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Multiplexed DNA-Paint Imaging of the Heterogeneity of Late Endosome/Lysosome Protein Composition Charles Bond

Dissertations & Theses @ University of Pennsylvania Available online

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Format:
Book
Thesis/Dissertation
Author/Creator:
Bond, Charles, author.
Contributor:
University of Pennsylvania. Cell and Molecular Biology., degree granting institution.
Language:
English
Subjects (All):
Cellular biology.
Physiology.
Molecular biology.
Biochemistry.
0379.
0487.
0307.
0719.
Local Subjects:
Cellular biology.
Physiology.
Molecular biology.
Biochemistry.
0379.
0487.
0307.
0719.
Physical Description:
1 electronic resource (123 pages)
Contained In:
Dissertations Abstracts International 86-07B
Place of Publication:
Ann Arbor : ProQuest Dissertations and Theses, 2024
Language Note:
English
Summary:
Late endosomes/lysosomes (LELs) are crucial for numerous physiological processes and their dysfunction is linked to many diseases. Proteomic analyses have identified hundreds of LEL proteins, however, whether these proteins are uniformly present on each LEL, or if there are cell-type dependent LEL subpopulations with unique protein compositions is unclear. We employed quantitative, multiplexed DNA-PAINT super-resolution imaging to examine the distribution of seven key LEL proteins (LAMP1, LAMP2, CD63, Cathepsin D, TMEM192, NPC1 and LAMTOR4). While LAMP1, LAMP2 and Cathepsin D were abundant across LELs, marking a common population, most analyzed proteins were associated with specific LEL subpopulations. Our multiplexed imaging approach identified up to eight different LEL subpopulations based on their unique membrane protein composition. Additionally, our analysis of the spatial relationships between these subpopulations and mitochondria revealed a cell-type specific tendency for NPC1-positive LELs to be closely positioned to mitochondria. Our approach will be broadly applicable to determining organelle heterogeneity with single organelle resolution in many biological contexts.
Notes:
Source: Dissertations Abstracts International, Volume: 86-07, Section: B.
Advisors: Lakadamyali, Melike Committee members: Holzbaur, Erika; Chang, Yi-Wei; Mitchell, Claire; Ostap, E. Michael; Shalem, Ophir
Ph.D. University of Pennsylvania 2024
Local Notes:
School code: 0175
ISBN:
9798302185358
Access Restriction:
Restricted for use by site license

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