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Vascular smooth muscle investment of...

Subramanian, Ramiah.

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Vascular smooth muscle investment of zebrafish (Danio rerio) blood vessels.

Record Type:	Language materials, printed : Monograph/item
Title/Author:	Vascular smooth muscle investment of zebrafish (Danio rerio) blood vessels./
Author:	Subramanian, Ramiah.
Description:	79 p.
Notes:	Directors: Brant M. Weinstein; Tom Sargent.
Contained By:	Dissertation Abstracts International63-12B.
Subject:	Biology, Animal Physiology. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3075208
ISBN:	0493954295

Vascular smooth muscle investment of zebrafish (Danio rerio) blood vessels.
Subramanian, Ramiah.

Vascular smooth muscle investment of zebrafish (Danio rerio) blood vessels. - 79 p.

Directors: Brant M. Weinstein; Tom Sargent.

Thesis (Ph.D.)--The George Washington University, 2003.

The purpose of this research is to determine the source and time of origin of smooth muscle cells in the dorsal aorta in the zebrafish <italic>Danio rerio</italic> using molecular biological and morphological methods. I analyzed the temporal progression of vascular smooth muscle investment in the zebrafish by light and electron microscopy and by expression of molecular markers (Chapter 1). Results of these studies suggest that zebrafish form a smooth muscle invested dorsal aorta that is structurally similar to that of higher vertebrates, and that smooth muscle-associated molecular markers are expressed prior to obvious smooth muscle recruitment. I attempted to determine whether or not the endothelial cells that line blood vessels transdifferentiate into vascular smooth muscle cells during normal development (Chapter 2). To this end, I generated constructs in which the <italic>SM22</italic>α promoter drove expression of a red or green fluorescent marker gene. The injected constructs did express the fluorescent marker gene; however, the expression was not vascular specific. This would suggest that, at least in the zebrafish, additional regulatory elements not included in these constructs need to be present for vascular specific expression. Finally, in the hope of understanding the regulation of smooth muscle gene expression I cloned the transcription factor <italic> myocardin</italic> that is thought to regulate smooth muscle development and tried to ‘knock-down’ its expression (Chapter 3).1 obtained, using the polymerase chain reaction, a full-length cDNA clone for the zebrafish ortholog of <italic>myocardin</italic> and the sequence of this clone shows modest identity with mammalian homologues but the domains thought to be essential for the transcriptional activity are well conserved. Whole-mount in situ hybridization showed that the <italic>myocardin</italic> gene is expressed in the developing heart from about the 5-somite stage. I performed knock-down experiments utilizing antisense morpholino oligonucleotides directed against the zebrafish <italic> myocardin</italic> mRNA. Although myocardin is thought to be a regulator of the smooth muscle specific gene <italic>SM22</italic>α, injection of the morpholinos did not affect <italic>SM22</italic>α expression. This may be due to redundancy in the <italic>myocardin</italic> and related genes in zebrafish or lack of function of the injected morpholinos.

ISBN: 0493954295Subjects--Topical Terms:

1017835
Biology, Animal Physiology.

Vascular smooth muscle investment of zebrafish (Danio rerio) blood vessels.
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100 1 $a Subramanian, Ramiah. $3 1258433
245 1 0 $a Vascular smooth muscle investment of zebrafish (Danio rerio) blood vessels.
300 $a 79 p.
500 $a Directors: Brant M. Weinstein; Tom Sargent.
500 $a Source: Dissertation Abstracts International, Volume: 63-12, Section: B, page: 5657.
502 $a Thesis (Ph.D.)--The George Washington University, 2003.
520 $a The purpose of this research is to determine the source and time of origin of smooth muscle cells in the dorsal aorta in the zebrafish <italic>Danio rerio</italic> using molecular biological and morphological methods. I analyzed the temporal progression of vascular smooth muscle investment in the zebrafish by light and electron microscopy and by expression of molecular markers (Chapter 1). Results of these studies suggest that zebrafish form a smooth muscle invested dorsal aorta that is structurally similar to that of higher vertebrates, and that smooth muscle-associated molecular markers are expressed prior to obvious smooth muscle recruitment. I attempted to determine whether or not the endothelial cells that line blood vessels transdifferentiate into vascular smooth muscle cells during normal development (Chapter 2). To this end, I generated constructs in which the <italic>SM22</italic>α promoter drove expression of a red or green fluorescent marker gene. The injected constructs did express the fluorescent marker gene; however, the expression was not vascular specific. This would suggest that, at least in the zebrafish, additional regulatory elements not included in these constructs need to be present for vascular specific expression. Finally, in the hope of understanding the regulation of smooth muscle gene expression I cloned the transcription factor <italic> myocardin</italic> that is thought to regulate smooth muscle development and tried to ‘knock-down’ its expression (Chapter 3).1 obtained, using the polymerase chain reaction, a full-length cDNA clone for the zebrafish ortholog of <italic>myocardin</italic> and the sequence of this clone shows modest identity with mammalian homologues but the domains thought to be essential for the transcriptional activity are well conserved. Whole-mount in situ hybridization showed that the <italic>myocardin</italic> gene is expressed in the developing heart from about the 5-somite stage. I performed knock-down experiments utilizing antisense morpholino oligonucleotides directed against the zebrafish <italic> myocardin</italic> mRNA. Although myocardin is thought to be a regulator of the smooth muscle specific gene <italic>SM22</italic>α, injection of the morpholinos did not affect <italic>SM22</italic>α expression. This may be due to redundancy in the <italic>myocardin</italic> and related genes in zebrafish or lack of function of the injected morpholinos.
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