Date of Award
Summer 7-28-2017
Degree Type
Thesis
Degree Name
Master of Science in Integrative Biology (MSIB)
Department
Biology
Committee Chair/First Advisor
Dr. Scott Nowak
Major Professor
Dr. Lisa Ganser
Second Committee Member
Dr. Martin Hudson
Abstract
Hyperalgesia is the increased sensitivity to pain which can present itself to many people no matter age or ethnicity. Having the ability to alleviate pain whether it be acute or chronic pain without the use of addictive medication can have the potential to change the course or modern medicine known today. Through the use of zebrafish, this thesis provides a preliminary understanding in pain signal blocking. Three distinct genes have found to be associated with perceiving pain, which include Transient Receptor Potential Ankyrin 1 (TRPA1), Transient Receptor Potential Melestatin 8 (TRPM8), and Cannabinoid Receptor 1 (CNR1). We hypothesize through knockdown of TRPA1, TRPM8, or CNR1 gene function may alleviate chronic pain-based behaviors and pain signaling. Gene manipulation by way of splice site-blocking morpholinos was performed on the stated genes of interest and action potentials were generated through exposure to the noxious stimulus cinnamon oil. Behavioral responses to pain were examined by looking at the characteristic escape response of the morphant and control fish. In addition, electrophysiology was performed on cranial nerve VIII (CNVIII) of morphants to understand whether pain signaling, in response to the noxious, exogenous compound cinnamon oil, persisted even after knockdown of these pain perception genes. The impact of TRP channel knockdown on pain signaling was then observed on the post synaptic level by comparing the fluorescent intensity mean (FIM) values of fluorescent antibody labeled NMDA receptors for glutamate and PSD95 clustering proteins for NMDAr at integration and motor synapses of spinal cord tissues. The FIM values were then correlated to the presence of NMDA, or PSD95 on spinal neurons to indicate production and assembly of glutamatergic synapse components, eluding to colocalization of NMDA receptor and its clustering proteins, even in the absence of a signal.