Cell, Volume
186
Supplemental information
Parallel neural pathways control sodium consumption
and taste valence
Yameng Zhang, Allan-Hermann Pool, Tongtong Wang, Lu Liu, Elin Kang, Bei Zhang, Liang
Ding, Kirsten Frieda, Richard Palmiter, and Yuki Oka
Condition
Comparison
Test
Post hoc test
Significance
p value
n
LS vs Water
***
0.0002
10, 10
LS vs KCl
***
0.0002
10, 20
LS vs HS
ns
0.9395
10, 21
LS vs LS + KCl
ns
>0.9999
10, 9
LS vs LS + CaCl
2
ns
0.3942
10, 9
LS vs LS + MgCl
2
ns
>0.9999
10, 8
LS vs LS + quinine
ns
>0.9999
10, 9
LS vs Water
ns
>0.9999
14, 7
LS vs KCl
***
<0.0001
14, 7
LS vs HS
***
<0.0001
14, 14
LS vs LS + KCl
***
<0.0001
14. 9
LS vs LS + CaCl
2
***
0.0005
14, 9
LS vs LS + MgCl
2
*
0.0103
14, 10
LS vs LS + quinine
*
0.0398
14, 10
LS vs Water
ns
>0.9999
13, 13
LS vs KCl
***
0.0005
13, 13
LS vs HS
**
0.0006
13, 13
LS vs LS + KCl
***
<0.0001
13, 12
LS vs LS + CaCl
2
***
0.0008
13, 6
LS vs LS + MgCl
2
*
0.0216
13, 6
LS vs LS + quinine
*
0.029
13, 6
Pdyn
Cre/wt
, LS
ns
0.882
9, 11
Pdyn
Cre/wt
, HS
ns
0.0529
15, 12
Pdyn
Cre/wt
, LS + KCl
***
<0.0001
12, 10
Pdyn
Cre/wt
, LS + quinine
***
<0.0001
12, 13
Ptger3
Cre/wt
, thirst, water
ns
0.9872
8, 8
Ptger3
Cre/wt
, thirst, 60 mM NaCl
ns
0.9901
4, 4
Ptger3
Cre/wt
, thirst, 250 mM NaCl
***
0.0001
7, 7
Ptger3
Cre/wt
, thirst, 500 mM NaCl
***
<0.0001
12, 12
Ptger3
Cre/wt
, thirst, LS + KCl
***
0.0003
6, 6
Ptger3
Cre/wt
, thirst, LS + quinine
***
0.0002
6, 6
Ptger3
Cre/wt
, thirst, water
ns
0.4747
7, 7
Ptger3
Cre/wt
, thirst, 500 mM NaCl
ns
0.9998
5, 5
Ptger3
Cre/wt
, -sodium, LS
ns
0.084
7, 7
Ptger3
Cre/wt
, -sodium, LS + KCl
*
0.0192
7, 7
Ptger3
Cre/wt
, -sodium, LS + quinine
***
0.0002
7, 7
Figure 4B
Pdyn
Cre/wt
, pre-LC ChR2 stim
Mann-Whitney test
NA
*
0.0286
4, 4
Figure 4C
Ptger3
Cre/wt
, SFO ChR2 stim
Mann-Whitney test
NA
**
0.0048
4, 6
Pdyn
Cre/wt
and WT, pre-LC caspase, SFO
Mann-Whitney test
NA
ns
0.9048
4, 5
Pdyn
Cre/wt
and WT, pre-LC caspase, pre-LC
Mann-Whitney test
NA
***
0.0007
6, 8
Ptger3
Cre/wt
and WT, SFO caspase, SFO
Mann-Whitney test
NA
*
0.0286
4, 4
Ptger3
Cre/wt
and WT, SFO caspase, pre-LC
Mann-Whitney test
NA
ns
0.6032
4, 5
Ptger3
Cre/wt
, thirst, water
ns
0.9504
8, 8
Ptger3
Cre/wt
, thirst, 0.125 mM quinine
***
<0.0001
5, 5
Ptger3
Cre/wt
, thirst, 0.25 mM quinine
***
0.001
8, 8
Ptger3
Cre/wt
, thirst, water
ns
0.9693
8, 8
Ptger3
Cre/wt
, thirst, 10 mM citric acid
**
0.003
11, 11
Ptger3
Cre/wt
, thirst, 20 mM citric acid
**
0.0021
11, 11
Ptger3
Cre/wt
, food deprivation, water
ns
0.9989
7, 7
Ptger3
Cre/wt
, food deprivation, 0.5 mM AceK
ns
0.9998
4, 4
Ptger3
Cre/wt
, food deprivation, 1 mM AceK
ns
0.9801
4, 4
Ptger3
