Current Biology, Volume
31
Supplemental Information
Neural mechanism of spatio-chromatic
opponency in the
Drosophila
amacrine neurons
Yan Li, Pei-Ju Chen, Tzu-Yang Lin, Chun-Yuan Ting, Pushpanathan Muthuirulan, Randall
Pursley, Marko Ili
c, Primo
z Pirih, Michael S. Drews, Kaushiki P. Menon, Kai G.
Zinn, Thomas Pohida, Alexander Borst, and Chi-Hon Lee
Figure
S
1
.
Light
stimulus
and
experimental
setup
.
Related
to
Figures
1
,
2
,
4
,
6
and
Figures
S
3
,
S
4
,
S
7
.
(A
–
B)
Emission
spectra
of
LEDs
used
in
the
multispectral
light
synthesizer
(B)
.
The
LED
synthesizer
was
used
to
produce
isoquantal
spectral
stimulation
over
a
wavelength
range
from
UV
to
red
by
using
15
LEDs,
a
grating,
an
aspheric
lens
and
a
light
guide
.
(C
–
D)
Emission
spectra
of
LEDs
used
in
two
-
color
light
stimuli
(D)
included
UV
(
375
nm)
and
blue
(
470
nm),
representing
UV
and
human
-
visible
light
stimuli
.
(E
–
F)
Emission
spectra
of
LEDs
used
in
the
custom
-
made
UV
array
(F)
for
mapping
the
spatio
-
temporal
receptive
fields
.
The
cylindrical
array
includes
360
UV
LEDs
(azimuth
×
elevation
:
24
×
15
)
with
maximal
spectrum
peaking
at
369
nm
.
Each
LED
covers
about
4
°
in
both
azimuth
and
elevation
.
(G
–
H)
Emission
spectra
of
LEDs
(G)
and
T
-
maze
set
up
(H)
used
in
the
phototactic
assay
.
Two
arms
of
the
T
-
maze,
fitted
with
transparent
polystyrene
vials,
were
illuminated
with
either
UV
(
373
nm)
or
green
(
523
nm)
LEDs
.
0
0.5
1
300
400
500
600
700
0
0.5
1
300
400
500
600
700
UV LED (375 nm)
Blue LED (470 nm)
Relative intensity (
a.u
.)
Relative intensity (
a.u
.)
optical fiber
lens
grating
E
F
H
G
A
C
B
D
360
373
389
402
423
449
468
486
548
569
584
609
625
645
674
(nm)
(nm)
0
0.5
1
300
400
500
600
700
0
0.5
1
300
400
500
600
700
Relative intensity (
a.u
.)
Wavelength (nm)
Relative intensity (
a.u
.)
UV array (369 nm)
UV LED (373 nm)
Green LED (523 nm)
Blue LED
(470 nm)
UV LED
(375 nm)
fiber
-
optic bundle
Green LED
(523 nm)
UV LED
(373 nm)
91.7 ̊
Fly
15 LED
56.5 ̊
24 LED
Figure
S
2
.
Immunostaining
showing
transgenes
expression
in
pale
and
yellow
Dm
8
of
flies
for
calcium
imaging
.
Related
to
Figures
1
–
2
.
Expression
of
syt
::
GCaMP
6
f
-
T
2
A
-
tdTomato
in
mixed
population
or
Dm
8
subtypes
using
different
drivers
or
manipulation
:
mixed
population
(A
–
D,
ort
C
2
b
-
Gal
4
,
corresponding
to
Figure
2
A),
pDm
8
(E
–
H,
ort
C
2
b
-
Gal
4
+
Dip
-
γ
-
Gal
80
,
corresponding
to
Figure
2
B),
yDm
8
(I
–
L,
ort
C
2
b
-
Gal
4
+
UAS
-
LDBD
+
Dip
-
γ
-
p
65
AD,
corresponding
to
Figure
2
C)
.
GCaMP
6
f,
tdTomato
and
photoreceptors
were
visualized
by
anti
-
GFP,
anti
-
RFP
and
anti
-
24
B
10
antibodies
(as
indicated)
.
Brackets
indicate
medulla
M
6
layer
.
Scale
bar
:
30
μm
.
GCaMP6f
tdTomato
α
24B10
Overlay
Mixed population
B
C
D
F
G
H
J
K
L
A
E
I
Yellow Dm8
Pale Dm8
Figure
S
3
.
ort
expression
in
Dm
8
s
restores
inhibitory
responses
to
UV
in
ort
mutant
flies
.
