Transcriptional coactivators PGC-1?
and PGC-l? control overlapping
programs required for perinatal
maturation of the heart
Ling Lai,1,2,7Teresa C. Leone,1,2,7Christoph Zechner,1,2Paul J. Schaeffer,1,2,3Sean M. Kelly,1,2
Daniel P. Flanagan,1,2Denis M. Medeiros,4Attila Kovacs,1,2and Daniel P. Kelly1,2,5,6,8
1Center for Cardiovascular Research, Washington University School of Medicine, St Louis, Missouri 63110, USA;
2Department of Medicine, Washington University School of Medicine, St Louis, Missouri 63110, USA;3Department
of Zoology, Miami University, Oxford, Ohio 45056, USA;4Kansas State University, Manhattan, Kansas 66506, USA;
5Department of Molecular Biology and Pharmacology, Washington University School of Medicine, St Louis, Missouri 63110,
USA;6Department of Pediatrics, Washington University School of Medicine, St Louis, Missouri 63110, USA
Oxidative tissues such as heart undergo a dramatic perinatal mitochondrial biogenesis to meet the high-energy
demands after birth. PPAR? coactivator-1 (PGC-1) ? and ? have been implicated in the transcriptional control
of cellular energy metabolism. Mice with combined deficiency of PGC-1? and PGC-1? (PGC-1??−/−mice)
were generated to investigate the convergence of their functions in vivo. The phenotype of PGC-1?−/−mice
was minimal under nonstressed conditions, including normal heart function, similar to that of PGC-1?−/−
mice generated previously. In striking contrast to the singly deficient PGC-1 lines, PGC-1??−/−mice died
shortly after birth with small hearts, bradycardia, intermittent heart block, and a markedly reduced cardiac
output. Cardiac-specific ablation of the PGC-1? gene on a PGC-1?-deficient background phenocopied the
generalized PGC-1??−/−mice. The hearts of the PGC-1??−/−mice exhibited signatures of a maturational
defect including reduced growth, a late fetal arrest in mitochondrial biogenesis, and persistence of a fetal
pattern of gene expression. Brown adipose tissue (BAT) of PGC-1??−/−mice also exhibited a severe
abnormality in function and mitochondrial density. We conclude that PGC-1? and PGC-1? share roles that
collectively are necessary for the postnatal metabolic and functional maturation of heart and BAT.
[Keywords: Transcriptional regulation; heart development; mitochondria; energy metabolism]
Supplemental material is available at http://www.genesdev.org.
Received February 11, 2008; revised version accepted May 16, 2008.
The transcriptional coactivator peroxisome proliferator-
activated receptor ? (PPAR?) coactivator-1? (Ppargc1a, or
commonly called PGC-1?) was discovered based on its
functional interaction with the nuclear receptor PPAR?
in brown adipocytes (Puigserver et al. 1998). There-
after, two related transcriptional coactivators, PGC-1?
(Ppargc1b) and PGC-1-related coactivator (Pprc1, or com-
monly called PRC), were identified (Andersson and
Scarpulla 2001; Kressler et al. 2002; Lin et al. 2002a).
PGC-1? and PGC-1? exhibit the greatest degree of ho-
mology among the PGC-1 family members and are pref-
erentially expressed in tissues with high-capacity mito-
chondrial function such as heart, slow-twitch skeletal
muscle, and brown adipose tissue (BAT). PGC-1 coacti-
vators dock to specific target transcription factors, pro-
viding a platform for the recruitment of protein com-
plexes that exert powerful effects on target gene tran-
scription by remodeling chromatin and enabling access
by the RNA polymerase II machinery (Puigserver et al.
1999; Wallberg et al. 2003; Sano et al. 2007). Studies fo-
cused largely on PGC-1? have shown that it exerts its
biologic actions by coactivating a variety of nuclear re-
ceptor (e.g., PPAR?, PPAR?, estrogen-related receptor
[ERR] ?), and non-nuclear receptor (e.g., nuclear respira-
tory factors [NRFs], FOXO1) transcription-factor targets
(Wu et al. 1999; Vega et al. 2000; Huss et al. 2002; Puig-
server et al. 2003; Schreiber et al. 2003).
As opposed to the majority of known transcriptional
coactivators, the expression of PGC-1? and, to a lesser
extent PGC-1?, is highly inducible in response to devel-
opmental stage-specific and physiological cues (Puig-
server et al. 1998; Wu et al. 1999; Lehman et al. 2000;
7These authors contributed equally to this work.
E-MAIL email@example.com; FAX (407) 745-2001.
Article is online at http://www.genesdev.org/cgi/doi/10.1101/gad.1661708.
1948 GENES & DEVELOPMENT 22:1948–1961 © 2008 by Cold Spring Harbor Laboratory Press ISSN 0890-9369/08; www.genesdev.org
Finck and Kelly 2006; Handschin and Spiegelman 2006).
PGC-1? expression is induced by cold exposure, exer-
cise, and fasting in tissue-specific patterns. Studies of
murine heart have shown that PGC-1? is induced at
birth and during postnatal stages coincident with a dra-
matic energy metabolic maturation that involves a ro-
bust mitochondrial biogenic response and switch to re-
liance on fatty acids as the chief fuel (Lehman et al. 2000;
Lehman and Kelly 2002; Taha and Lopaschuk 2007; Bu-
roker et al. 2008). Overexpression studies in mammalian
cells in culture or in transgenic mice have shown that
both PGC-1? and PGC-1? are capable of activating the
expression of a cascade of genes involved in mitochon-
drial biogenesis and respiratory function in adipocytes,
cardiac myocytes, and myogenic cells (Wu et al. 1999;
Lehman et al. 2000; Lin et al. 2002a; St-Pierre et al.
