Patch clamp-assisted single neuron lipidomics Collin B. Merrill, Abdul Basit, Andrea Armirotti ...

49 Pages·2017·16.73 MB·English

Patch clamp-assisted single neuron lipidomics
Collin B. Merrill, Abdul Basit, Andrea Armirotti,Yousheng Jia,
Christine M. Gall, Gary Lynch, Daniele Piomelli
SUPPLEMENTARY FIGURES
16.70
a 100
m/z 734.56
e
c
n
a
d
n
u
b %
a
e
v
ati
el
R
0 Time
8.00 12.00 16.00 20.00 24.00
184.07
100
b
loss of FA 16:0-H2O
ce oup 496.33 [M+H]+
an dgr
d a 734.56
n e
u H loss of choline
ab % ne 478.32 Headgroup
e oli loss of FA 16:0 551.51
v h
ati C
el 460 500 540 580
R
0 m/z
200 400 600 800 1000
16.72
100
c
m/z 734.56
e
c
n
a
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ab %
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0 Time
8.00 12.00 16.00 20.00 24.00
Supplementary Figure S1. Identification of PC 32:0 (m/z = 734.56)
in diluted extract (1:50,000) of whole hippocampal tissue (A:
representative nLC tracing; B: mass spectrum) and isopropanol
extract of a single DG granule cell (C: representative nLC tracing).
Black tracings: neurons; red tracings: artificial cerebrospinal fluid.
18.33
a 100
m/z 762.60
e
c
n
a
d
n
u
b %
a
e
v
ati
el
R
0 Time
8.00 12.00 16.00 20.00 24.00
loss of
184.07
b 100 FA 16:0-H2O [M+H]+
524.36 762.59
loss of
e
nc oup loss of FA 18:0-H2O
nda d gr FA 1487:08.33 loss of loss of choline
e abu % ne Hea 496.33F5A0 61.63:50 He5a7d9 .g5r3oup
ativ Choli m/z
el 480 500 520 540 560 580
R
0 m/z
100 200 300 400 500 600 700 800
c 100 18.33
m/z 762.60
e
c
n
a
d
n
u
b %
a
e
v
ati
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0 Time
8.00 12.00 16.00 20.00 24.00
Supplementary Figure S2. Identification of PC 34:0 (m/z = 762.60)
in diluted extract (1:50,000) of whole hippocampal tissue (A:
representative nLC tracing; B: mass spectrum) and isopropanol
extract of a single DG granule cell (C: representative nLC tracing).
Black tracings: neurons; red tracings: artificial cerebrospinal fluid.
16.88
a 100
e m/z 760.58
c
n
a
d
n
u
ab %
e
v
ati
el
R
0 Time
8.00 12.00 16.00 20.00 24.00
184.07
b 100
[M+H]+
loss of 760.58
e FA 18:1-H2O
c p
dan grou 496.34 FA l1o6s:s0 o-Hf 2O
ative abun % Choline Head lFo4As7 s81 .8o3:2f1 5lFo0As4 s.13 64o:f50 22.35 577.l5oH1ses aodf cghrooulinpe
el
R m/z
460 500 540 580
0 m/z
100 200 300 400 500 600 700 800 900
c 100 16.89
m/z 760.58
e
c
n
a
d
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b
a %
e
v
ati
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R
0 Time
8.00 12.00 16.00 20.00 24.00
Supplementary Figure S3. Identification of PC 34:1 (m/z = 760.58)
in diluted extract (1:50,000) of whole hippocampal tissue (A:
representative nLC tracing; B: mass spectrum) and isopropanol
extract of a single DG granule cell (C: representative nLC tracing).
Black tracings: neurons; red tracings: artificial cerebrospinal fluid.
15.66
a 100
m/z 758.56
e
c
n
a
d
n
u
b %
a
e
v
ati
el
R
0 Time
8.00 12.00 16.00 20.00 24.00
184.07
b 100 loss of
[M+H]+
FA 16:0-H2O
758.56
loss of
loss of 520.34
e FA 16:0
c FA 18:2-H2O
an oup 502.33
und adgr loss of 494.31
b % e FA 18:2
a H
ative holine 476.31 m/z
el C 455 475 495 515 535
R
0 m/z
200 400 600 800 1000
15.65
c 100
m/z 758.56
e
c
n
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b %
a
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v
ati
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R
0 Time
6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00
Supplementary Figure S4. Identification of PC 34:2 (m/z = 758.56)
in diluted extract (1:50,000) of whole hippocampal tissue (A:
representative nLC tracing; B: mass spectrum) and isopropanol
extract of a single DG granule cell (C: representative nLC tracing).
Black tracings: neurons; red tracings: artificial cerebrospinal fluid.