Cre/wt
, food deprivation, 2 mM AceK
ns
0.8498
6, 6
Ptger3
Cre/wt
, thirst, 0 μM capsaicin
ns
0.296
4, 4
Ptger3
Cre/wt
, thirst, 0.3 μM capsaicin
ns
0.6279
5, 5
Ptger3
Cre/wt
, thirst, 1 μM capsaicin
ns
0.7295
5, 5
Ptger3
Cre/wt
, food deprivation, glucose
Pure sweet vs Sweet + Bitter
**
0.0044
10, 8
Ptger3
Cre/wt
, food deprivation, glucose + quinine
-Stim vs +Stim
*
0.0181
8, 9
Ptger3
Cre/wt
, food deprivation, AceK
Pure sweet vs Sweet + Bitter
**
0.0012
10, 9
Ptger3
Cre/wt
, food deprivation, AceK + quinine
-Stim vs +Stim
*
0.0437
9, 9
Ptger3
Cre/wt
, food deprivation, MPG + IMP
Pure umami vs Umami + Bitter
***
0.0002
7, 7
Ptger3
Cre/wt
, food deprivation, MPG + IMP + quinine
-Stim vs +Stim
*
0.0165
7, 8
-Sodium, SFO
*
0.0325
4, 5
-Sodium, pre-LC
ns
0.9975
7, 6
-Sodium, water
ns
>0.9999
4, 4
Figure 5B
-Stim vs +Stim
Figure 5C
-Stim vs +Stim
Figure 5D
-Stim vs +Stim
Figure 5G
Figure 6B
Ptger3
Cre/wt
vs
Ptger3
Cre/Cre
Figure 3F
Vch vs CNO
Vch vs CNO
SFO vs pre-LC
Figure 4D
Cre vs WT
Figure 4E
Cre vs WT
Figure 5A
-Stim vs +Stim
WT, -Sodium
WT, Thirst
WT, Sated
Figure 1A
+Stim vs -Sodium
Figure 1D
-Stim vs +Stim
Figure 3D
Figure 3E
-Stim vs +Stim
Two-way ANOVA
Sidak's multiple comparisons test
Sidak's multiple comparisons test
Sidak's multiple comparisons test
Dunn’s multiple comparison test
Dunn’s multiple comparison test
Dunn’s multiple comparison test
NA
Sidak's multiple comparisons test
Two-way ANOVA
Two-way ANOVA
One-way ANOVA (Kruskal–Wallis test)
One-way ANOVA (Kruskal–Wallis test)
One-way ANOVA (Kruskal–Wallis test)
Two-way ANOVA
Two-way ANOVA
Two-way ANOVA
Two-way ANOVA
Two-way ANOVA
One-way ANOVA (Kruskal–Wallis test)
One-way ANOVA (Kruskal–Wallis test)
One-way ANOVA (Kruskal–Wallis test)
Mann-Whitney test
Two-way ANOVA
Sidak's multiple comparisons test
Sidak's multiple comparisons test
Sidak's multiple comparisons test
Sidak's multiple comparisons test
Dunn’s multiple comparison test
Dunn’s multiple comparison test
Dunn’s multiple comparison test
Sidak's multiple comparisons test
Table S1. Statistics Table, related to Figures 1-7 and S1-7
-Sodium,LS
ns
0.999
5, 10
-Sodium, HS
*
0.0114
5, 6
-Sodium, LS + KCl
**
0.0065
5, 4
-
Sodium, LS + quinine
***
0.0004
4, 7
WT, -sodium, SFO
**
0.0016
5, 5
WT, -sodium, pre-LC
ns
0.9983
5, 5
WT, -sodium, water
ns
>0.9999
5, 5
WT, -sodium, LS
ns
0.7055
8, 11
WT, -sodium, HS
***
0.0003
13, 13
WT, -sodium, LS + KCl
*
0.0112
8, 8
WT, -sodium, LS + quinine
*
0.0256
7, 8
Sated vs -Sodium
*
0.0117
9, 8
Sated vs +Sodium
ns
0.9467
9, 6
Sated vs Formain 30 min
*
0.0435
9, 8
Sated vs Formain 90 min
ns
0.5024
9, 8
Figure 7B
WT, PGE2-AMCA dye
Veh vs PGE2-AMCA
