Reprinted with permission from 801

JOURNAL OF CARDIOVASCULAR ELECTROPHYSIOLOGY, Volume 13, No. 8, August 2002

Copyright ©2002 by Futura Publishing Company, Inc., Armonk, NY 10504-0418

Aging-Related Increase to Inducible Atrial Fibrillation

in the Rat Model
HIDEKI HAYASHI, M.D., CHARLES WANG, M.D., YASUSHI MIYAUCHI, M.D.,
CHIKAYA OMICHI, M.D., HUI-NAM PAK, M.D., SHENGMEI ZHOU, M.D.,
TOSHIHIKO OHARA, M.D., WILLIAM J. MANDEL, M.D., SHIEN-FONG LIN, PH.D.,
MICHAEL C. FISHBEIN, M.D., PENG-SHENG CHEN, M.D.,
and HRAYR S. KARAGUEUZIAN, PH.D.
From the Division of Cardiology, Department of Medicine, Cedars-Sinai Medical Center, and the Department of Pathology and
Laboratory Medicine, UCLA School of Medicine, Los Angeles, California

Aging and Atrial Fibrillation. Introduction: Aging is associated with atrial interstitial Ž brosis and
increased incidence of atrial Ž brillation (AF). We hypothesized that aged rats are suitable for study of
aging-related AF and that partial atrial cellular uncoupling induced with heptanol in young rats mimics
aging-related AF.
Methods and Results: Interatrial conduction time and atrial response to burst atrial pacing were
evaluated in 11 young (2–3 months) and 12 old (22–24 months) male rats (Fisher 344) in the Langendorff-
perfused setting. At baseline, sustained ( > 30 sec) atrial tachycardia (AT) and AF were induced in 10 of
12 and in 7 of 12 old rats, respectively. No such arrhythmias could be induced in the young rats. Old rats
had signiŽ cantly (P < 0.01) longer interatrial conduction time and P wave durations than the young rats.
Burst pacing failed to induce AT and AF in all 11 young rats studied. The effects of heptanol 2 to 10 mM
were studied in both groups. Heptanol 2 to 5 mM promoted inducible AT in all 5 young rats studied;
however, when its concentration was raised to 10 mM, AT could no longer be induced in any of the 5 young
rats. No AF could be induced in any of the 5 young rats at heptanol concentrations of 2 to 10 mM. In the
old rats, AF could still be induced during perfusion of 2 mM heptanol. However, when its concentration
was raised to 5 and 10 mM, AF could not be induced in any of the 6 old rats studied. Optical mapping using
a potentiometric dye showed a periodic single wavefront of activation during AT in both groups and 2 to
4 independent wavefronts propagating in different directions during AF in the old rats. Histology revealed
a signiŽ cant increase in interstitial atrial Ž brosis (P < 0.01), atrial cell size (P < 0.05), and heart weight
in old versus young rats. Fibrosis in the old rats was highly heterogeneous.
Conclusion: The rat model is suitable for study of aging-related AF. Uniform partial atrial cellular
uncoupling with heptanol perfusion in the young rats, although promoting inducible AT, does not mimic
aging-related AF. The results suggest that heterogeneous atrial interstitial Ž brosis and atrial cell hyper-
trophy might contribute to the aging-related increase in atrial conduction slowing, conduction block, and
inducible AF in the old rat model. (J Cardiovasc Electrophysiol, Vol. 13, pp. 801-808, August 2002)

aging, atrial Ž brillation, cellular uncoupling, optical mapping, Ž brosis, heptanol, reentry

