Date of Award
2021
Document Type
Dissertation
Degree Name
Ph.D.
Organizational Unit
College of Natural Science and Mathematics, Biological Sciences
First Advisor
Erich J. Kushner
Second Advisor
Yan Qin
Third Advisor
Bruce Appel
Fourth Advisor
Michelle Knowles
Keywords
Collagen IV, Endothelial, Lysyl hydroxylase 3, Rab10, Rab25, Vasculature
Abstract
Collagen type IV (Col IV) is a basement membrane protein associated with early blood vessel morphogenesis and is essential for blood vessel stability. Defects in vascular Col IV deposition are the basis of heritable disorders, such as small vessel disease, marked by cerebral hemorrhage and drastically shorten lifespan. To date, little is known about how endothelial cells regulate the intracellular transport and selective secretion of Col IV in response to angiogenic cues, leaving a void in our understanding of this process. Our aim was to identify trafficking pathways that regulate Col IV deposition during angiogenic blood vessel development. We have identified the GTPase Rab10 as a major regulator of Col IV vesicular trafficking during vascular development using both in vitro imaging and biochemistry as well as in vivo models. Knockdown of Rab10 reduced de novo Col IV secretion in vitro and in vivo. Mechanistically, we determined that Rab10 is an indirect mediator of Col IV secretion partnering with atypical Rab25 to deliver the enzyme lysyl hydroxylase 3 (LH3) to Col IV-containing vesicles staged for secretion. Loss of Rab10 or Rab25 results in depletion of LH3 from Col IV-containing vesicles and rapid lysosomal degradation of Col IV. Furthermore, we demonstrate that Rab10 is Notch responsive, indicating a novel connection between permissive Notch-based blood vessel maturation programs and vesicle trafficking. Our results illustrate both a new trafficking-based component in the regulated secretion of Col IV and how this vesicle trafficking program interfaces with Notch signaling to fine-tune basement membrane secretion during blood vessel development.
Publication Statement
Copyright is held by the author. User is responsible for all copyright compliance.
Rights Holder
Stephen J.B. Gross
Provenance
Received from ProQuest
File Format
application/pdf
Language
en
File Size
116 pgs
Recommended Citation
Gross, Stephen J.B., "Notch Regulates Vascular Collagen IV Basement Membrane Through Modulation of Lysyl Hydroxylase 3 Trafficking" (2021). Electronic Theses and Dissertations. 1932.
https://digitalcommons.du.edu/etd/1932
Copyright date
2021
Discipline
Molecular biology