10 min a b · 1 18 awake neuron Ð Ð /2 0 /2 respiratory phase (rad) Ð Ð /2 0 /2 respiratory...
TRANSCRIPT
10 min
1 s
Anesthesia
!"##
Water delivery port
Micropipette
Electrode holder
Head bar clamps
Odor tube
a b
c
Awake
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
GCL EPLGCL
Broadly tuned and respiration-independent inhibition in the olfactory bulb of awake miceBrittany N. Cazakoff, Billy Y. B. Lau, Kerensa L. Crump, Heike S. Demmer and Stephen D. Shea
Nature Neuroscience: doi:10.1038/nn.3669
a b
d
Rel
ativ
e vo
ltage
stra
in g
/ther
mo
cVo
ltage
/(dV/
dt)
1 432Time (s)
1 432Time (s)
c
1
0.5
Time lag (s)
Cor
rela
tion
–0.5 10.50–1
1
0.5
Time lag (s)
Cor
rela
tion
–0.5 10.50–1
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
Nature Neuroscience: doi:10.1038/nn.3669
a b
c d
0
0.5
1
1.5
2
2.5
3
Identified GC Putative GC
RM
I
0.2 0.4 0.60
1
2
3
4
5
Coupling strength (r)
Cou
nt
5 10 15 20 25 300
5
10
Baseline firing (spikes/s) C
ount
Ranked odors
4
2
0
–2
–4
Res
pons
e (s
pike
s/s)
n = 28n = 28
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
Nature Neuroscience: doi:10.1038/nn.3669
c
ba
Varia
nce
Varia
nce
*
Ranked odorsDuring
Ranked odorsOffset
Resp
onse
abo
ve b
asel
ine
(spi
kes/
s)
Resp
onse
abo
ve b
asel
ine
(spi
kes/
s)
0
5
10
15
20
25
30
35
Anesth Awake
Base
line
firin
g (s
pike
s/s) *
05
1015202530354045
0102030405060708090
50 100
Anesth AwakeDuring
Anesth AwakeOffset
*
-3
-2
-1
0
1
2
3
4
-3
-2
-1
0
1
2
3
4** *
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
Nature Neuroscience: doi:10.1038/nn.3669
ba
2
3
4
5
Still Run
Brea
thin
g fre
quen
cy (H
z)
6*
Still Run0
2
4
6
8
10
Spik
es/s
0 10 20 300
10
20
30
Stillrate (spikes/s)
Run
rate
(spi
kes/
s)c
Mov
eBr
eath
Spik
e
n = 33
n = 16
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?C@"D"B"6,(."3*&()E"@2%3("+%,"*)"F")"%&."G"25H)"04,"342454-*4&="F"5I"04,"+,(%-$*&'"%&."F!J"5I"04,"/#"):*K*&'!"
6+7"L,(%-$*&'"0,(M>(&2<"*&2,(%)()"1*-$",>&&*&'!"L,(%-$*&'".>,*&'"-$("(&-*,(",(24,.*&'")())*4&"1%)"+*&&(."*&-4"B")"
*&-(,;%3)="%&."%"5(%&",%-(".>,*&'":(,*4.)"40",()-"4,".>,*&'":(,*4.)"40",>&&*&'"1%)"2%32>3%-(.!"N4,"%33"%&*5%3)"6&"O"
FP"2(33)"0,45"Q"%&*5%3)7="+,(%-$*&'")*'&*0*2%&-3<"*&2,(%)(.".>,*&'",>&&*&'"6C(%&"+,(%-$*&'",%-("R")!(!5!",()-"O"
B!PS"R"T!TQE",>&&*&'"U!PQ"R"T!FPE"V6FJ7"O"FJW=""XY"O""T!TTTTBP="C%&&Z[$*-&(<"V7!"627"/#"%2-*;*-<"*)"3%,'(3<"
*&;%,*%&-"1*-$"%2-*;*-<!"C(%&"):*K(",%-("R")!(!5!"+(-1((&":(,*4.)"40",()-"6Q!FJ"R"F!GP"):*K()H)7"%&.":(,*4.)"40"
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[$*-&(<"V="3(0-7!""Y34--*&'"*&.*;*.>%3".%-%":4*&-)"65*..3(="3(0-"$*)-4',%5="&"O"UU"2(33)"0,45"FS"%&*5%3)7"*&.*2%-()"
%")>+Z:4:>3%-*4&"40"2(33)"*)"54.>3%-(."1*-$"%2-*;*-<!"\4--(.",(."3*&(",>&,%-("O")-*33,%-(!"