2003). In addition, gain-of-function studies have shown
that PGC-1? activates gene regulatory programs in-
volved in hepatic gluconeogenesis, (Herzig et al. 2001;
Yoon et al. 2001; Puigserver and Spiegelman 2003; Koo et
al. 2004), muscle glucose uptake (Michael et al. 2001;
Wende et al. 2007), and slow-twitch muscle fiber type
determination (Lin et al. 2002b).
More recently, loss-of-function studies have been con-
ducted in mice in an attempt to define specific biological
roles for the PGC-1 coactivators and to determine the
necessity of these molecules in the regulation of biologic
processes in vivo. Two independent mouse lines with
generalized inactivation of the PGC-1? gene have been
generated (Lin et al. 2004; Leone et al. 2005). PGC-1?−/−
mice exhibit a surprisingly minimal phenotype under
basal physiologic conditions, indicating that it is dis-
pensable for the fundamental process of mitochondrial
biogenesis or fetal development. However, physiological
conditions that impose increased energy demands such
as cold exposure, fasting, or exercise, precipitate pheno-
types in the PGC-1?-deficient mice (Lin et al. 2004; Le-
one et al. 2005). The baseline cardiac phenotype of PGC-
1?-deficient mice is remarkably minimal given the im-
portance of a high-capacity mitochondrial system for
this organ. However, PGC-1?-deficient mice develop
ventricular dysfunction after prolonged pressure over-
load (Arany et al. 2006).
Recently, three independent lines of generalized PGC-
1?-deficient mice were generated. PGC-1?−/−mice ex-
hibit stress-induced phenotypes that are generally milder
but very similar to PGC-1?−/−mice (Lelliott et al. 2006;
Vianna et al. 2006; Sonoda et al. 2007). These results
suggest the possibility that PGC-1? and PGC-1? control
a subset of overlapping targets and are, therefore, capable
of compensating for the loss of the other factor in the
PGC-1 loss-of-function mice. To address this question
and to learn more about the role of PGC-1 coactivators in
vivo, we generated mice that are doubly deficient in
PGC-1? and PGC-1? (PGC-1??−/−) by targeting all four
alleles. As described by others, we found that our inde-
pendent line of PGC-1?-deficient mice exhibits a mild
basal phenotype similar to that of PGC-1?-deficient
mice. In striking contrast, PGC-1??−/−mice die shortly
after birth as a result of heart failure related to a perinatal
developmental arrest in cardiac maturation including a
block in mitochondrial biogenesis.
Generation and general characterization
of PGC-1?-null mice
Previously, three research groups have reported and char-
acterized PGC-1?-deficient mice (Lelliott et al. 2006; Vi-
anna et al. 2006; Sonoda et al. 2007). We generated an
independent line of PGC-1?-deficient mice. Briefly, a
cre-lox strategy was used to delete exons 4–6 of the mu-
rine PGC-1? gene (Supplemental Fig. 1A). The targeted
deletion introduced a predicted amino acid frameshift,
resulting in a premature stop codon in exon 7. The effi-
cacy of the gene targeting event and generation of the
mutant transcript was confirmed by PCR, Southern blot-
ting, RNA blotting, and immunoblotting studies (Sup-
plemental Fig. 1B–D).
Heterozygous mice (PGC-1?+/−) were bred to generate
PGC-1?−/−offspring. As described previously (Sonoda et
al. 2007), survival rates were modestly, but significantly
reduced (17% observed, 25% expected; Supplemental
Table 1). The surviving mice appeared normal. Given
that PGC-1? is necessary for adaptive physiological re-
sponses to stressors that demand increased mitochon-
drial oxidative capacity (Lin et al. 2004; Leone et al.
2005), the PGC-1?−/−mice were subjected to short-term
cold exposure and exercise. Six-week-old PGC-1?−/−
mice were subjected for 4 h to a cold environment (4°C)
without food. The PGC-1?−/−mice were unable to main-
tain core body temperature to the same degree as sex-
and weight-matched wild-type controls (Supplemental
Electron microscopic analysis of the BAT of PGC-
1?−/−mice did not reveal any significant abnormalities
in mitochondrial volume density or ultrastructure (data
not shown). Exercise performance was assessed using a
low-intensity, run-to-exhaustion exercise protocol on a
motorized treadmill. The mean running duration for the
PGC-1?−/−mice was less than that of PGC-1?+/+mice
(164 ± 14 vs. 202 ± 10 min, P < 0.05) (Supplemental Fig.
2B). Histologic analyses and electron microscopic stud-
ies of soleus muscles did not reveal any overt cellular or
ultrastructural abnormalities in the PGC-1?−/−mice
(data not shown). However, mean state 3 respiration
rates of mitochondria isolated from hindlimb muscle
were modestly but significantly decreased in PGC-1?−/−
mice compared with PGC-1?+/+controls (73.93 ± 6.12
vs. 102.58 ± 3.22 nmol O2per minute per milligram of
protein, P < 0.05) (Supplemental Fig. 2C). This finding of
reduced muscle state 3 respiratory rates in the PGC-
1?−/−mice is consistent with the results of recent studies
demonstrating that the expression of genes involved in
mitochondrial oxidative phosphorylation (OXPHOS) are
down-regulated in skeletal muscle (Lelliott et al. 2006;
Vianna et al. 2006; Sonoda et al. 2007) and BAT (Lelliott
et al. 2006; Sonoda et al. 2007) in PGC-1?-deficient
mouse lines. Taken together, these results demonstrate
PGC-1?/? deficiency causes cardiac death
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PGC-1?/? deficiency causes cardiac death
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