nature neuroscience: doi:10.1038/nncpc in the distal, medial and proximal part of dca1. nature...

Supplementary Figure 1

Analyses of dCA1 PC clusters by the Jump method, and principle component analysis (PCA) on different morphological parameters.

(a c) The jump method (see Methods for details) was used to generate the profiles of (a) distortion, (b) transformed distortion, and (c)

jump as a function of the number of cell clusters based on the calculated LRImax and ORImax of apical dendrites from each reconstructed dCA1 PCs. (d,e) 3D scatter plots of dendritic branch points, total length, and branch order of the (d) apical and (e) basal dendrites of all

reconstructed dCA1 PCs, respectively. Each red and blue solid dot represents the morphological data measured from individual cPC

and sPC, respectively. (f h) Principle component analysis of (f) LRImax and ORImax, (g) other 6 morphological parameters including the

total length, total branch points and branch order of the apical and basal dendrite, respectively, and (h) all the above 8 parameters,

respectively, showing the resultant first two principal components. Two groups (black and orange) in each panel are determined by the k-means cluster analysis.

Nature Neuroscience: doi:10.1038/nn.4517


Characterization of intrinsic action potential firing patterns and excitability of dCA1 sPCs and cPCs.

(a1,b1) Representative traces showing action potential firing evoked by a train of ten somatic EPSC-like (tau rise 0.2 ms, tau decay

6 ms) currents (600 2000 pA) of PCs in the (a1) subiculum and (b1) dCA1. The current trains were injected at 5 Hz to the cells.

Asterisks denote the burst spiking. (a2,b2) Summarized results of experiments shown in a1 and b1, indicating (a2) the subiculum regular spiking (black) PCs and bursting PCs (magenta), respectively, (b2) but only regular spiking pattern for both cPCs (red) and

sPCs (blue) in the dCA1. The classification of the regular-spiking and bursting PCs are based on the previous study by Graves et al28

.

Data were presented as mean s.e.m. (c1) Example trace of dCA1 sPC (blue) and cPC (red) action potentials evoked by step current

injection. (c2) Summarized results of firing rate (upper), inter-spike-interval (left bottom) and ratios of the first ISI and the last ISI (testing

spike modulation, right bottom) under increasing step currents. Data were presented as mean s.e.m. P 0.11, measured by Mann-

Whitney U-test.



Differential propagation of dendritic excitation to the soma in sPCs and cPCs revealed by TREES and NEURON simulations.

Simulation was performed under either (a c) MATLAB TREES toolbox or the (d g) NEURON stimulation. (a) A pair of example sPC

and cPC from experimental reconstruction showing dendritic sites of current injection (blue and red dots) and the recording at soma. (b)

Example of somatic voltage change ( V, upper) elicited by injected electronic currents at 450 m apical dendrite (bottom) in the sPC

(blue) and cPC (red), respectively. (c) Cumulative plot of mean V of 171 sPCs (blue) and 183 cPCs (red). Each mean V for a given

PC was averaged from all trials with the injection sites distributed on all distal dendritic sections ( 300 m). Statistic difference was

measured by two-sample Kolmpgorov-Smirnov test, P 0.001. (d) Morphologies of sPC and cPC NERUON models selected from the

ModelDB65,66

depicting distal synaptic inputs, dendritic and somatic recordings. (e,f) Same as b c except using NEURON models. (g)

Comparison of mean somatic and dendritic excitation between sPC and cPC. Data were presented as mean s.e.m. Statistic difference

was measured by unpaired t-test, P 0.001 and P 0.76 for the somatic and local dendritic injections, respectively.



Histological characterization of viral expression of ChR2 in the LEC, MEC and their axon projections along the anterior–posterior axis of CA1.

(a1,a2) Serial sections of LEC (coronal) and MEC (horizontal), demonstrating the ChR2(-mCherry) expression in LEC and MEC (outlined by dashed lines), respectively. Scale bar, 1 mm. (b1) Representative histology verification of ChR2-expressing axons from the

LEC (top) and MEC (bottom) in the hippocampus from anterior (A) to posterior (P) axis, respectively. Scale bar, 1 mm. Note that the ChR2(-mCherry) expressing LEC axons appear an increasing gradient in the CA1 stratum lacunosum-moleculare (SLM) layer along the

A P axis, while that of MEC axons show the opposite pattern. The sections in the dashed line box show the intermingled ChR2-

expressing axons from the LEC and MEC projection neurons. (b2) Higher resolution images showing clearer segregation in the DG MO

layer and distal proximal gradients in the CA1 SLM layer of LEC and MEC axons in the boxed regions in b1, Scale bar, 200 m.