100
m/z 790.63
a
e
c
n
a
d
n
u
b
a %
e
v
ati
el
R
19.82
0 Time
8.00 12.00 16.00 20.00 24.00
184.07
b 100
[M+H]+
e
nc oup 790.63
nda d gr
e abu % ne Hea
ativ Choli
el
R
0 m/z
100 200 300 400 500 600 700 800 900
c 100
m/z 790.63
e
c
n
a
d
n
u
b %
a
e
v
ati 19.82
el
R
0 Time
8.00 12.00 16.00 20.00 24.00
Supplementary Figure S5. Identification of PC 36:0 (m/z = 790.63)
in diluted extract (1:50,000) of whole hippocampal tissue (A:
representative nLC tracing; B: mass spectrum) and isopropanol
extract of a single DG granule cell (C: representative nLC tracing).
Black tracings: neu-rons; red tracings: artificial cerebrospinal fluid.
18.41
a 100
m/z 788.61
e
c
n
a
d
n
u
b
a %
e
v
ati
el
R
0 Time
6.00 10.00 14.00 18.00 22.00
[M+H]+
b 788.61
100
e 184.07 605.55
c
n
a loss of choline
d loss of
n Head group
bu up loss of FA 18:1-H2O
ative a % Head gro FA 18:5004.31524.35 m/z
el e 500 540 580 620
R n
oli
h
C
0
100 200 300 400 500 600 700 800 900 1000
c 100 18.39
m/z 788.61
e
c
n
a
d
n
u
b
a %
e
v
ati
el
R
0 Time
8.00 12.00 16.00 20.00 24.00
Supplementary Figure S6. Identification of PC 36:1 (m/z = 788.61)
in diluted extract (1:50,000) of whole hippocampal tissue (A:
representative nLC tracing; B: mass spectrum) and isopropanol
extract of a single DG granule cell (C: representative nLC tracing).
Black tracings: neurons; red tracings: artificial cerebrospinal fluid.
16.98
a 100
m/z 786.60
e
c
n
a
d
n
u
b %
a
e
v
ati
el
R
0 Time
8.00 12.00 16.00 20.00 24.00
[M+H]+
786.60
b 100
184.07
e 603.53
c
n loss of choline
a loss of
nd oup FA 18:1-H2O Head group
ve abu % Head gr lFoAs s1 8o5:f10 4.34522.35
elati oline m/z
R h 500 540 580 620
C
0 m/z
100 200 300 400 500 550 600 700 800
c 100 16.99
m/z 786.60
e
c
n
a
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ab %
e
v
ati
el
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0 Time
8.00 12.00 16.00 20.00 24.00
Supplementary Figure S7. Identification of PC 36:2 (m/z = 786.60)
in diluted extract (1:50,000) of whole hippocampal tissue (A:
representative nLC tracing; B: mass spectrum) and isopropanol
extract of a single DG granule cell (C: representative nLC tracing).
Black tracings: neurons; red tracings: artificial cerebrospinal fluid.
15.44
a 100
m/z 782.56
e
c
n
a
d
n
u
b
a %
e
v
ati
el
R
0 Time
8.00 12.00 16.00 20.00 24.00
[M+H]+
782.56
b 100 184.07
loss of
FA 20:4-H2O
e
ndanc group loss of 496.3l3oss of lFoAs s1 6o:f0 -H2O
e abu % e Head F4A7 280.3:42 F5A2 61.63:20 544.33 599.49 lHoessa do fg crohuopline
v n
ati oli
el Ch m/z
R 480 520 560 600
0 m/z
100 200 300 400 500 600 700 750 800
c 100 15.42
m/z 782.56
e
c
n
a
d
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b
a %
e
v
ati
el
R
0 Time
8.00 12.00 16.00 20.00 24.00
Supplementary Figure S8. Identification of PC 36:4 (m/z = 782.56)
in diluted extract (1:50,000) of whole hippocampal tissue (A:
representative nLC tracing; B: mass spectrum) and isopropanol
extract of a single DG granule cell (C: representative nLC tracing).
Black tracings: neurons; red tracings: artificial cerebrospinal fluid.
19.81
100
a
m/z 816.64
e
c
n
a
d
n
u
b
a %
e
v
ati
el
R
0 Time
8.00 12.00 16.00 20.00 24.00
[M+H]+
816.64
b 100
e 184.07
c
n
a
d p
bun % grou
a d
e ea
v H
ati ne
el oli
R h
C
0 m/z
100 200 300 400 500 600 700 800 900
c 100
m/z 816.64 19.83
e
c
n
a
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b %
a
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v
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0 Time
8.00 12.00 16.00 20.00 24.00
Supplementary Figure S9. Identification of PC 38:1 (m/z = 816.64)
in diluted extract (1:50,000) of whole hippocampal tissue (A:
representative nLC tracing; B: mass spectrum) and isopropanol
extract of a single DG granule cell (C: representative nLC tracing).
Black tracings: neurons; red tracings: artificial cerebrospinal fluid.