Measurement of cardiac gene expression by reverse transcription polymerase chain reaction (RT-PCR). 2007

Nicola King
Bristol Heart Institute, Department Clinical Science Medicine at South Bristol, Faculty of Medicine and Dentistry, University of Bristol, UK.

Two methods applicable to the measurement of cardiac gene expression by reverse transcription polymerase chain reaction (RT-PCR) are presented. Each method describes a specific technique and includes information on how to optimize the experiments and how to trouble-shoot any problems. The first method illustrates how conventional RT-PCR was used to detect expression of a specific gene in rat heart. The second method explains how to investigate the amount of gene expressed under different conditions utilizing the new technique of quantitative real-time RT-PCR. More specifically, this second method was used to investigate the effect of a relevant cardiac insult, ischemia reperfusion, on gene expression, although the technique could be readily adapted to other cardiac insults or to compare different hearts or treatments. The gene investigated in these examples was SNAT3, which is responsible for the cotransport of sodium predominantly with glutamine in exchange for hydrogen ions. The results suggested that a 20-fold increase in SNAT3 expression occurs during ischemia, which reduced to baseline levels on reperfusion.

UI MeSH Term Description Entries
D009206 Myocardium The muscle tissue of the HEART. It is composed of striated, involuntary muscle cells (MYOCYTES, CARDIAC) connected to form the contractile pump to generate blood flow. Muscle, Cardiac,Muscle, Heart,Cardiac Muscle,Myocardia,Cardiac Muscles,Heart Muscle,Heart Muscles,Muscles, Cardiac,Muscles, Heart
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D012333 RNA, Messenger RNA sequences that serve as templates for protein synthesis. Bacterial mRNAs are generally primary transcripts in that they do not require post-transcriptional processing. Eukaryotic mRNA is synthesized in the nucleus and must be exported to the cytoplasm for translation. Most eukaryotic mRNAs have a sequence of polyadenylic acid at the 3' end, referred to as the poly(A) tail. The function of this tail is not known for certain, but it may play a role in the export of mature mRNA from the nucleus as well as in helping stabilize some mRNA molecules by retarding their degradation in the cytoplasm. Messenger RNA,Messenger RNA, Polyadenylated,Poly(A) Tail,Poly(A)+ RNA,Poly(A)+ mRNA,RNA, Messenger, Polyadenylated,RNA, Polyadenylated,mRNA,mRNA, Non-Polyadenylated,mRNA, Polyadenylated,Non-Polyadenylated mRNA,Poly(A) RNA,Polyadenylated mRNA,Non Polyadenylated mRNA,Polyadenylated Messenger RNA,Polyadenylated RNA,RNA, Polyadenylated Messenger,mRNA, Non Polyadenylated
D015427 Reperfusion Injury Adverse functional, metabolic, or structural changes in tissues that result from the restoration of blood flow to the tissue (REPERFUSION) following ISCHEMIA. Ischemia-Reperfusion Injury,Injury, Ischemia-Reperfusion,Injury, Reperfusion,Reperfusion Damage,Damage, Reperfusion,Injury, Ischemia Reperfusion,Ischemia Reperfusion Injury,Ischemia-Reperfusion Injuries,Reperfusion Damages,Reperfusion Injuries
D015870 Gene Expression The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION. Expression, Gene,Expressions, Gene,Gene Expressions
D051381 Rats The common name for the genus Rattus. Rattus,Rats, Laboratory,Rats, Norway,Rattus norvegicus,Laboratory Rat,Laboratory Rats,Norway Rat,Norway Rats,Rat,Rat, Laboratory,Rat, Norway,norvegicus, Rattus
D020133 Reverse Transcriptase Polymerase Chain Reaction A variation of the PCR technique in which cDNA is made from RNA via reverse transcription. The resultant cDNA is then amplified using standard PCR protocols. Polymerase Chain Reaction, Reverse Transcriptase,Reverse Transcriptase PCR,PCR, Reverse Transcriptase,Transcriptase PCR, Reverse
D026921 Amino Acid Transport Systems, Neutral Amino acid transporter systems capable of transporting neutral amino acids (AMINO ACIDS, NEUTRAL). Neutral Amino Acid Transport Systems,Neutral Amino Acid Transporters,Zwitterionic Amino Acid Transport Systems,Na+-Independent Neutral Amino Acid Transporter,Neutral Amino Acid Transport Proteins,Sodium Dependent Neutral Amino Acid Transport Proteins,Sodium Dependent Neutral Amino Acid Transporters,Sodium Independent Neutral Amino Acid Transport Proteins,Sodium Independent Neutral Amino Acid Transporters,Zwitterionic Amino Acid Transport Proteins,Zwitterionic Amino Acid Transporters,Na+ Independent Neutral Amino Acid Transporter

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