Introduction duration once it is induced.6 Atrial conduction slowing was

Atrial Ž brillation (AF) is the most common cardiac ar-
rhythmia seen in clinical practice.1 - 3 The prevalence of AF
increases with aging (i.e., about 0.1% at age 40 years, 6% at
65 years, and 10% at 80 years or older).4 Increased nonuni-
form atrial interstitial Ž brosis that characteristically devel-
ops during aging5 could promote conduction slowing and
wavefront breakups and may maintain AF for a longer
This study was supported by fellowship grants from the Cedars-Sinai
ECHO Foundation Award, Pauline and Harold Price Endowment, Piansky
Endowment, NIH SCOR Grant in Sudden Death (P50-HL-52319), AHA
Western States AfŽ liate Grant-in-Aid (0255937Y), UC-TRDRP
9RT-0041, and the Ralph M. Parsons Foundation, Los Angeles, California.
Address for correspondence: Hrayr S. Karagueuzian, Ph.D., Division of
Cardiology, Cedars Sinai Medical Center, Davis Research Building, Room
6066, 8700 Beverly Boulevard, Los Angeles, CA 90048. Fax: 310-423-
0291; E-mail: karagueuzian@cshs.org
documented clinically in patients with paroxysmal AF.7
However, at present there is no animal model that would
allow for a systematic search for a factor(s) that might
contribute to the aging-related increase in AF incidence.
Taking advantage of the systematic availability of aged
rodents from the National Institute on Aging, the Ž rst aim of
the present study was to determine if the rat model is
suitable for studying aging-related increase in inducible AF.
The second aim of the study was to test the hypothesis that
the partial cellular uncoupling of atrial cells in young rats
exposed to heptanol8 mimics the aging-related increase in
AF vulnerability. The results show that the rat model is
suitable for study of the aging-related increase in AF vul-
nerability and that partial cellular uncoupling of atria in
young rats with heptanol perfusion does not mimic the
aging-related increase in AF vulnerability.
Methods