Characterization of optogenetic activation of EC–dCA1 monosynaptic transmission by AOD-based laser stimulation system.

(a) Representative traces of EPSCCRACM recorded from the CA1 PC (blue) and the CA2 GABAergic interneuron (red), which,

respectively, show consistent mono-synaptic responses and variable poly-synaptic responses, evoked by 473-nm laser pulses with

increasing intensities in the CRACM experiment. (b,c) Effects of applying GABAARs blocker picrotoxin (50 M) on (b) the responsive

map, (c) EPSCCRACM amplitudes and the response latency in the dCA1 PC. P 0.29 and P 0.004 for the amplitude and latency,

respectively, calculated by the paired t-test. (d,e) Effects of application of TTX (0.5 M) and 4-AP (100 M) on (d) the discrete dendritic

EPSCCRACM map, EPSCCRACM latency (bottom: averaged EPSCCRACM traces from the indicated locations in the map), and (e) the mean

maximal amplitude of EPSCCRACM. Data were presented as mean s.e.m. P 0.01, measured by paired t-test. (f,g) Effects of HCN

channel blocker ZD7288 (20 M), slimilar as d,e. Note a slight increase of mean amplitude of EPSCCRACM after the ZD compound

application. Data were presented as mean s.e.m. P 0.16, calculated by the paired t-test. (h) Blocking the GABAARs with picrotoxin

(50 M) does not affect the differential innervations of direct excitatory TA inputs from LEC to dCA1, compared to the data in Fig. 2e. P

3.8 10-8

, measured by Person’s Chi-square test. (i) Preserved differential innervations of LEC and MEC direct inputs to sPC and

cPC in the distal, medial and proximal part of dCA1.



Direct long-range GABAergic transmission from LEC and MEC to dCA1 pyramidal neurons.

(a) Example IPSCCRACM and responsive map of direct GABAergic inputs from the MEC to a dCA1 PC, measured under 0 mV

membrane potential and presence of CNQX (10 M) and AP5 (50 M). (b) Percentage of cell receiving direct GABAergic projections

from LEC or MEC in the dCA1 sPCs and cPCs as well as dCA2 PCs. n, number of tested cells.



Comparison of transmission strength of CA3 SC excitatory synapses on dCA1 sPCs and cPCs.

(a) Diagram of recording configuration, in which the neighboring sPC and cPC were simultaneously recorded and a filed stimulation

electrode (concentric tungsten electrode) was placed in a distance of 350 m away in the middle of SR layer. (b) Example EPSC

traces recorded from sPC and cPC under increasing stimulus intensities. (c) Summarized EPSC amplitude ratios of the cPC and sPC

from the assay shown in a and b. Data were presented as mean s.e.m.



Characterization of Calb2-IRES-Cre::Ai9 and Calb1-2A-dgCre::Ai9 mice.

(a,b) Example images showing the genetically-tdTomato labeled cells (red) immunostained with monoclonal antibody against calbindin-D28k (green) in the dCA1 of (a) Calb2-IRES-Cre::Ai9 and (b) Calb1-2A-dgCre::Ai9 (7-d TMP induction, see Methods) mice. Scale bar,

100 m. (c,d) Characterization of hippocampal labeling from anterior (A) to posterior (P) in (c) Calb2-IRES-Cre::Ai9 and (d) Calb1-2A-

dgCre::Ai9 (7-d induction) mice. Scale bar, 1 mm. All following statistic results are measured in the sections from Bregma -2.0 to -3.5

mm with 0.3 mm step along the A P axis. (e,f) Comparison of the labeling efficiency (yellow/green) and colocalization rate (yellow/red)

in the (e) CA1 SP and (f) DG between the Calb2-IRES-Cre::Ai9 and Calb1-2A-dgCre::Ai9 (with 1 or 7 d induction) mice, respectively. (g,h) Fluorescence images showing the (g) neuronal GABA staining in the dCA1 of Calb2-IRES-Cre::Ai9, and the (h) tdTomato labeling

rates in GABAergic cells in different layers of dCA1. Note the co-localization rates are 10 . Scale bar, 100 m. (i,j) Similar as g,h

except for Cre antibody staining. Note that Calb2-IRES-Cre::Ai9 mice exhibit high efficiency ( 90 ) in labeling neurons in the dCA1 SP

layer, but low efficiency ( 15 ) in labeling neurons in other layers. Scale bar, 100 m. Data were presented as mean s.e.m.



HSV-1-G3 anterograde transportation from dorsal or ventral LEC and MEC to hippocampal CA1.

