Troponin I isoform expression in human and experimental atrial fibrillation.

BACKGROUND: Atrial fibrillation (AF) is accompanied by re-expression of fetal genes and activation of proteolytic enzymes. In this study both aspects were addressed with respect to troponin I (TnI) isoform expression. METHODS AND RESULTS: Western blotting and real-time reverse transcription-polymerase chain reaction were used to study TnI isoform expression in patients with paroxysmal or chronic AF and in goats after 1, 2, 4, 8, and 16 weeks of AF. In addition to cardiac TnI (cTnI), low expression of slow-twitch skeletal TnI (ssTnI) protein was found in 60% of patients in sinus rhythm or paroxysmal AF and in 8% of patients with chronic AF. In adult goat atrium, ssTnI protein expression was undetectable. Calcium-dependent degradation of cTnI protein was found in 1 or 2 of 6 animals after 1 to 4 weeks of AF. Although always low, ssTnI mRNA levels were significantly higher in patients who expressed ssTnI protein than in those who did not. Relative ssTnI mRNA expression was significantly lower in patients with paroxysmal AF and chronic AF than in those in sinus rhythm. In goats there was a tendency toward higher relative levels of ssTnI at the onset of AF followed by a normalization when AF had become sustained. CONCLUSIONS: Atrial re-expression of ssTnI during paroxysmal AF in patients and during the first 2 weeks of pacing-induced AF in goats does not seem to be part of the process of AF-associated cardiomyocyte dedifferentiation but seems to result from transient cardiomyocyte stress at the onset of AF.

Reverse structural and gap-junctional remodeling after prolonged atrial fibrillation in the goat.

BACKGROUND: Prolonged atrial fibrillation (AF) results in electrical, structural, and gap-junctional remodeling. We examined the reversibility of the changes in (ultra)structure and gap junctions. METHODS AND RESULTS: Four groups of goats were used: (1) sinus rhythm (SR), (2) 4 months' AF (4 mo AF), (3) 2 months' SR after 4 mo AF (2 mo post-AF), and (4) 4 months' SR after 4 mo AF (4 mo post-AF). Atria were characterized electrophysiologically, (ultra)structure was studied by light and electron microscopy, and structural and gap-junctional protein expression was studied by immunohistochemistry or Western blotting. The atrial effective refractory period had completely returned to normal values 2 mo post-AF. Induced AF episodes still lasted for minutes at 2 and 4 mo post-AF, compared with seconds in the SR group. Structural abnormalities were still present at 2 and 4 mo post-AF, although to a lesser extent. The increased atrial myocyte diameter was back to normal at 4 mo post-AF. The number of myocytes with severe myolysis had almost normalized 4 mo post-AF, whereas myocytes with mild myolysis remained significantly increased. Extracellular matrix area fraction after 4 mo AF was similar to SR. However, the extracellular matrix fraction per myocyte had increased after 4 mo AF and remained higher post-AF. Changes in expression of structural proteins were partially restored post-AF. The reduction of connexin 40 that was observed during AF was completely reversed at 4 mo post-AF. CONCLUSIONS: Recovery from structural remodeling after 4 mo AF is a slow process and is still incomplete 4 mo post-AF. Several months post-AF, the duration of AF episodes is still prolonged (minutes).

Cardiac gap junctions and connexins: their role in atrial fibrillation and potential as therapeutic targets.

In the heart, changes in velocity and in patterns of conduction of myocardial electrical activity can affect cardiac rhythm and the coordination of contraction. Abnormal electrical coupling between cardiomyocytes through gap junctions is, therefore, considered an important factor in various pathophysiologic conditions. In the present report we summarize the literature on gap junctions and their structural proteins, the connexins, in the normal and fibrillating atrium. Putative implications of the recently reported remodelling of atrial gap junctions for stability of the arrhythmia will be discussed. Also the reversibility of the remodelling process will be addressed in the light of a potentially new therapeutic target for controlling the progression of atrial fibrillation (AF).

Analysis of altered gene expression during sustained atrial fibrillation in the goat.

OBJECTIVE: Atrial fibrillation (AF) is characterised by electrical, gap junctional and structural remodelling. However, the underlying molecular mechanisms of these phenomena are largely unknown. To get more insight into atrial remodelling at the molecular level we have analysed changes in gene expression during sustained AF in the goat. METHODS: The differential display technique (DD) was used to identify genes differentially expressed during sustained AF (13.9 +/- 5.2 weeks) as compared to sinus rhythm (SR). Dot-blot analysis was performed to confirm the altered gene expression and to establish the changes in expression after 1, 2, 4, 8 and 16 weeks of AF. Immunohistochemistry and western blotting were used to validate the DD approach and to further characterise the changed expression of the beta-myosin heavy chain gene at the protein level. RESULTS: Of the approximately 125 fragments that showed changed expression levels during AF, 34 were cloned and sequenced. Twenty-one of these represented known genes involved in cardiomyocyte structure, metabolism, expression regulation, or differentiation status. The changed expression of 70% of the isolated clones could be confirmed by dot-blot analysis. In addition, time course analysis revealed different profiles of expression as well as transient re-expression of genes, e.g. the gene for hypoxia-inducible factor 1 alpha during the first week of AF. During sustained AF the frequency of cardiomyocytes expressing beta myosin heavy chain (beta MHC) increased from 21.8 +/- 2.1 to 47.9 +/- 2.5% (S.E.M.). The overall expression of MHC (alpha+beta) appeared to be down-regulated during AF. CONCLUSIONS: AF is accompanied by changes in expression of proteins involved in cellular structure, metabolism, gene expression regulation and (de-)differentiation. Most alterations in expression confirm or support the hypothesis of cardiomyocyte de-differentiation. Furthermore, the results suggest a role for ischemic stress in the early response of cardiomyocytes to AF, possibly via activation of hypoxia-inducible factor 1 alpha.