Manuscript received 26 April 2002; Accepted for publication 12 June The research protocol was approved by the Institutional
2002. Animal Care and Use Committee of Cedars-Sinai Medical
802 Journal of Cardiovascular Electrophysiology Vol. 13, No. 8, August 2002
Center and followed the guidelines of American Heart As-
sociation. Twenty-three male Fischer 344 rats, 11 young
(age 2–3 months) and 12 old (age 22–24 months) were
anesthetized with an intraperitoneal injection of ketamine
100 mg/kg and xylazine 10 mg/kg.
In Situ Studies
The body surface ECG using six-limb leads was re-
corded in the closed chest anesthetized state for 5 minutes.
The duration of the P wave and the corrected duration of the
P wave (i.e., the duration of the P wave divided by square
root of the previous PP interval) were measured in both
groups on lead I.
In Vitro Studies
Langendorff-perfused rat hearts
The hearts were isolated and perfused (5 mL/min)
through the aorta with oxygenated Tyrode’s solution at
36.5° 6 1°C. The composition of the Tyrode’s solution was
as follows (in mM): NaCl 125, KCl 4.5, MgCl2 0.5, CaCl2
0.54, NaH2PO4 1.2, NaHCO3 24, glucose 5.5, and albumin
50 mg/L. The entire isolated hearts also were superfused
with warm oxygenated Tyrode’s solution at a rate of 20
mL/min. Two Te on-coated (except at their tips) silver
(0.35-mm diameter) bipolar electrodes (interelectrode dis-
tance 1 mm) were sutured on the right atrium (RA), two on
the left atrium (LA), and one on the left ventricle. The
distance (center to center) between the two RA bipolar
electrograms was 1 mm.
Interatrial conduction time and effective refractory period
The interatrial conduction time (IACT) was measured
during RA pacing at cycle lengths (CLs) of 300, 200, 100,
and 50 msec. The conduction time between two electrodes
10 mm apart, one placed on the RA appendage and the
second on the LA appendage, was taken as the IACT. The
effective refractory period (ERP) of the RA was measured
by the S2 extrastimulus method using eight regularly paced
beats at CL 5 200 msec using twice threshold current at
2-msec duration.
Atrial vulnerability
Burst RA pacing (CLs of 100 to 10 msec, in 10-msec
decrements) for 3-second duration with twice diastolic
threshold current was used to test for AF vulnerability.
Testing for atrial tachycardia (AT)/AF vulnerability with
rapid RA pacing was attempted Ž ve times at each pacing CL
in each rat in both groups. Up to 50 atrial bursts were
attempted to induce AF. The induced rhythm was deŽ ned as
sustained AT when the atrial bipolar electrogram morphol-
ogy was uniform, the activation interval periodic with an
isoelectric interval, and the rhythm lasted for .30 seconds.
AF was said to be present when the atrial bipolar electro-
gram morphology was nonuniform and the activation inter-
vals were irregular, showed no isoelectric interval, and had
a cycle length #50 msec. When an AT was Ž rst induced,
the rapid pacing protocol for AF induction continued after
the AT terminated spontaneously (up to 50 burst pacing
trials in each rat).
Effects of heptanol
The effects of increasing concentrations (2 to 10 mM) of
heptanol (Sigma) on IACT, ERP, and AF vulnerability were
tested after baseline measurements were made in 5 young
and 6 old rats. Heptanol was simultaneously perfused
through the aorta at a rate of 5 mL/min and superfused in the
tissue chamber at a rate of 20 mL/min.
Optical mapping
The hearts were stained with 0.3 mM of the voltage-
sensitive dye di-4-ANEPPS (Molecular Probes Inc., Eu-
gene, OR, USA), which was injected through the aorta. The
hearts were illuminated with a solid-state, frequency-dou-
bled laser (Verdi; Coherent Inc., Santa Clara, CA, USA) at
a wavelength of 532 nm. The emitted  uorescence was
acquired with a charge coupled device (CCD) camera (CA-
D1-0128T; Dalsa Inc., Waterloo, Ontario, Canada) through
a 600-nm long-pass Ž lter. The camera acquired the data
from 128 3 128 sites simultaneously over a 20 3 20 mm2
area with a time resolution of 2.4 msec per frame. In order
to allow visualization of activation patterns in greater detail,
only a portion of the mapped region was selected for pre-
sentation in the Ž gures. The image frames were color coded
and animated according to the amplitudes of the optical
membrane potential. The digital images were transferred to
a personal computer with a frame grabber (Roadrunner,
Bit ow Inc.). No electromechanical uncoupling agent was
used in the study.9 , 1 0
Anatomic-histologic analyses
The hearts of the young and the old rats were weighed
after conclusion of the in vitro studies. The hearts then were
Ž xed in 4% buffered formalin for 1 hour and then placed in
70% alcohol. Transmural sections of 5 mm were cut per-
pendicularly to the epicardium from parafŽ n-embedded tis-
sue blocks and mounted onto slides. The sections were
stained with either trichrome or hematoxylin and eosin
stains.1 1 The slides were examined by light microscopy at
4003 magniŽ cation. The ratio of the area occupied by
interstitial Ž brosis to the total area was determined in each
trichrome-stained sample using a computer-assisted image
analysis software (Image-Pro Plus 4.0). Results were given
as percentage Ž brosis of total atrial surface area. Five sec-
tions were made in each RA and LA in both groups. The
diameter of the atrial myocytes at the level of the nucleus in
longitudinal sections also was measured in the hematoxylin
and eosin stained sections. Twenty cells in each of the Ž ve
RA and LA samples from each rat of both groups were
sampled. The mean of the standard deviation (SD) of the
size of the myocytes and percentage Ž brosis at each of the
four atrial sites were calculated and used as indices of
heterogeneity of atrial myocyte size and of atrial Ž brous
tissue distribution, respectively. Total heart weights were
measured.
Statistical Analysis
All statistical analyses were performed using GB-Stat.
Data are expressed as mean 6 SD. When appropriate,
statistical analyses were performed using Student’s t-test,
Chi-square test, and two-way repeated measures of analysis
 Hayashi et al. Aging and Atrial Fibrillation 803

Figure 1. Body surface lead I ECG in an anesthetized

(A) young rat and (B) old rat in sinus rhythm. P wave
(two upward arrows) duration in the old rat is longer
than in the young rat (see superimposed P waves in
panel C). (D) P-P intervals, (E) P wave duration
during normal sinus rhythm, and (F) corrected P
wave duration (P wave duration divided by root
square of previous P-P interval) in young and old
rats. *P , 0.01.