Mann-Whitney test
NA
**
0.0033
7, 9
Figure 7C
Ptger3
Cre/wt
, photometry
Veh vs PGE2
Mann-Whitney test
NA
**
0.0012
7, 6
Ptger3
Cre/wt
, sated, 500 mM NaCl
ns
0.9996
4, 4
Ptger3
Cre/wt
, thirst, 500 mM NaCl
***
0.0004
5, 5
Ptger3
Cre/Cre
, sated, 500 mM NaCl
ns
0.9971
5, 5
Ptger3
Cre/Cre
,
thirst, 500 mM NaCl
ns
0.0562
4, 4
WT,
scramble
-shRNA, thirst, 500 mM NaCl
***
<0.0001
5, 5
WT,
shRNA
-shRNA, thirst, 500 mM NaCl
ns
0.3824
10,10
WT, -sodium, 60 mM NaCl
ns
0.1564
4, 4
WT, -sodium, 500 mM NaCl
**
0.0021
5, 5
Thirst vs -Sodium
**
0.0065
8, 9
Thirst vs Thirst + PGE2
*
0.0238
8, 8
Figure 7H
Ptger3
Cre/wt
, thirst, Dlight
Veh vs CNO
Wilcoxon test
NA
**
0.0078
8, 8
WT, Water
Mann-Whitney test
NA
***
0.0001
7, 10
WT, KCl
Mann-Whitney test
NA
ns
0.0568
7, 20
WT, LS
Mann-Whitney test
NA
ns
0.4031
14, 10
WT, HS
Mann-Whitney test
NA
***
<0.0001
14, 21
WT, LS + KCl
Mann-Whitney test
NA
***
<0.0001
9, 9
WT, LS + CaCl
2
Mann-Whitney test
NA
***
0.0005
9, 9
WT, LS + MgCl
2
Mann-Whitney test
NA
***
<0.0001
10, 8
WT, LS + quinine
Mann-Whitney test
NA
***
0.0004
10, 9
+Stim vs -Sodium
ns
>0.9999
9, 11
+Stim vs -Stim
*
0.0116
9, 4
-Stim vs -Sodium
*
0.0131
4, 11
+Stim vs -Sodium
ns
0.2651
15, 12
+Stim vs -Stim
*
0.0364
15, 4
-Stim vs -Sodium
***
0.001
4, 12
+Stim vs -Sodium
**
0.0033
12, 10
+Stim vs -Stim
ns
0.1552
12, 5
-Stim vs -Sodium
***
<0.0001
5, 10
+Stim vs -Sodium
***
0.0005
12, 13
+Stim vs -Stim
ns
>0.9999
12, 4
-Stim vs -Sodium
***
0.0009
4, 13
Figure S1E
Pdyn
Cre
, HS/LS
+Stim vs -Sodium
Mann-Whitney test
NA
*
0.0321
15, 12
Pdyn
Cre
, thirst, 500 mM NaCl
***
<0.0001
5, 5
Pdyn
Cre
, thirst, 500 mM NaCl with amiloride
ns
0.6519
6, 6
Figure S3D
Ptger3
Cre/wt
,
thirst, comtrol for ChR2
-Stim vs +Stim
Wilcoxon test
NA
ns
0.8125
5, 5
Ptger3
Cre/wt
, thirst, 500 mM NaCl
***
<0.0001
5, 5
Ptger3
Cre/wt
, thirst, 500 mM NaCl with amiloride
***
<0.0001
5, 5
Ptger3
Cre/wt
, thirst, 250 mM NaCl
*
0.0111
8, 9
Ptger3
Cre/wt
, thirst, 500 mM NaCl
**
0.0037
6, 12
Figure S3H
Ptger3
Cre/wt
,
hypovolemia , HS
Vch vs CNO
Wilcoxon test
NA
*
0.0313
6, 6
Ptger3
Cre/wt
, -sodium, LS
ns
0.1997
6, 6
Ptger3
Cre/wt
, -sodium, LS + KCl
**
0.0088
6, 6
Ptger3
Cre/wt
, -sodium, LS + quinine
***
0.0008
6, 6
WT and
Trpm5
-/-
, thirst, water
ns
0.8056
7, 3
WT and
Trpm5
-/-
, thirst, LS
ns
0.422
14, 6
WT and
Trpm5
-/-
, thirst, HS
*
0.034
14, 3
WT and
Trpm5
-/-
, thirst, LS + KCl
***
<0.0001
9, 6
WT and
Trpm5
-/-
, thirst, LS + quinine
***
<0.0001
10, 6
Pdyn
Cre
,
thirst, water
ns
0.9368
5, 4
Pdyn
Cre
, thirst, 0.125 mM quinine
ns
0.9024
6, 5
Pdyn
Cre
, thirst, 0.25 mM quinine
ns
0.8162
5, 4
Ptger3
Cre/wt
