First Advisor
Aaron K. Apawu
First Committee Member
Murielle Watzky
Second Committee Member
Corina Brown
Third Committee Member
Patrick Burns
Degree Name
Master of Science
Document Type
Thesis
Date Created
8-2025
Department
College of Natural and Health Sciences, Chemistry and Biochemistry, Chemistry and Biochemistry Student Work
Embargo Date
8-2026
Abstract
Neuropeptides are short sequences of amino acids that function along with classical neurotransmitters in the central nervous system. They serve as neuromodulators in the neuronal circuits by affecting the synaptic transmission and serve as signaling molecules that induce intracellular response by activating many intercellular signaling pathways. Due to their extensive roles, they are often implicated in several neurological disorders as well as hearing related disorders. Traditionally, neuropeptides were studied using the common immunoassay methods such as ELISA, Edman Sequencing, and Western blotting. However, these traditional methods have limitations in terms of robustness, the detection of post-translational modifications, and prior knowledge of the peptide sequence is required. Alternatively, liquid chromatography-tandem mass spectrometry (LC-MS/MS) has revolutionized the field of peptidomics, as it allows the analysis of neuropeptides that bear post-translational modifications, and requires minimal sample volumes, and no need for prior knowledge of a particular peptide sequence. Although mass spectrometry based peptidomics is a robust and effective method for neuropeptide analysis, the extraction of neuropeptides may pose some challenges in terms of detection and identification of neuropeptides. This study is premised on determining the most efficient neuropeptide extraction protocol and utilizing it to obtain endogenous neuropeptide profile of a principal central auditory nucleus and how it is impacted by deafening noise. In this work, four different neuropeptide extraction protocols from brain tissue samples were examined for efficiency in LC-MS/MS analysis. Overall, acidified methanol extraction was the most efficient, yielding over twenty neuropeptides with seventeen of them being novel. The work also revealed noise induced alterations in the neuropeptide levels and implicate them as potential therapeutic targets.
Abstract Format
html
Disciplines
Biochemistry, Biophysics, and Structural Biology
Language
English
Extent
76 pages
Rights Statement
Copyright is held by the author.
Digital Origin
Born digital
Recommended Citation
Al Mousa, Ahmad Hejji A, "Exploring Efficient Neuropeptide Extraction Methods for Quantitative Peptidomics and Application in a Principal Central Auditory Nucleus" (2025). Master's Theses. 349.
https://digscholarship.unco.edu/theses/349