Sin ttulo de diapositiva

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From López-Torres et al. Mech Ageing Dev 70
(1993) Barja et al Free Radic Res 21 (1994)
Pérez-Campo et al J Comp Physiol B 163
(1994) Barja et al Comp Biochem Physiol Biochem
Mol Biol 108 (1994)
From other four different independent
laboratories
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(LONG-LIVED ANIMAL SPECIES)
(Barja cols. MAD 1993 CBP1994 JCP, 1994
Free Rad. Res. 94, Review (written in 1994,
passed through 4 journals finally published
in J. Comp. Physiol. In 1998
LOW ANTIOXIDANT LEVELS
(HYPOTHESIS) 1993
LOW RATE OF MITOCHONDRIAL OXYGEN RADICAL
PRODUCTION
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COMPARATIVE STUDIES OF MITOCHONDRIAL H2O2
PRODUCTION IN MAMMALS AND BIRDS WITH DIFFERENT
MAXIMUM LONGEVITIES (MLSP)
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PIGEON MLSP 35 YEARS
RAT MLSP 4 YEARS
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S
S
S
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Succinate
Cx II
Pyruvate/ Malate
TTFA
AA
O2
c
Cx IV
Cx III
Q
Cx I
ROT
H2O
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(LONG-LIVED ANIMAL SPECIES)
LOW RATE OF MITOCHONDRIAL OXYGEN RADICAL
PRODUCTION
(HYPOTHESIS)
LOW OXIDATIVE DAMAGE IN MITOCHONDRIAL DNA
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8-oxoGua
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r-0.92 plt0.000
(HEART)
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8
Mouse
(SAME RESULT IN BRAIN)
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8-oxodG/ 105dG in heart mt DNA
Guinea pig
Rat
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Rabbit
Pig
2
Sheep
Horse
Cow
0
10
40
30
20
50
MAXIMUM LONGEVITY (YEARS)
G.Barja A. Herrero. FASEB J. 14 312-318 (2000)
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2
(BRAIN)
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Horse
8-oxodG/ 105dG in nDNA
Mouse
Guinea Pig
Rat
Cow
Rabbit
0
10
40
30
20
50
0
MLSP (YEARS)
(SIMILAR RESULTS IN HEART)
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H2O2 PRODUCTION HEART MITOCHONDRIA
1.2
AC
OC
1.0
OR
0.8

nmoles H2O2/ min mg prot
0.6
0.4
0.2
0.0
Pyruvate/ malate Long-term Caloric Restriction (1
year)
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H2O2 PRODUCTION HEART MITOCHONDRIA
2.5
AC
OC
OR
2.0
1.5
nmoles H2O2/ min mg prot
1.0
0.5
0.0
Succinate ( Rotenone) Long-term Caloric
Restriction (1 year)
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Succinate
Cx II
Pyruvate/ Malate
TTFA
AA
O2
c
Cx IV
Cx III
Q
Cx I
ROT
H2O
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Cx I
NADH
1,2-NAPHTOQUINONE
p-ClHg BENZOATE
FERRICYANIDE
ETHOXYFORMIC ANHYDRIDE
ROTENONE
MENADIONE
Q
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O2 CONSUMPTION HEART MITOCHONDRIA
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AC
OC
OR
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nmoles O2/ min mg prot
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8
0
Pyruvate/ malate Long-term Caloric Restriction (1
year)
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FREE RADICAL LEAK HEART MITOCHONDRIA
2.5
AC
OC
2.0
OR
1.5

()
1.0
0.5
0.0
Pyruvate/ malate Long-term Caloric Restriction (1
year)
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Age of animals at time of analysis adult (7
months), old (24 months). Results are means SEM
(n). No significant differences due to age or
restriction were observed.
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8-oxodG in HEART mtDNA
Long-term Caloric Restriction (1 year)
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CALORIC RESTRICTION
LONG-LIVED ANIMALS
(-)
(-)
MITOCHONDRIAL ROS PRODUCTION RATE
MITOCHONDRIAL DNA
RATE OF ACCUMULATION OF mtDNA MUTATIONS
AGING RATE
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Survival plots of dwarf mice. Left panel Snell
dwarf mice and normal controls. Right panel Ames
dwarf mice and normal controls. Reproduced from
Bartke and Turyn 2001.
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5.0
WILD TYPE
AMES DWARF
4.0

3.0
8-oxodG/105dG in mtDNA

2.0
1.0
0.0
HEART
BRAIN
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Short-lived animals Ad libitum animals
HIGH 8-oxodG
High (8-oxo mtDNA repair ? (post.mitot.)
(High mitoch. ROS product.)
High attack
Long-lived animals Restricted animals
LOW 8-oxodG
Low 8-oxodG mtDNA repair ? (post.mitot.)
(Low mitoch. ROS product.)
Low repair ?
Low attack
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DOUBLE BOND INDEX OF FATTY ACIDS HEART
(MOUSE)
(Rat)
(Pig)
(G. pig)
(Horse)
(Rabbit)
(Cow)
(Sheep)
MAX. LONGEVITY (YEARS)
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Relationship between maximum longevity and
linoleic acid (182n-6) and docosahexanoic acid
(226n-3) contents in heart phospholipids of 8
mammalian species
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CALORIC RESTRICTION
LONG-LIVED ANIMALS
? DBI
(-)
(-)
MITOCHONDRIAL ROS PRODUCTION RATE
?
? MDA
MITOCHONDRIAL DNA
?
? Prot. ox.
RATE OF ACCUMULATION OF mtDNA MUTATIONS
AGING RATE
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OXYGEN RADICAL PRODUCTION IS NOT NECCESSARILY
PROPORTIONAL TO OXYGEN CONSUMPTION
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Low rate of electron flux
ROS
ROS
High rate of electron flux
ROS
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Positive fluorescent metod Specific for
H2O2 Sensitive Does not alter mitochondria Instant
aneous response to H2O2 No antioxidant
interference
(Barja G. (2002) J. Bioenerg. Biomembr. 33227-233
HOMOVANILLIC ACID
H2O2
HORSERADISH PEROXIDASE
DIMER
FLUORESCENCE 312 nm EXC 420nm EM
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(Endogenous generation of oxidative damage)
(DBI)
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THE RATE OF GENERATION OF ENDOGENOUS DAMAGE IS
THE CAUSE OF AGING
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COLLABORATION IS MUCH BETTER THAN COMPETITION
(A L S O I N S C I E N C E)
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