Related
to
Figure
2
.
B
A
ort rescue
Dm8>ort
(
ort
1
,ninaE
1
,HisCl1
134
,rh6
1
)
ort
mutant
(
ort
1
,ninaE
1
,HisCl1
134
,rh6
1
)
C
E
ort
mutant
(
ort
P306
/ort
1
,ninaE
1
,HisCl1
134
,rh6
1
)
ort rescue
Dm8>ort
(
ort
P306
/ort
1
,ninaE
1
,HisCl1
134
,rh6
1
)
D
w
+
,norpA
;Rh5
-
NorpA;
Dm8>GCaMP6f
ort
mutant
Dm8>
syt
::GCaMP6f
,
tdTomato
;
ort
1
,ninaE
1
,HisCl1
134
,rh6
1
ort rescue
Dm8>
syt
::GCaMP6f
,
tdTomato
, HA::ort;
ort
1
,ninaE
1
,HisCl1
134
,rh6
1
F
G
H
I
J
K
L
M
N
O
P
Q
R
S
T
U
ort
mutant
Dm8>
syt
::GCaMP6f
,
tdTomato
;
ort
P306
/ort
1
,ninaE
1
,HisCl1
134
,rh6
1
ort rescue
Dm8>
syt
::GCaMP6f
,
tdTomato
, HA::ort;
ort
P306
/ort
1
,ninaE
1
,HisCl1
134
,rh6
1
Constant
UV
Blue in UV
Constant
blue
UV in blue
tdTomato
GCaMP6f
α
HA
Overlay
(A
–
E)
Responses
of
Dm
8
s
to
UV
(
375
nm)
and
blue
(
470
nm)
two
-
color
light
stimuli
after
various
genetic
manipulations
.
Scale
bars
:
2
sec,
0
.
3
dF
/F
.
(A)
In
homozygous
ort
1
,ninaE
1
,hisCl
1
134
,rh
6
1
mutant
flies,
Dm
8
exhibits
light
-
on
excitation
.
(B)
Ort
expression
in
Dm
8
restored
inhibitory
responses
to
UV
in
homozygous
ort
1
,ninaE
1
,hisCl
1
134
,rh
6
1
mutant
flies
.
(C)
In
ort
P
306
/ort
1
,ninaE
1
,hisCl
1
134
,rh
6
1
mutant
flies,
Dm
8
exhibits
light
-
on
excitation
.
(D)
Ort
expression
in
Dm
8
restored
inhibitory
responses
to
UV
in
ort
P
306
/ort
1
,ninaE
1
,hisCl
1
134
,rh
6
1
mutant
flies
.
(E)
In
flies
with
only
functional
pR
8
photoreceptors
over
norpA
mutant
backgound
,
Dm
8
exhibits
light
-
on
excitation
.
(F
–
U)
Transgene
expression
in
flies
for
ort
-
rescue
experiments
(corresponding
to
Figures
S
3
A
–
S
3
D)
.
Two
types
of
mutant
background
were
tested
;
homozygous
ort
1
,ninaE
1
,hisCl
1
134
,rh
6
1
(F
–
M)
and
ort
P
306
/ort
1
,ninaE
1
,hisCl
1
134
,rh
6
1
(N
–
U)
.
By
using
anti
-
HA
antibody,
HA
::
ort
expression
in
Dm
8
was
detected
in
ort
-
rescue
groups
(L
and
T),
but
not
in
mutant
groups
(H
and
P)
.
Brackets
indicate
the
medulla
M
6
layer
.
Scale
bar
:
50
μm
.
Fitting Parameters
Neuron
R7
Dm8
Genetic
manipulation
wt
wt
NorpA
rescue
GluCl
α
RNAi
R7&R8
R1
–
R6&R7
R1
–
R6&R8
FWHM_center
(
°
)
6.71
8.58
8.96
9.35
9.48
8.88
FWHM_surround
(
°
)
―
59.99
51.46
59.99
36.59
―
Relative
amplitude
0
0.041
0.060
0.065
0.152
0
A
B
Figure
S
4
.
Characterization
of
spatial
properties
of
Dm
8
receptive
field
.
Related
to
Figure
4
.
(A)
GCaMP
6
f
signals
from
a
single
Dm
8
in
response
to
10
min
binary
white
noise
stimulus
.
(B)
Spatial
properties
of
Dm
8
and
R
7
RFs
under
various
manipulations
.
Parameters
derived
from
fitting
Difference
-
of
-
Gaussians
function
to
the
spatial
RF
.