of variance. P , 0.05 was considered statistically signiŽ - 2B and 2D). However, the longest IACT in the young rats
cant. induced by heptanol at pacing CL 5 50 msec did not exceed
46 msec. Testing of heptanol 50 mM in two young and two
Results
old rats resulted in complete loss of capture and failure of
In Vivo Studies atrial activation. Therefore, we tested AT/AF vulnerability
in the presence of heptanol 2 to 10 mM.
The body and heart weights of the old rats were signif-
icantly (P , 0.01) heavier than those of the young rats:
420 6 46 g versus 292 6 46 g and 1.8 6 0.3 g versus 1.4 6
0.3 g, respectively. Mean sinus CL was not signiŽ cantly
different between the old (250 6 49 msec) and the young
(225 6 34 msec) rats. However, mean P wave duration and
corrected P wave duration were signiŽ cantly (P , 0.05)
longer in the old compared with the young rats (32.5 6 3.8
msec vs 24.6 6 1.2 msec, respectively) (Fig. 1).
In Vitro Studies
ERP and excitability
Diastolic excitability threshold and RA ERP were not
signiŽ cantly different between the old and the young rats
(0.27 6 0.17 mA vs 0.23 6 0.18 mA and 19.3 6 7.6 msec
vs 19.0 6 4.8 msec, respectively). Heptanol 2 mM had no
effect on either ERP or excitability. However, at higher (.2
mM) concentrations, it caused loss of regular captures lead-
ing to complete failure of impulse propagation.
Interatrial conduction time
IACT in the young rats (Fig. 2A) was signiŽ cantly faster
than in the old rats (Fig. 2C) (23.8 6 4.6 msec vs 54.4 6
19.9 msec at pacing CL 5 300 msec and 24.3 6 4.9 msec
vs 57.6 6 22.9 msec at pacing CL 5 200 msec, P , 0.05
for both comparisons). IACT conduction slowing was rate
dependent (Figs. 2A and 2C). In the old rats, 2:1 conduction
block and complete failure of RA to LA propagation devel-
oped at CL 5 100 and 50 msec, respectively (Fig. 2C). No Figure 2. Effects of progressively shorter right atrial (RA) pacing cycle
lengths (PCL) on interatrial conduction time (IACT). Arrows point to the
such conduction alterations developed in the young rats
direction of propagation from the RA to the left atrium (LA). (A) IACT in
(Fig. 2A). However, signiŽ cant (P , 0.05) rate-dependent
a young rat at baseline and (B) during 2 mM heptanol perfusion. (C) IACT
IACT slowing developed in the young rats (23.8 6 4.6 msec in an old rat at baseline and (D) during 2 mM heptanol. Note 2:1
at pacing CL 5 300 msec vs 30.9 6 5.9 msec at CL 5 50 conduction block at PCL 5 50 msec in the old rat at baseline (C) and
msec) (Fig. 2A). Heptanol 2 mM ampliŽ ed rate-dependent complete IACT block at PCL 5 100 and 50 msec after heptanol (D). Top
IACT slowing in both groups, causing loss of regular cap- tracing in each panel is pacing stimulus artifact. Numbers denote the
ture in the old rats at pacing CL 5 100 and 50 msec (Figs. PCLs. Beg 5 bipolar electrogram.
804 Journal of Cardiovascular Electrophysiology Vol. 13, No. 8, August 2002

heptanol 2 mM was present, burst atrial pacing reduced AT

induction from 6 of 6 to 4 of 6 rats. Further increases of
heptanol concentrations to 5 and 10 mM prevented AT
induction in all six old rats (Figs. 4E and 4F). The effects of
increasing heptanol concentrations on inducible AF were
studied in six old rats. Whereas heptanol 2 mM increased
AF induction from 3 of 6 to 4 of 6, increasing its concen-
tration to 5 and 10 mM prevented AF induction in all six old
rats (biphasic effect). AF CL at baseline was 38.3 6 1.5
msec; after 2 mM heptanol, CL was 32.3 6 6.8 msec (P 5
NS).
Optical mapping
During AT, a single large activation wavefront periodi-
cally invaded the Ž eld of optical view in both young and old
rats (Fig. 5A). The propagation was sequential, proceeding
from one side of the optical Ž eld of view to the other (Fig.
5A). Similarly, AT induced in the presence of heptanol in
Figure 3. Effects of burst atrial pacing on atrial rhythm in a young rat. (A)
Burst atrial pacing at baseline fails to induce arrhythmia. Normal sinus
rhythm resumes after the burst. (B) Burst atrial pacing in an old rat
induces atrial tachycardia (AT) at a cycle length of 79 ms. (C) Induction
of atrial Ž brillation (AF) in another old rat. LA Beg 5 LA bipolar
electrogram; LV Beg 5 left ventricular bipolar electrogram; RA Beg 5
right atrial bipolar electrogram; Stim 5 stimulation artifact.

