2026-07-23T17:11:31Zhttps://repisalud.isciii.es/rest/oai/requestoai:repisalud.isciii.es:20.500.12105/64842024-09-27T08:02:23Zcom_20.500.12105_19604com_20.500.12105_2051col_20.500.12105_19605
Repisalud
advisor
Isern, Joan
author
Gonzalez-Hernandez, Sara
2018-10-09T12:35:44Z
2018-10-09T12:35:44Z
2018-07-19
10.4321/repisalud.6484
http://hdl.handle.net/20.500.12105/6484
The study of the developmental origin of the coronary vasculature in mammals has been
subject of extensive research in recent years, and yet the signals regulating the formation of
the primary plexus, remain poorly understood. In this Doctoral Thesis the murine genetic tools
Nes-Gfp and Nes-CreERT2 lines have been used for studying coronary endothelium formation
during cardiac development, with special emphasis on the stages of remodeling and arteriovenous
maturation. The strong characterization of the Nes-Gfp allele during the intermediate
stages of cardiogenesis indicated that it is expressed in mesenchymal strains of the cardiac
bulb, pericytes and mural cells associated with the vessels (analogously to the pattern already
described in other organs), and unexpectedly and dynamic, its expression is also induced
robustly in the nascent coronary endothelium.
By taking advantage of this pattern of vascular expression and using whole-tissue
clarification imaging and confocal microscopy, we aim to characterize the process of sprouting
of the coronary vessels and arterial remodeling at unprecedented resolution. In addition,
thanks to the relative intensity of expression of the GFP reporter, in combination with the
Endomucin marker, it allowed us to discriminate between ventricular endocardial cells (where
GFP is barely expressed) and the coronary vascular plexus. By cytometry, we were able to
isolate both primary endothelial populations with great purity, from the same hearts and obtain
the corresponding transcriptional profiles, in different phases of their development: the initial
stage of angiogenic expansion (E13.5) and during the process of active remodeling of the
primary plexus (E17.5). Moreover, lineage tracing using the Nes-CreERT2 driver (which shares
the presence of regulatory sequences of the neural enhancer of the Nestin gene with the Nes-
Gfp line) allows us to selectively label the intramyocardial coronary plexus enriched in capillary
and arteriolar endothelium. Therefore, the combination of both genetic tools has allowed us to
isolate and define transcriptomically the three subpopulations of blood vascular cells present in
the heart (endocardium, intramyocardial arteries and subepicardial veins).
This strategy allowed us to identify new enriched factors in the arterial branch. Among
the differentially expressed genes, we have found several members of the SOX transcription
factors family, and especially Sox17. Interestingly, we detected nuclear expression of Sox17 in
the first endothelial cells of the plexus that migrated through the subepicardium. Later on, its
expression is progressively restricted in the domain of intramural Nes-GFP+ vessels, with an
arterial phenotype. Due to the robust co-localization observed between the Sox17+ population
and the GFP+ cells, we wonder if this transcription factor could be a direct transcriptional
activator of the neural enhancer present in the regulatory region of the Nes-Gfp allele, verifying
it with a series of assays in vitro. For all these reasons, we propose that the Nestin neural
enhancer could also act as a coronary arterial enhancer in certain phases of cardiac
development, and whose continued study could help to discover new molecular bases in this
important and relevant process for human health, given the high incidence of coronary disease
worldwide.
spa
enhancer
nestin
coronary vasculature
Sox17
Herramientas genéticas basadas en el enhancer neural de Nestina para la caracterización de las poblaciones endoteliales del corazón: papel del Sox17 en el desarrollo de la vasculatura coronaria
doctoral thesis
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Tesis Sara Gonzalez Hernandez.pdf
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Tesis Sara Gonzalez Hernandez.pdf.txt