Science 2005 Kujoth 481 4
Science 2005 Kujoth 481 4
Science 2005 Kujoth 481 4
Mammalian Aging
G. C. Kujoth et al.
Science 309, 481 (2005);
DOI: 10.1126/science.1112125
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REPORTS
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Finkbeiner, Nature 431, 805 (2004).
27. We thank A. Michael for support; S. Iyadurai and G.
Carlson for critical discussions; N. Nash, M. Sherman,
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0.75
0.50
0.00
F2-Isoprostanes
(ng/g)
1.8
Liver
0.25
WT
1.00
Heart
WT
D257A
0.4
Heart
Liver
0.6
0.3
0.2
0.3
0.1
0.0
D257A
0.0
WT
D257A
WT
D257A
D
3
Liver
Muscle (VL)
60
1.2
40
0.6
20
0.0
0
WT
D257A
12
Liver DNA
8-OHG/10 6 G
8-OHdG/10 6 dG
0
WT
D257A
Liver RNA
*
6
0
WT
D257A
WT
D257A
Fig. 2. Oxidative stress markers in isolated mitochondria and tissues from D257A mice. (A)
Hydrogen peroxide production was measured by a sensitive fluorometric assay in mitochondria
from wild-type (WT) and D257A mice at 9 months of age (N Q 8). (B) Protein carbonyl levels, a
marker of protein oxidation, were measured by an enzyme immunoassay in isolated mitochondria
from WT and D257A mice at 9 months of age (N Q 7). (C) F2-isoprostanes were measured by gas
chromatographynegative ion chemical ionization mass spectrometry in liver and skeletal muscle
(vastus lateralis) tissues from 6-month-old WT and D257A mice (N 0 6). (D) Oxidative damage to
DNA (8-OHdG) and RNA (8-OHG) was measured by high-performance liquid chromatography in
liver tissue of 9-month-old WT and D257A mice (N 0 9). *P G 0.05; error bars represent SEM.
Liver
Cleaved Caspase-3
(OD/mm2)
Cleaved Caspase-3
(OD/mm2)
482
200000
Testis
Cleaved Caspase-3
(OD/mm2)
80000
Muscle
*
60000
100000
40000
20000
5-mo 30-mo
Liver
3 mo
5-mo 30-mo
5-mo 30-mo
15000
Testis
3 mo
60000
10000
40000
5000
20000
Duod
3 mo
D257A
WT
100000
WT
Muscle
9 mo
WT
D257A
T hymus
3 mo
250000
D257A
Brain
9 mo
200000
75000
150000
50000
100000
25000
50000
WT
D257A
WT
D257A
WT
D257A
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tissues with the TUNEL (terminal deoxynucleotidyl transferasemediated deoxyuridine triphosphate nick end labeling) assay, which detects
apoptotic cells in situ. The 3-month-old D257A
mice showed significantly more TUNEL positive cells relative to wild-type mice in all tissues examined (Fig. 4). Together, these findings
strongly suggest that loss of critical, irreplaceable cells through apoptosis is a central mechanism of tissue dysfunction associated with the
accumulation of mtDNA mutations.
We have demonstrated that accelerated
development of aging phenotypes through
mtDNA mutations can occur in the absence of
increased ROS production or oxidative stress,
and that tissue dysfunction is likely to arise
through increased apoptosis. Moreover, we
have shown that increased caspase-3 activation
occurs in multiple tissues with normal aging.
Tissues that are composed of mitotic cells
display early caspase-3 activation in D257A
mice, whereas skeletal muscle displays a later
increase in cleaved caspase-3 and associated
tissue degeneration. It is also clear that some
cell types, such as spiral ganglion neurons, are
exquisitely sensitive to the effects of age-related
accumulation of mtDNA mutations. Because
D257A mice display high levels of mtDNA
mutations, it is possible that some of the
phenotypes in these animals may be due to
Fig. 4. Quantification of apoptosis by TUNEL in thymus (A), small intestine (B), and testis (C) of 3month-old WT (left panels) and D257A (center panels) mice. Arrowheads indicate TUNEL-positive
apoptotic nuclei. Numbers of apoptotic nuclei per 105 mm2 section (thymus), per 100 villi
(intestine), and per seminiferous tubule cross section (testis) were counted in hematoxylincounterstained sections from the indicated genotypes. Each bar represents apoptotic nuclei from
intestinal, thymus, and testis sections of at least four mice per genotype. *P G 0.05; error bars
represent SEM. Scale bar, 100 mm.