Testing of AF vulnerabilities
All isolated-perfused rat hearts in both groups were in
normal sinus rhythm, showing sequential activation of the
RA Ž rst, then the LA, and then the ventricles (Fig. 3A).
Burst atrial pacing failed to induce atrial tachyarrhythmia in
all 11 young rats (Fig. 3A). Figure 3A shows the resumption
of normal sinus rhythm at the end of burst atrial pacing in a
young rat. In contrast, burst atrial pacing induced AT in 10
of 12 old rats and AF in 7 of 12 old rats (Figs. 3B and 3C).
Figure 3B shows burst atrial pacing-inducing AT in one old
rat with complete AV block, and Figure 3C shows AF
induced in an old rat with irregular ventricular activation.
Differences in AT and AF induction between old and young
rats were signiŽ cant (P , 0.001, Chi-square test).
Effects of heptanol
The effects of increasing concentrations of heptanol (2 to
10 mM) on induction of AT/AF were evaluated in both
groups. The effects of increasing heptanol concentrations on
the inducibility of AT/AF were found to be biphasic in both
groups. At 2 to 5 mM, heptanol promoted inducible sus-
tained AT (.30 sec) in all Ž ve young rats studied (Figs. 4A
and 4B). However, when the heptanol concentration was
increased to 10 mM, AT could be induced in only 2 of 5
young rats (Fig. 4C). AF could not be induced in the any of
the Ž ve young rats at any of the heptanol concentrations (2
to 10 mM) tested. The mean CL of the AT in the young rats
Figure 4. Effects of burst atrial pacing on atrial rhythm in the presence of
did not change as heptanol concentration increased from 2
increasing heptanol concentrations (2 to 10 mM) in a young rat. Burst
to 5 mM and then to 10 mM (71.5 6 20.2 msec, 69.8 6 19.3
pacing induces sustained atrial tachycardia (AT) at (A) 2 mM and (B) 5
msec, and 55.7 6 8.9 msec, respectively). Furthermore, the mM heptanol. (C) No arrhythmia can be induced when the heptanol
CLs of the induced AT in the old rats at baseline and during concentration is raised to 10 mM. (D) Burst atrial pacing in the presence
perfusion of heptanol 2 mM (83.8 6 11.8 msec and 84.8 6 of 2 mM heptanol in an old rat induces sustained AF. However, increasing
20.4 msec) were not signiŽ cantly different from the CL of heptanol to (E) 5 mM and (F) 10 mM fails to induce arrhythmia. Abbre-
the AT induced in the young rats. In the old rats, when viations as in Figure 3.
 Hayashi et al. Aging and Atrial Fibrillation 805

Figure 5. Snapshots of optical maps during induced (A) atrial tachycardia (AT) and (C) atrial Ž brillation (AF) in an old rat. (B) Sites of three pixels (dots
numbered 1 to 3) from which optical action potentials are shown under panels A and B. During AT, a single wavefront propagates periodically from the
right to the left side (white arrow). During AT, the optical action potentials at sites 1, 2, and 3 show sequential activation from 1 to 3. Cycle length of AT
was 76 ms. During AF, multiple independent wavefronts were present and propagated in different directions (C; arrows). Optical action potentials and
right atrial bipolar electrogram (Beg) during AF show irregular (aperiodic) activity with a mean cycle length of 30 ms.

