Matching Items (4)
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- All Subjects: proteomics
- Language: English
- Creators: Borges, Chad
- Creators: College of Integrative Sciences and Arts
Description
White-nose syndrome (WNS) is a fungal disease that infects hibernating bats of multiple species across large portions of eastern North America. To date, WNS has been responsible for the deaths of over seven million bats. It is not yet known why certain species are able to resist infection. Since the fungus invades the skin and some resistant species show no signs of the characteristic cutaneous lesions, it seems likely that resistant species contain specific defense mechanisms within their skin, such as antimicrobial peptides (AMPs) and other immunologically relevant proteins expressed by specific cell types or as secreted soluble components. Proteomics could be a useful tool for understanding differences in susceptibility, and could help identify AMPs that could be synthesized and used as control agents against the spread of the causative fungus. This study is the first to optimize proteomics methods for bat wing tissues in order to compare the skin proteomes of species variably impacted by WNS, including those of two endangered species. Further tests are planned to investigate methods of increasing protein yield without altering the size of the tissue sample collected, as well as the analysis of mass spectrometry data from processed skin tissues of five bat species differentially affected by WNS.
ContributorsPatrose, Reena Paulene (Author) / Moore, Marianne (Thesis director) / Steele, Kelly (Committee member) / College of Integrative Sciences and Arts (Contributor) / Barrett, The Honors College (Contributor)
Created2018-05
Description
White-nose syndrome (WNS) is a cutaneous fungal infection caused by Pseudogymnoascus destructans (Pd) which was first observed in the United States in 2006. Pd infects bats during hibernation and leads to the development of cutaneous lesions and behavioral changes that can result in the animal's death. This study generated the first complete bat skin proteome for the WNS resistant gray bat (Myotis grisescens) to optimize sample preparation methods and identify immune proteins that may signal resistance. Wing tissue was collected from a female gray bat and processed in a Barocycler using 4M or 8M urea followed by an in-gel trypsin digestion of pooled samples and processing of separate samples without digestion specifically to capture and identify small antimicrobial peptides. Both undigested and digested samples were analyzed using a Thermo Fisher LTQ Orbitrap Velos mass spectrometer and interpreted using PEAKS software. A total of 29 immune proteins were identified including the antimicrobial peptide dermcidin. This method will be applied to a larger range of samples from five species variably impacted by WNS to compare skin proteomes with the aim of identifying immune proteins that are responsible for resistance at the barrier where Pd invades.
ContributorsBoone, Brianna Marie (Author) / Moore, Marianne (Thesis director) / Steele, Kelly (Committee member) / College of Integrative Sciences and Arts (Contributor) / Department of Psychology (Contributor) / Barrett, The Honors College (Contributor)
Created2017-05
Description
Alzheimer’s Disease (AD) is the most common form of dementia affecting the population over the age of 65. AD is characterized clinically by increasing difficulty with memory and language, resulting in a loss of independence. This is due to the presence of two characteristic protein aggregates in the brain: extracellular amyloid plaques and intracellular neurofibrillary tangles (NFTs). Utilizing multiplexed immunofluorescence and dimensional reduction analysis the types of cells present in the hippocampus, the region of the brain most affected by AD, can be explored. Understanding the kinds of cell subtypes present, the mechanism behind how AD develops can be explored. Multiplexed IF was performed on human hippocampus FFPE tissues to detect a total of 37 proteins. Dimensional reduction analysis was performed to identify the four major cell types in the brain: neurons, oligodendrocytes, astrocytes, and microglia. After identifying each cell type, further dimensional reduction analysis was performed within each cell type to identify cell subtypes. A total of 21 neuron, 41 oligodendrocyte, 20 astrocyte, and 22 microglia subtypes were identified. The location of cell subtypes in each region of the hippocampal formation was found to match previous reports, further validating the findings of this project.
ContributorsEllison, Mischa A (Author) / Guo, Jia (Thesis advisor) / Borges, Chad (Committee member) / Mastroeni, Diego (Committee member) / Arizona State University (Publisher)
Created2024
Description
Measurements of different molecular species from single cells have the potential to reveal cell-to-cell variations, which are precluded by population-based measurements. An increasing percentage of researches have been focused on proteins, for its central roles in biological processes. Immunofluorescence (IF) has been a well-established protein analysis platform. To gain comprehensive insights into cell biology and diagnostic pathology, a crucial direction would be to increase the multiplexity of current single cell protein analysis technologies.
An azide-based chemical cleavable linker has been introduced to design and synthesis novel fluorescent probes. These probes allow cyclic immunofluorescence staining which leads to the feasibility of highly multiplexed single cell in situ protein profiling. These highly multiplexed imaging-based platforms have the potential to quantify more than 100 protein targets in cultured cells and more than 50 protein targets in single cells in tissues.
This approach has been successfully applied in formalin-fixed paraffin-embedded (FFPE) brain tissues. Multiplexed protein expression level results reveal neuronal heterogeneity in the human hippocampus.
An azide-based chemical cleavable linker has been introduced to design and synthesis novel fluorescent probes. These probes allow cyclic immunofluorescence staining which leads to the feasibility of highly multiplexed single cell in situ protein profiling. These highly multiplexed imaging-based platforms have the potential to quantify more than 100 protein targets in cultured cells and more than 50 protein targets in single cells in tissues.
This approach has been successfully applied in formalin-fixed paraffin-embedded (FFPE) brain tissues. Multiplexed protein expression level results reveal neuronal heterogeneity in the human hippocampus.
ContributorsLiao, Renjie (Author) / Guo, Jia (Thesis advisor) / Borges, Chad (Committee member) / Liu, Yan (Committee member) / Arizona State University (Publisher)
Created2019