Selection of genes associated with variations in the Circle of Willis in gerbils using suppression subtractive hybridization

Deformities in the Circle of Willis (CoW) can significantly increase the risk of cerebrovascular disease in humans. However, the molecular mechanisms underlying these deformities have not been understood. Based on our previous studies, variations in the CoW of gerbils are hereditary. A normal CoW is...

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Published in:PloS one 2015-05, Vol.10 (5), p.e0127355-e0127355
Main Authors: Li, Zhenkun, Huo, Xueyun, Zhang, Shuangyue, Lu, Jing, Li, Changlong, Guo, Meng, Fu, Rui, He, Zhengming, Du, Xiaoyan, Chen, Zhenwen
Format: Article
Language:English
Subjects:
Analysis
Angiogenesis
Animal sciences
Animals
Blood vessels
Cardiovascular disease
Carotid arteries
Cattle
Cerebrovascular disorders
Circle of Willis - abnormalities
Cloning
Cystatin C - genetics
Deformation mechanisms
Expressed sequence tags
Gene sequencing
Genes
Genetic aspects
Genetic Association Studies
Gerbillinae
Glutathione Peroxidase - genetics
GTP-Binding Protein alpha Subunits, Gs - genetics
Health risks
Hybridization
Ischemia
Laboratory animals
Molecular modelling
Molecular Sequence Data
Mutation
Polymerase chain reaction
Profilins - genetics
Risk factors
Southern blotting
Subtractive Hybridization Techniques
Variation
Vascular endothelial growth factor
Veins & arteries
Western blotting
Zoology
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cited_by cdi_FETCH-LOGICAL-c692t-f441eb32a26f743c16c17c2777da54c4f3c462d69924737d6f1b600bc081bcaf3
cites cdi_FETCH-LOGICAL-c692t-f441eb32a26f743c16c17c2777da54c4f3c462d69924737d6f1b600bc081bcaf3
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container_issue 5
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container_title PloS one
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creator Li, Zhenkun
Huo, Xueyun
Zhang, Shuangyue
Lu, Jing
Li, Changlong
Guo, Meng
Fu, Rui
He, Zhengming
Du, Xiaoyan
Chen, Zhenwen
description Deformities in the Circle of Willis (CoW) can significantly increase the risk of cerebrovascular disease in humans. However, the molecular mechanisms underlying these deformities have not been understood. Based on our previous studies, variations in the CoW of gerbils are hereditary. A normal CoW is observed in approximately 60% of gerbils, a percentage that also applies to humans. Thus, gerbil is an ideal experimental model for studying variations in the CoW. To study the mechanisms underlying these variations, we selected genes associated with different types of the CoW using suppression subtractive hybridization (SSH). After evaluating the efficiency of SSH using quantitative real-time polymerase chain reaction (qPCR) on subtracted and unsubtracted cDNA and Southern blotting on SSH PCR products, 12 SSH libraries were established. We identified 4 genes (CST3, GNAS, GPx4 and PFN2) associated with variations in the CoW. These genes were identified with qPCR and Western blotting using 70 expressed sequence tags from the SSH libraries. Cloning and sequencing allowed us to demonstrate that the 4 genes were closely related to mouse genes. We may assume that these 4 genes play an important role in the development of variations in the CoW. This study provides a foundation for further research of genes related to development of variations in the CoW and the mechanisms of dysmorphosis of cerebral vessels.
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However, the molecular mechanisms underlying these deformities have not been understood. Based on our previous studies, variations in the CoW of gerbils are hereditary. A normal CoW is observed in approximately 60% of gerbils, a percentage that also applies to humans. Thus, gerbil is an ideal experimental model for studying variations in the CoW. To study the mechanisms underlying these variations, we selected genes associated with different types of the CoW using suppression subtractive hybridization (SSH). After evaluating the efficiency of SSH using quantitative real-time polymerase chain reaction (qPCR) on subtracted and unsubtracted cDNA and Southern blotting on SSH PCR products, 12 SSH libraries were established. We identified 4 genes (CST3, GNAS, GPx4 and PFN2) associated with variations in the CoW. These genes were identified with qPCR and Western blotting using 70 expressed sequence tags from the SSH libraries. Cloning and sequencing allowed us to demonstrate that the 4 genes were closely related to mouse genes. We may assume that these 4 genes play an important role in the development of variations in the CoW. 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However, the molecular mechanisms underlying these deformities have not been understood. Based on our previous studies, variations in the CoW of gerbils are hereditary. A normal CoW is observed in approximately 60% of gerbils, a percentage that also applies to humans. Thus, gerbil is an ideal experimental model for studying variations in the CoW. To study the mechanisms underlying these variations, we selected genes associated with different types of the CoW using suppression subtractive hybridization (SSH). After evaluating the efficiency of SSH using quantitative real-time polymerase chain reaction (qPCR) on subtracted and unsubtracted cDNA and Southern blotting on SSH PCR products, 12 SSH libraries were established. We identified 4 genes (CST3, GNAS, GPx4 and PFN2) associated with variations in the CoW. These genes were identified with qPCR and Western blotting using 70 expressed sequence tags from the SSH libraries. Cloning and sequencing allowed us to demonstrate that the 4 genes were closely related to mouse genes. We may assume that these 4 genes play an important role in the development of variations in the CoW. This study provides a foundation for further research of genes related to development of variations in the CoW and the mechanisms of dysmorphosis of cerebral vessels.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>25973917</pmid><doi>10.1371/journal.pone.0127355</doi><oa>free_for_read</oa></addata></record>
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1932-6203
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subjects Analysis
Angiogenesis
Animal sciences
Animals
Blood vessels
Cardiovascular disease
Carotid arteries
Cattle
Cerebrovascular disorders
Circle of Willis - abnormalities
Cloning
Cystatin C - genetics
Deformation mechanisms
Expressed sequence tags
Gene sequencing
Genes
Genetic aspects
Genetic Association Studies
Gerbillinae
Glutathione Peroxidase - genetics
GTP-Binding Protein alpha Subunits, Gs - genetics
Health risks
Hybridization
Ischemia
Laboratory animals
Molecular modelling
Molecular Sequence Data
Mutation
Polymerase chain reaction
Profilins - genetics
Risk factors
Southern blotting
Subtractive Hybridization Techniques
Variation
Vascular endothelial growth factor
Veins & arteries
Western blotting
Zoology
title Selection of genes associated with variations in the Circle of Willis in gerbils using suppression subtractive hybridization
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