP, CGRP immunoreactivity for physiologically characterized TG muscle afferent neurons will also test Hypothesis 1. Because a gender difference is reported for TMD and FM, both hypotheses will be tested in males, estrous females and diestrous females.
