the effect of genetic variability on coq levels ashley dudley
TRANSCRIPT
The Effect of Genetic
Variability on CoQ Levels
Ashley Dudley
Background• What is CoQ?
o Essential component of the mitochondrial electron transport chain, functioning as a transporter of electrons from several electron carriers
o Low CoQ levels are associated with cardiomyopathies, aging, and statin-induced myopathies
• CD36 is required in BAT for CoQ homeostasis and uptake in C57BL mice.o CoQ activates transcription of Ucp1.o BAT lacking CD36 was significantly larger and paler in color compared
to BAT from WT mice
Are there any genetic variants that can affect CoQ levels in mice?
Coenzyme Q
Methods• Mice
o Received from the Vulpe lab• Fed an iron sufficient diet• Fasted the night before
o All adult maleso 48 mice totalo Strains – very different genetically
• A/J• AKR/J• BALB/cJ• C3H/HeJ• C57BL/6J• DBA/2J
• Protocolo Coenzyme Q extraction (basic lipid extraction)o Protein quantification by BCA.o HPLC to quantify CoQ levels.
HPLC Chromatogram
Gastroc
BALB/cJ n=8
AKR/J n=8
DBA/2J n=8
A/J n=8 C3H/HeJ n=8
C57BL/6J n=8
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
CoQ9
CoQ
9 (
pm
ol/
ug p
rote
in)
*P=0.0140
AKR/J n=8
BALB/cJ n=8
DBA/2J n=8
A/J n=8 C3H/HeJ n=8
C57BL/6J n=8
00.020.040.060.080.1
0.120.140.160.180.2
CoQ10
CoQ
10 (
pm
ol/
ug p
rote
in)
**P=0.0015
BALB/cJ n=8
AKR/J n=8
DBA/2J n=8
A/J n=8 C3H/HeJ n=8
C57BL/6J n=8
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
Total CoQ
Tota
l C
oQ
10 (
pm
ol/
ug p
rote
in)
*P=0.0088
BAT
C3H/HeJ n=7
BALB/cJ n=8
A/J n=8 C57BL/6J n=7
DBA/2J n=8
AKR/J n=8
00.5
11.5
22.5
33.5
44.5
5
CoQ9
CoQ
9 (
pm
ol/
ug p
rote
in)
*P=0.0165
DBA/2J n=8
C3H/HeJ n=7
C57BL/6J n=7
BALB/cJ n=8
A/J n=8 AKR/J n=8
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
CoQ10
CoQ
10 (
pm
ol/
ug p
rote
in)
***P=0.0002
C3H/HeJ n=7
DBA/2J n=8
A/J n=8 C57BL/6J n=7
BALB/cJ n=8
AKR/J n=8
0
1
2
3
4
5
6
Total CoQ
Tota
l C
oQ
(pm
ol/
ug p
rote
in)
*P=0.0077
Liver
AKR/J n=8
C3H/HeJ n=8
AJ n=8 C57BL/6J n=8
BALB/cJ n=8
DBA/2J n=8
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
1.8
CoQ9
CoQ
9 (
pm
ol/
ug p
rote
in)
*P=0.0280
AJ n=8 C57BL/6J n=8
BALB/cJ n=8
AKR/J n=8
C3H/HeJ n=8
DBA/2J n=8
0.000
0.050
0.100
0.150
0.200
0.250
CoQ10
CoQ
10 (
pm
ol/
ug p
rote
in)
***P=0.0001
AJ n=8 C57BL/6J n=8
AKR/J n=8
C3H/HeJ n=8
BALB/cJ n=8
DBA/2J n=8
0
0.20.4
0.60.8
1
1.21.4
1.61.8
2
Total CoQ
Tota
l C
oQ
(pm
ol/
ug p
rote
in)
*P=0.0091
Serum
AKR/J n=6
A/J n=7 C57BL/6J n=7
BALB/cJ n=8
DBA/2J n=5
C3H/HeJ n=7
0.00
0.01
0.02
0.03
0.04
0.05
0.06
CoQ9
CoQ
9 (
pm
ol/
ul
seru
m)
**P=0.0034
A/J n=7 DBA/2J n=5
BALB/cJ n=8
C3H/HeJ n=7
C57BL/6J n=7
AKR/J n=6
-0.008
-0.006
-0.004
-0.002
0.000
0.002
0.004
0.006
0.008
0.010
CoQ10
CoQ
10 (
pm
ol/
ug p
rote
in)
**P=0.0038
A/J n=7 AKR/J n=6
BALB/cJ n=8
C57BL/6J n=7
DBA/2J n=5
C3H/HeJ n=7
0.00
0.01
0.02
0.03
0.04
0.05
0.06
Total CoQ
Tota
l C
oQ
(pm
ol/
ul
seru
m)
**P=0.0049
**p=0.0049
*p=0.0091 *p=0.0088
*p=0.0077
Comparison of Total CoQ
How Different are these Strains?
Mouse CharacteristicsC3H/HeJ AKR/J
• Blind by weaning age• Spontaneously develop
hair loss• Do not become obese
and fail to develop atherosclerosis when fed a high fat diet
• Highly susceptible to infection by Gram-negative bacteria
• High incidence of lymphatic leukemia (60-90%)
• Relatively resistant to diet-Induced Atherosclerosis
• Susceptible to audiogenic seizures
Future projects• Reconfirm CoQ levels with mice on a normal diet.• Examine CD36 levels in AKR/J vs C3H/HeJ strains
in BAT• Investigate CoQ levels in strains of mice more
similar to AKR and C3Ho Help hone in on loci that could be affecting CoQ levelso Then figure out which of those loci/genes is controlling CoQ levels
Acknowledgments• Andreas Stahl• Courtney Anderson• Angela Van