(a,b) Co-localization of anterograde transportation of HSV-1-G3 (green) from (a) anterior ventral or posterior dorsal LEC; (b) ventral or

dorsal MEC to hippocampal CA1 in Calb2-IRES-Cre::Ai9 transgenic mice (tdTomato). Scale bar, 100 m. (c) Summarized co-localized

rates following the HSV-1-G3 injected to different parts of MEC or LEC, respectively. Note that no significant difference was observed

between the dorsal and ventral parts within the LEC or MEC. Data were presented as mean s.e.m. (d,e) Characterization of HSV

injected in the (d) anterior ventral or posterior dorsal LEC and (e) ventral or dorsal MEC in Calb2-IRES-Cre::Ai9 transgenic mice. Scales bar, 1 mm. (f) Serial brain sections showing the spreading infection area of HSV-1-G3 3 days after the injection in the anterior-

ventral LEC. Scale bar, 1 mm.



Postbehavior histological characterization of NpHR-EYFP expression in LEC or MEC and the locations of implanted optical fibers or optetrodes in dCA1.

(a) Post-histology of the mice with AAV-CaMKII -NpHR-EYFP or AAV-CaMKII -EYFP injected in the bilateral LEC and subsequently

behaviorally tested shown in Fig. 5c. The first column: atlas of different coronal sections; 2nd column: example fluorescence images of

NpHR-EYFP expression in the bilateral LEC and optical fiber implanted in the dCA1 (scale bar, 1 mm); 3rd column: overlaid image showing the region of NpHR-EYFP expression of all tested mice; 4th column, overlaid image showing the region of EYFP expression of all tested control mice. Grey gradients: viral expression levels in all tested mice, red gradients: optical fiber tracks. (b) Similar as a except for the optical fiber implantation was made in the DG in mice, whose results are shown in Fig. 5d. (c) Similar as a except for the virus was injected in the MEC, whose results are shown in Fig. 5e. (d) Similar as a except for the mice whose results are shown in Fig.

5f h, respectively. (e) Fluorescence images of hippocampal sections show the expression of Arch-GFP (green) and the optetrode

recording sites (arrows) in the dCA1. Uppers 4 mice were recorded in the right hemisphere, while the bottoms 6 were done in the left hemisphere. Scale bar, 1 mm.



Similar impairments of reversed learning by inactivation of LEC–dCA1 transmissions or of postsynaptic dCA1 Calb+ cPCs.

(a1 3) The inactivation of (a1) LEC dCA1, but not (a2) LEC DG or (a3) MEC dCA1 transmissions impairs the reverse learning, in

which the odor licking/non-licking contingencies were reversed, in the days 4 6 with the same mice which underwent the initial learning

shown in Fig. 5c e, respectively. Data were presented as mean s.e.m. (a4 5) Suppressing Calb+ PCs activities in the (a4) dCA1, but

not in the (a5) DG impairs the reversal learning in the days 4 6 in the same mice which underwent the initial learning shown in Fig. 5f

and g, respectively. Data were presented as mean s.e.m. (b1 5) Detailed changes of the hit rate (upper) and the correct rejection rate

(bottom) for behavioral performance data shown in the Fig. 5c g and Supplementary Fig. 12a1 5 during the initial and reversal

learning, respectively. Statistic differences were presented in the Supplementary Table 4 15. Data were presented as mean s.e.m.



Optogenetic tagging of dCA1 Calb+ cPC units with Arch in vivo in comparison with that of tagging with ChR2.

(a) Comparisons of waveforms of the spontaneous spikes (top traces) and the optical ChR2-excitation induced spikes (by 5 ms blue

laser pulses, 0.5 mW, bottom) recorded from dCA1 PV+IN and PCs with the optetrode in the Pavlb-Cre::Ai27 and Calb2-IRES-Cre::Ai27

mice, respectively. Note that the optical ChR2-excitation significantly alters the waveform of PCs’ spikes, but not that of PV+INs,

indicated by cross correlation (cc) values (r). (b,c) Example recording of the LFPs and spike units from (b) PCs and (c) PV+INs with 5

ms blue-laser pulse stimulation (0.5 mW) in the dCA1 area of the Calb2-IRES-Cre::Ai27 and Pavlb-Cre::Ai27 mice, respectively.