Figure 6. (A) Histologic Ž ndings in atrial tissues from a representative young rat (upper sections) and an old rat (middle sections) (hematoxylin and eosin,
original magniŽ cation 3400). (B) Histograms of mean atrial myocyte diameter. At all atrial sites, the mean atrial myocyte diameter was signiŽ cantly larger
in old rats than in young rats. (C) Mean of standard deviation of myocyte diameters at different atrial sites in both groups. LAA 5 left atrial appendage;
LAFW 5 left atrial free wall; RAA 5 right atrial appendage; RAFW 5 right atrial free wall.
806 Journal of Cardiovascular Electrophysiology Vol. 13, No. 8, August 2002
Figure 7. (A) Representative atrial tissue sections from a young rat (upper panels) and an old rat (lower panels) (trichrome stain, original magniŽ cation
3400). Note the signiŽ cant nonuniform increase in interstitial Ž brosis in the old atrium compared with the young atrium. (B) Histograms of mean
interstitial Ž brosis in both atria of young and old rats. (C) Mean of the standard deviation at different atrial sites in both groups. Abbreviations as in
Figure 6.
the young rats also showed a single large wavefront that
periodically propagated in the Ž eld of view. During AF,
multiple independent wavefronts were present, propagating
in different directions (Fig. 5C).
Histologic Ž ndings
Atrial myocyte size: Atrial myocardial cell size was sig-
niŽ cantly (P , 0.05) smaller in the young rats (4.1 6 0.6
mm) than in the old rats (5.5 6 0.7 mm) at all atrial sites
measured (Fig. 6). The moderate increase in atrial cell size
(width at the level of the nucleus in longitudinal sections)
was observed in both the RA and the LA (Fig. 6).
Interstitial atrial Ž brosis: A signiŽ cant increase in inter-
stitial atrial Ž brosis was present in old rats compared with
young rats. Figure 7 shows representative atrial sections in
an old and in a young atrium. The ratio of the area occupied
by interstitial Ž brosis to the total area was signiŽ cantly (P ,
0.01) higher in old rats than in young rats at all atrial sites
sampled in both chambers (15.5 6 14.9 vs 1.4 6 0.4, P ,
0.01) (Fig. 7). Increased Ž brosis was highly heterogeneous
in both atria, as indicated by the very high standard devia-
tions at all atrial sites (Fig. 7).
Discussion
Major Findings
Old rats were found to be associated with a signiŽ cantly
increased incidence of inducible AF. No AF could be in-
duced in the young rats. Partial cellular uncoupling with
heptanol in the young rats did not mimic the increased
incidence of inducible AF seen in the old rats. Another
Ž nding in this study was the biphasic changes of inducible
AT (young rats) and inducible AT and AF (old rats) during
increasing concentrations of heptanol perfusion. Whereas
lower concentrations of heptanol promoted inducible AT,
higher concentrations prevented inducible AT in the young
rats and AT/AF in the old rats.
Mechanism(s) of Aging-Associated Increase of AF
Vulnerability
The mechanism(s) of increased incidence of AF with
aging remains elusive.4 Changes in atrial tissue structure
appear to be of major importance in providing a substrate
for AF.1 2 - 1 4 In the present study, we found signiŽ cant Ž bro-
sis in the aged atria. The increased Ž brous tissue was
evident between atrial muscle Ž bers and atrial bundles, a
property that enhances nonuniform anisotropy and promotes
partial electrical uncoupling.5 In such a scenario, impulse
propagation might fail in the direction of decreased cellular
coupling but not in the more effectively coupled direc-
tion,1 5 ,1 6 causing conduction block, reentry, and AT. Fur-
thermore, cellular uncoupling might promote spatial disper-
sion of repolarization, which in turn may lead to
unidirectional conduction block and reentry.1 1 ,1 7 , 1 8 The abil-
ity of partial cellular uncoupling with heptanol to induce AT
with a single periodic activation wavefront in the young rats
supports this contention. However, partial cellular uncou-
pling failed to promote AF in the young rats. This observa-
tion implies that a generalized and uniform cellular uncou-