, food deprivation, 2 mM AceK
Before LiCl vs After Licl, -Stim
**
0.003
6, 6
Ptger3
Cre/wt
, food deprivation, 2 mM AceK
After Licl, -Stim vs After Licl, +Stim
ns
0.4964
6, 6
Figure 6C
Scramble- vs Ptger3-shRNA
Two-way ANOVA
Two-way ANOVA
Two-way ANOVA
Figure 6E
Scramble-
vs
Ptger3-
shRNA
Ptger3
Cre/
wt
vs
Ptger3
Cre/Cre
Figure 6C
Sidak's multiple comparisons test
Sidak's multiple comparisons test
Figure 7E
Two-way ANOVA
One-way ANOVA (Kruskal–Wallis test)
WT, Dlight
Figure 7G
One-way ANOVA (Kruskal–Wallis test)
Two-way ANOVA
Veh vs PGE2
Veh vs PGE2
Two-way ANOVA
Figure 7A
WT, PGE2 elisa
Figure 7D
Veh vs PGE2
Figure 7F
Veh vs NSAIDs
Two-way ANOVA
Sidak's multiple comparisons test
Sidak's multiple comparisons test
Sidak's multiple comparisons test
Sidak's multiple comparisons test
Dunn’s multiple comparison test
Dunn’s multiple comparison test
Sidak's multiple comparisons test
Dunn’s multiple comparison test
Pdyn
Cre
, HS
One-way ANOVA (Kruskal–Wallis test)
Dunn’s multiple comparison test
Pdyn
Cre
, LS + KCl
One-way ANOVA (Kruskal–Wallis test)
Dunn’s multiple comparison test
Figure S1D
Pdyn
Cre
, LS
One-way ANOVA (Kruskal–Wallis test)
Pdyn
Cre
, LS + quinine
One-way ANOVA (Kruskal–Wallis test)
Figure S3E
-Stim vs +Stim
Two-way ANOVA
Sidak's multiple comparisons test
Figure S3F
+Sodium vs -Sodium
Two-way ANOVA
Sidak's multiple comparisons test
Dunn’s multiple comparison test
Figure S1F
-Stim vs +Stim
Two-way ANOVA
Sidak's multiple comparisons test
Figure S3I
Vch vs CNO
Two-way ANOVA
Sidak's multiple comparisons test
Figure S4A
WT vs
Trpm5
-/-
Two-way ANOVA
Sidak's multiple comparisons test
WT vs
Trpm5
-/-
Two-way ANOVA
Sidak's multiple comparisons test
Figure S4B
-Stim vs +Stim
Two-way ANOVA
Sidak's multiple comparisons test
Figure S4C
One-way ANOVA (Friedman test)
Dunn’s multiple comparison test
Figure S1B
Figure S1C
Thirst vs -Sodium
Thirst vs -Sodium
WT, hot plate
Sated vs -Sodium
Wilcoxon test
NA
ns
0.375
10, 10
Ptger3
Cre/wt
, hot plate, 15 min
-Stim vs +Stim, 15 min
ns
0.3146
4, 4
Ptger3
Cre/wt
, hot plate, 45 min
-Stim vs +Stim, 45 min
ns
0.959
4, 4
WT, von Frey
Sated vs -Sodium
Wilcoxon test
NA
ns
0.25
12, 12
Ptger3
Cre/wt
, von Frey
-Stim vs +Stim
Wilcoxon test
NA
ns
0.625
5, 5
WT, acute pain
ns
>0.9999
4, 4
WT, inflammatory pain
ns
0.2917
4, 4
Ptger3
Cre/wt
, acute pain
ns
0.9665
4, 4
Ptger3
Cre/wt
, inflammatory pain
ns
0.5478
4, 4
LS vs Water
ns
>0.9999
9, 8
LS vs HS
**
0.0013
9, 4
LS vs LS + KCl
*
0.0314
9, 7
LS vs LS + quinine
*
0.0131
9, 9
Trpm5
-/-
, thirst, water
ns
0.2622
6, 5
Trpm5
-/-
, thirst, 500 mM NaCl
ns
0.8475
6, 5
Sated vs -Sodium
ns
0.461
8, 8
Sated vs +Sodium
*
0.0191
8, 7