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(2001).
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J. M. Aiken, J. Gerontol. 48, B201 (1993).
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Myers, A. Aidoo, Mutat. Res. 526, 1 (2003).
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Chromatic Adaptation of
Photosynthetic Membranes
Simon Scheuring1* and James N. Sturgis2
Many biological membranes adapt in response to environmental conditions. We
investigated how the composition and architecture of photosynthetic membranes of a bacterium change in response to light, using atomic force microscopy. Despite large modifications in the membrane composition, the local
environment of core complexes remained unaltered, whereas specialized paracrystalline light-harvesting antenna domains grew under low-light conditions.
Thus, the protein mixture in the membrane shows eutectic behavior and can be
mimicked by a simple model. Such structural adaptation ensures efficient
photon capture under low-light conditions and prevents photodamage under
high-light conditions.
The atomic force microscope (1) is a powerful tool for imaging membrane proteins (2).
Recently, the first images at submolecular
resolution of native membranes have shed
light on the architecture of the photosynthetic
apparatus in different photosynthetic bacteria,
i.e., Blastochloris (Blc.) viridis (3), Rhodospirillum (Rsp.) photometricum (4, 5), Rhodobacter (Rb.) sphaeroides (6), and Rb.
blasticus (7). For photosynthesis to remain
efficient, the composition of the photosynthetic apparatus alters under different light
conditions. In many purple photosynthetic
1
Institut Curie, Unite Mixte de RechercheCNRS 168,
11 rue Pierre et Marie Curie, 75231 Paris Cedex 05,
Propre de Recherche9027 Laboratoire
France. 2Unite
dIndenierie de Systemes Macromoleculaires, Institut
de Biologie Structurale et Microbiologie, CNRS, 31
Chemin Joseph Aiguier, 13402 Marseille Cedex 20,
France.
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LH2, and (ii) paracrystalline peripheral antenna (LH2) domains. This two-phase structure could have an important functional role,
as the antenna fields may exclude quinone/
quinol (Q/QH2). This will markedly reduce
the membrane volume accessible to quinones and so accelerate transfer along preferential routes. As quinone diffusion is much
slower than light capture, energy transfer,
and the other electron transfer reactions (9),
this effect will increase efficiency. Our analysis further suggests a model for understanding the interactions between the different
components.
Qualitatively, we found the same photosynthetic complexes in high-light and lowlightadapted membranes. Comparison with
previous structural data for LH2 (5, 1015),
LH1-RC core complexes (3, 7, 1618), and
RCs (19, 20) allowed us to identify the small
rings (50 ) in diameter) as LH2 and the
large elliptical complexes as core complexes.
The LH1 assembly of the core complex forms
a closed ellipse of 16 LH1 subunits surrounding
an elongated RC (Fig. 1, A and B). The Rsp.
photometricum core complex is reminiscent of
that observed in Blc. viridis (3) and Rsp.
rubrum (16), monomeric and without a gap in
the LH1, unlike either the monomeric, Wcontaining core complex of Rps. palustris (18)
or the PufX-containing dimeric complexes of
Rb. sphaeroides (2123) and Rb. blasticus (7).
The functional consequences of the structural
variability of core complexes remain unclear
(24). The LH2 complexes were nonameric
rings (Fig. 1D). Rarely, LH2 that either lack
or have extra subunits are found (5). Such LH
complexes with extra subunits (Fig. 1C) were
found in both types of membranes and could
consist of LH1 or LH2 subunits or mixtures
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