-1 0 1024681012
(Runrate-stillrate)(Runrate+stillrate)
Cou
nt Still > Run >Run Still
Nature Neuroscience: doi:10.1038/nn.3669
1
18
Neu
ron
Awake
– – /2 0 /2Respiratory phase (rad)
– – /2 0 /2Respiratory phase (rad)
3
2
1
Cou
nt
b
c
0.2 0.4 0.6 0.8
12
8
4
Cou
nt
Norm
alizedcount
1
0
Awake (n=18)
a
Coupling strength (r)
Inhalation Exhalation
Supplementary Figure 6. Assessment of respiratory coupling when GC spiking is aligned to breathing onset. (a)
Histogram (bin size = 0.1) of respiratory coupling strength (r) for awake state GCs. As with alignment to peak inspira-
tion, GCs still show weak respiratory coupling. (b) Heatmap depicting baseline respiratory phase spike histograms for all
that cell. White dots denote the mean phase angle of firing for each cell. A representative breath trace is shown in black
-
ness (gray).
Nature Neuroscience: doi:10.1038/nn.3669
1
1
0.8
0.6
0.4
0.2
432Quartile
Cou
plin
g st
regt
h (r)
SlowerFasterRespiratory rate
1
1
0.8
0.6
0.4
0.2
432Quartile
Cou
plin
g st
regt
h (r)
LongShortInspiration dur
1
1
0.8
0.6
0.4
0.2
432Quartile
Cou
plin
g st
regt
h (r)
LongShortExpiration dur
1
1
0.8
0.6
0.4
0.2
432Quartile
Cou
plin
g st
regt
h (r)
LargeSmallInspiration amp
a
dc
b
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?+$43-,#(!5*4!#$)'!)#,,!$))*46-./!3*!*.#!*5!5*+4!74#$3'!&$4$8#3#4(9!@#(&-4$3*4A!)*+&,-./!(34#./3'!5*4!(&-%#(!6-6!
.*3!(A(3#8$3-)$,,A!>$4A!0-3'!$.A!*5!3'#!74#$3'!&$4$8#3#4(!0#!8#$(+4#6B!!74#$3'-./!4$3#!C$DE!-.(&-4$3*4A!6+4$3-*.!
C7DE!#F&-4$3*4A!6+4$3-*.!C)DE!$.6!!-.(&-4$3*4A!$8&,-3+6#!C6D9!!2*8&$4-./!3'#!6-(34-7+3-*.!*5!)*+&,-./!(34#./3'!
$)4*((!?+$43-,#(!5*4!#$)'!&$4$8#3#4E!0#!($0!.*!(-/.-5-)$.3!6-55#4#.)#(!C.!G!;<!)#,,(H!I!G!J9KL!C$DE!I!G!J9MM!C7DE!I!G!J9KJ!C)DE!I!G!J9MM!C6DE!N4+(%$,O"$,,-(D9!
Nature Neuroscience: doi:10.1038/nn.3669
12
8
4
0.2 0.80.60.4 1.0
Cou
nt
Coupling strength (r)
n = 18
12
8
4
0.2 0.80.60.4 1.0
Cou
nt
Coupling strength (r)
n = 18
12
8
4
0.2 0.80.60.4 1.0
Cou
nt
Coupling strength (r)
n = 18
a
b
c
Baseline
All other sniffs
First sniff
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
Nature Neuroscience: doi:10.1038/nn.3669
baseline firing (spikes/s)
a
024681012
kx iso
b
5 10
5
0
10
coun
t
base
line
firin
g (s
pike
s/s)
–3–2–1
0123
ranked odors ranked odors
KX (n = 4)iso (n = 19)
during after
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
KX (n = 3)iso (n = 19)
Nature Neuroscience: doi:10.1038/nn.3669
Table 1. List of Odorants
* Denotes odors used in formulation of tuning curves
Monomolecular Odorants
Natural Odorants
acetophenone* ethyl benzoylacetate
3-methyl-2-buten-1-ol
anise oil
4-allyanisole
ethyl butyrate octanal*
cedarwood
2-butenol
ethyl propionate 1-pentanol* cinnamon oil
butyl formate*
eucalyptol
pentylacetate
cloves
butyl propionate*
eugenol*
2-phenylethanol
cumin
butyraldehyde
heptanal propionicaldehyde
nutmeg
camphor 2-heptanone
propyl acetate peanut butter
cineole*
4-heptanone pyrrolidine
peppermint
citral hexanal valeraldehyde
vanilla extract
cyclohexylacetate*
2-hexanone valeric acid*
decanolactone
limonene*
Supplementary
Nature Neuroscience: doi:10.1038/nn.3669