Sated vs Formain 30 min
ns
0.0804
8, 7
Sated vs Formain 90 min
ns
0.6386
8, 8
Sated vs -Sodium
ns
>0.9999
8, 8
Sated vs +Sodium
ns
>0.9999
8, 8
Sated vs Formain 30 min
ns
0.474
8, 8
Sated vs Formain 90 min
ns
>0.9999
8, 8
Thirst vs Hunger
ns
0.0828
5, 5
Thirst vs Malaise
ns
0.2116
5, 5
Figure S6C
-Sodium, Fos in SFO
Ptger3
Cre/
wt
vs
Ptger3
Cre/Cre
Mann-Whitney test
NA
*
0.0286
4, 4
WT, sated, 500 mM NaCl
ns
0.9973
9, 9
WT, thirst, 500 mM NaCl
***
<0.0001
9, 9
Figure S6F
WT, food deprivation, glucose + quinine
Veh vs PGE2
Wilcoxon test
NA
*
0.0313
6, 6
Figure S6G
Ptger3
Cre/Cre
,
thirst, water
Veh vs PGE2
Wilcoxon test
NA
ns
0.25
4, 4
Figure S6H
WT,
Ptger3-
shRNA, thirst, water
Veh vs PGE2
Mann-Whitney test
NA
ns
0.2108
10, 8
WT, thirst, water
ns
0.9258
5, 5
WT, thirst, 500 mM NaCl
*
0.0455
10, 8
Figure S4D
One-way ANOVA (Friedman test)
Dunn’s multiple comparison test
Figure S4E
Figure S4F
Sated vs Sodium depleted
Two-way ANOVA
Sidak's multiple comparisons test
One-way ANOVA (Kruskal–Wallis test)
Dunn’s multiple comparison test
WT, 5-HT elisa
One-way ANOVA (Kruskal–Wallis test)
Dunn’s multiple comparison test
Figure S4G
-Stim vs +Stim
Two-way ANOVA
Sidak's multiple comparisons test
Figure S5B
Ptger3
Cre/Cre
,
thirst
One-way ANOVA (Kruskal–Wallis test)
Dunn’s multiple comparison test
Figure S6I
Veh vs Formalin
Two-way ANOVA
Sidak's multiple comparisons test
Figure S6B
Ptger3
Cre/wt
, photometry
One-way ANOVA (Kruskal–Wallis test)
Dunn’s multiple comparison test
Figure S6E
Veh vs PGE2
Two-way ANOVA
Sidak's multiple comparisons test
Figure S5C
Scramble-
vs
Ptger3-
shRNA
Two-way ANOVA
Sidak's multiple comparisons test
Figure S6A
WT, progesterone elisa
Table S2. List of genes in Glut1 and Glut2-5 enriched clusters, related to Figure 2.
Enriched genes (alphabetical order)
Glut1
Glut2-5
2900055J20Rik
A730036I17Rik
Nhs
3110035E14Rik
AC124490.1
Nptx1
A830019P07Rik
Ankrd34a
Nrp2
Arhgap36
C130071C03Rik
Ntm
Asic4
Cacnb2
Ntrk3
Bmp3
Calb2
Pard3b
C1ql1
Car2
Pde1a
Cd24a
Cdcp1
Pdzd2
Cdkn3
Chrna7
Penk
D930028M14Rik
Chst2
Rab15
Ddr2
Cntnap5b
Rgs20
Erbb4
Col11a1
Rimbp2
Fam19a1
D930020B18Rik
Rxfp1
Gm30382
Dlgap2
Sema5a
Gm43401
Dpp10
Sertm1
Gpc5
Dusp6
Sh3rf1
Gpd1
Esr1
Slc12a7
Hrk
Fam122b
Slc35f1
Htr7
Fam163a
Slc35f3
Ifit1
Foxp2
Slc35f4
Kazald1
Fzd9
Slc4a4
Kcnk9
Gal
Sorcs1
Krt16
Gm11418
Sorcs2
Lmo2
Gm15631
Spry4
Mapk13
Gm9885
Stk32c
Neurod1
Gpr101
Svil
Ntn1
Gpr149
Thbs2
Onecut2
Gpx3
Tmem132d
Onecut3
Gria1
Tmem163
Pcdh9
Htr2c
Tns3
Pkib
Id4
Trh
Pld5
Il1rap
Vsnl1
Ppp1r17
Il1rapl1
Vstm2a
Ptger3
Il1rapl2
Wipi1
Ptprk
Insm1
Wls
Samd11
Kcnq3
Sncg
Ldb2
Tfap2d
Ltbp1
Tox
Man1c1
Trpc3
Masp1
Wnt4
Mirt1
Mitf
Mpped1
Ncald