Identification of differentially expressed genes after heat shock in isolated rat aorta.
1. In a previous study, we demonstrated that heat shock augments vascular contractility through the stress response. 2. The current study was designed to identify differentially expressed genes after heat shock by using a novel annealing control primer (ACP) system, which was developed recently to identify authentic genes. 3. Rat aortic rings were mounted in organ baths, exposed to 42 degrees C for 45 min and harvested 4 h after the end of heat shock. Total RNA were used for amplification by the reverse transcriptase-polymerase chain reaction (RT-PCR) with ACP system. Differentially amplified PCR products were sequenced, searched against the GenBank and confirmed by RT-PCR. 4. Genes for connective tissue growth factor, stress-inducible protein 1 and heat shock protein 25 were upregulated, whereas a gene for interferon regulatory factor 1 was downregulated. Immunohistochemistry revealed upregulation of the phosphorylated form of Hsp25 in aortic rings after heat shock. 5. These results suggest that phosphorylated Hsp25 plays a pivotal role in the augmentation of vascular contraction after heat shock.
Kim
Jee In
JI
Department of Pharmacology, Kyungpook National University School of Medicine, Daegu, Korea. inkim@knu.ac.kr
Kim
Su Jin
SJ
Jung
Sang Won
SW
Cho
Joon Yong
JY
Chung
Eun Jung
EJ
Kim
Moonkyu
M
Shin
Heung-Mook
HM
Kim
In Kyeom
IK
D000818:Animals; D001013:Aorta, Thoracic / Q000378:metabolism* / Q000503:physiopathology; D055513:Connective Tissue Growth Factor; D030541:Databases, Genetic; D015870:Gene Expression; D020869:Gene Expression Profiling / Q000379:methods; D055551:HSP27 Heat-Shock Proteins; D018882:Heat Stress Disorders / Q000235:genetics* / Q000378:metabolism / Q000503:physiopathology; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D006358:Hot Temperature; D017874:Immediate-Early Proteins / Q000235:genetics / Q000378:metabolism; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D050856:Interferon Regulatory Factor-1 / Q000235:genetics / Q000378:metabolism; D008297:Male; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014661:Vasoconstriction / Q000235:genetics*
C526316:CCN2 protein, rat; D055551:HSP27 Heat-Shock Proteins; D006360:Heat-Shock Proteins; C486114:Hspb1 protein, rat; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D050856:Interferon Regulatory Factor-1; C494259:Irf1 protein, rat; D009363:Neoplasm Proteins; D012333:RNA, Messenger; C489790:Stip1 protein, rat; D055513:Connective Tissue Growth Factor