University of Kentucky / University of Pittsburgh Collaborative Center for Analgesia Research Excellence

University of Kentucky / University of Pittsburgh Collaborative Center for Analgesia Research Excellence Project 1: Aim: To determine S1PR1 and astrocyte mechanisms of multiple sclerosis pain. Using the EAE mouse model of multiple sclerosis, drugs will be injected by the subcutaneous or intrathecal route. Pain-like behavior will be assessed with von Frey hairs or using a conditioned place preference assay. Tissues will be collected for immunohistochemical localization of pERK in the spinal cord. These studies will be conducted by Ben Shaw in consultation with Liping Zhang and Bret Smith. Project 2 (Lilian Custodio-Patsey, Carolyn Grachen, Liping Zhang): Aim: To determine spinal and amygdala mechanisms of kappa opioid receptor analgesia. Using the mouse plantar incision model of postoperative pain, EPAC inhibitors will be injected by the intrathecal route. Pain-like behaviors will be assessed with von Frey hairs. Brain tissues will be analyzed for immunohistochemical localization of Fos in the amygdala. Spinal cord tissues will be analyzed by western blot for expression of AMPA receptors. These studies will be conducted by Lilian Custodio-Patsey in consultation with Liping Zhang and Bret Smith. Project 3 (Caitlin Wessel, Romulo Albuquerque): Aim: To determine mechanisms of NPY analgesia. Using the spared nerve injury model of neuropathic pain, NPY receptor inhibitors will be injected by the intrathecal route. Pain-like behaviors will be assessed with our conditioned place preference boxes in Room MS641. These studies will be conducted by Caitlin Wessel in consultation with Liping Zhang. Dr. Albuquerque will assist in the monitoring of this student’s progress. Project 4 Aim: To determine the analgesic efficacy of EPAC inhibitors in a mouse model of orofacial neuropathic pain. EPAC inhibitors will be administered by the intracisternal route. Pain-like behavior will be assessed with von Frey hairs or using a conditioned place preference assay. Tissues will be collected for immunohistochemical localization of pERK in the trigeminal nucleus. These studies will be conducted by Liping Zhang and Robert Danaher. Project 5. Animal breeding. Six mouse lines (AC1-/-, conditional NPYtet-tet, conditional Y1fl/fl, and conditional Y2fl/fl, S1PR1-GFP, and Y1r-Cre) will be maintained until the colonies can be established at the University of Pittsburgh. Breeding and PCR genotyping will be conducted by Tanya Seward. Animal protocols will be overseen by Drs. Danaher and Albuquerque. Reporting and monitoring: Experimental Designs, data spreadsheets, graphs, powerpoint slide sets, and narrative reports will be sent by various personnel to Dr. Taylor on a weekly basis if not more frequently. Dr. Taylor and the various personnel will then discuss the results with further e-mails or Skype calls, or in person at U. Kentucky on a quarterly basis.