pling induced by perfusion of a cellular uncoupler does not
mimic nonuniform interstitial atrial Ž brosis as seen in the
aged atria. It is suggested that the increased incidence of
inducible AF in the old rats may be caused by the presence
of heterogeneous cellular uncoupling, a contention based on
the fact that uniform cellular uncoupling with perfusion of
heptanol1 9 , 2 0 failed to promote inducible AF in the young
rats. Furthermore, the Ž nding that heptanol only modestly
(i.e., from 3 to 4 of 6 rats) increased the incidence of
inducible AF in the old rats (complete elimination of induc-
ible AF resulted in all 6 old rats with heptanol .5 mM)
supports the importance of nonuniformity and heterogeneity
of uncoupling in promoting AF in the rat model. Complete
suppression of inducible AT and AF in both groups with
high heptanol concentrations (10 mM) is consistent with
block of wavefront propagation seen in the present study
and with previous studies in isolated pairs of cardiac myo-
cytes.1 7
Fibrosis, Heterogeneity, and AF
Preexisting heterogeneity is a major mechanism of wave-
break that converts tachycardia (single reentrant wavefront)
to Ž brillation (multiple wavefronts).2 1 It has been shown
that in dogs with induced heart failure, increased AF vul-
nerability was associated with increased atrial interstitial
Ž brosis and nonuniform excitability.2 2 The increased inci-
dence of inducible AF in the old rats in which AT could be
readily induced might result from the breakup of the single
activation wavefront during AT to multiple wavefronts, due
to the presence of preexisting nonuniformity in interstitial
Ž brosis in the aged atria. Another factor that may alter
anisotropy and promote AF in the aged atria is hypertrophy
of atrial myocytes. Aged atria showed about 20% increase
in cell diameter, a phenomenon that may slow and alter
anisotropic conduction2 3 and increase gap junctional resis-
tance2 4 that might facilitate inducible tachyarrhythmias.2 5 It
is conceivable that atrial cell hypertrophy may contribute to
increased AF in aged atria. We are less certain of the role of
atrial dynamic instability (restitution)2 1 in wavebreak of the
atrial activation wavefront. Provided that similarities of
atrial refractoriness between the young and the old rats
re ect similar recovery kinetics, the restitution hypothesis
of activation wavefront breakup may be of secondary im-
portance. We recently showed that heptanol 2 mM does not
alter transmembrane atrial action potential duration restitu-
tion kinetics or atrial refractoriness in canine atria.2 6 It
appears that the increased heterogeneous Ž brosis and non-
uniform atrial cellular uncoupling provide a suitable sub-
strate for activation wavefront breakups, leading to in-
creased incidence of pacing-induced AF in the aged atria.
More work is needed to determine the relative role of atrial
electrical restitution in promoting wavebreaks and AF in the
aged atria during rapid pacing.
Aging and P Wave Duration
Longer IACTs shown in the old rats could have been
caused by increased interstitial Ž brosis and cell hypertro-
phy, resulting in delayed total atrial depolarization time and
longer P wave duration. Reduced atrial conduction velocity
often is associated with increased AF vulnerability.7 ,2 7 , 2 8
Increased IACT associated with atrial interstitial Ž brosis in
humans may be responsible for the observed increased P
Hayashi et al. Aging and Atrial Fibrillation 807
wave duration and enhanced atrial vulnerability to arrhyth-
mias.2 9 - 3 2
Limitation of the Model
It is important to realize that results observed in the rat
model using the Langendorff setting may not mimic the
events seen in intact in situ aged human atria. Therefore,
one should be cautious when extrapolating the results of the
present study to clinical settings. Nevertheless, the in-
creased heterogeneous Ž brosis seen in aged human atria5
suggests that Ž brosis might contribute to aging-related AF
in man.
Clinical Implication
Heterogeneous interstitial Ž brosis might promote wave-
break and maintain AF once it is induced. Should similar
activation wavefront dynamics also exist in the aged human
atria,3 3 - 3 5 then the target for anti-AF drug therapy should
focus on eliminating the AF induction (trigger) factor(s) that
promotes launching of a rapid succession of activation
wavefronts. In anatomically remodeled and aged atria with
Ž brosis, rapid rates of activation promote continuous wave-
front breakups leading to AF maintenance.
Acknowledgments: The authors thank Drs. C. Thomas Peter and Prediman
K. Shah for support; Nina Wang for reading the manuscript; and Elaine
Lebowitz and Avile McCullen for secretarial and technical assistance.
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