Waveforms: averaged spontaneous spikes (left), individual ChR2-induced spikes (blue) or spontaneous spikes (black, corresponding to

the *-labeled spikes in the middle). r: cc between the averaged spike and the individual spike. (d) Similar to b c, but showing the

example of two tagged dCA1 Calb+

PCs with Arch in Calb2-IRES-Cre::Ai35 mice. Note that the yellow-laser illumination (2 min, 10 mW, indicated by yellow bars) significant suppresses the rate of spontaneous spikes, but exhibiting identical waveforms recorded during the

ON and OFF of laser stimulation. (e) Example of simultaneous recordings from the Calb+Arch

+ cPC (red) and Calb Arch sPC (blue) in

hippocampal slices of the Calb2-IRES-Cre::Ai35 mouse, showing differential effects of 2-min yellow-light illumination (10 mW, indicated by the yellow bar) on evoked rhythmic spikes (by intracellular injection of 5 ms current pulses at 0.5 Hz, black traces) between these two subtypes of dCA1 PCs. (f) Statistic results of the changes of spike rate and resting membrane potential during the ON and OFF of

the yellow-light illumination in the Calb+Arch

+ cPC (red) and Calb Arch sPC (blue) in the experiments shown in d. P 0.008, measured

by the Wilcoxon signed-ranked test. (g) Example recordings and (h) statistical results showing changes of the frequency and amplitude

of spontaneous IPSCs (Vclamp 0 mV) during the ON and OFF of 2-min laser illumination in the dCA1 Calb+Arch

+ cPC (red colors) and

Calb Arch sPC (blue colors) in the slices of Calb2-IRES-Cre::Ai35 mice.



Varied spiking responses and selectivities in response to odors A and B during odor sampling in trials.

(a) Raster plots of spikes (upper two panels) and peri-stimuli time plots of firing rate changes (in 100 ms bins) for single-units recorded

in the Go (red, odor A) and No-go (blue, odor B) trials in different mice (see the unit identity). The odor delivery duration is indicated by the green bar. The numbers for individual types are presented in the Supplementary Table 16.



Comparison of neuronal spiking activities in Correct and Error trials in the behavioral task.

(a) Comparison of the mean firing rates of all PC units (left), sPC (middle), or cPC (right) in the Correct and Error trials in Go (upper) and No-go trials (bottom), respectively. The upper dots indicate the significant differences between Correct and Error trials (Paired t-

test, P 0.05). Data were presented as mean s.e.m. (b) D’-prime analysis of differential firing of individual units in Correct and Error

trials in Go (left) and No-go trials (right), respectively. Red and blue columns in the upper right indicate the cPC and sPC, respectively and sorted by the mean d’-prime of 1.5-s after odor delivery. The lower panels are the mean population d’-prime of sPC (blue) and cPC (red) during Correct and Error trials in Go (left) and No-go trials (right), respectively. Black dashed line, 95% chance level. All units were recorded in the first learning day.



Comparison of spontaneous firing rates during different brain states between the identified Calb+ and Calb dCA1 PCs.

(a) Examples of neuronal spikes of units and local field potentials in the running, rapid-eye movement (REM) sleep and slow-wave

sleep (SWS) states, respectively. Asterisks label the units of light-responsive Calb+ cells. Scale bar, 1 sec (middle panel) or 100 ms

(enlarged, bottom panel), 0.5 mV. (b) Distributions of Calb+ (red) and Calb (blue) dCA1 PCs showing different spontaneous firing rates

during the RUN, REM, SWS, respectively. P 0.0127, 0.0282 and 0.0025 for RUN, SWS and REM state, respectively, calculated by

the Mann-Whitney U-test. Note that this result is consistent with a previous study by Mizuseki et al29

.


Supplementary Table 1 Comparisons of dendritic arborizations between the cPC and sPC in

the dCA1.

Dendritic morphology parameters cPC (n = 183) sPC (n = 171) Statistical difference

Total apical dendrite length (µm) 3353 ± 58 2576 ± 48 P = 1.84 × 10-24

Apical branch order 16.40 ± 0.25 18.40 ± 0.34 P = 3.17 × 10-6

Apical branch points 35.79 ± 0.74 27.45 ± 0.64 P = 9 × 10-20

Total basal dendrite length (µm) 1868 ± 50 1730 ± 46 P = 0.042

Basal dendrite branch order 6.10 ± 0.14 6.10 ± 0.11 n.s.

Basal dendrite branch points 16.0 ± 0.45 16.9 ± 0.52 n.s.

Unpaired t-test is used to measure significance of the difference in the cell numbers between groups. n.s.: no

significance.

Supplementary Table 2 Statistical comparison of cell clustering using the principal component

analysis (PCA) with different dendritic morphology parameters.

Data source for PCA and cluster analysis

Distance between 2 centers

Mean distance to centers

# of cells in each cluster

% of variance explained by

first 2 PCs Cluster 1 Cluster 2 Cluster 1 Cluster 2 LRImax and ORImax 1.390 0.093 0.146 184 170 100% 6 morphological

parameters 1.091 0.241 0.335 203 151 84.62%

LRImax, ORImax and 6 morphological

parameters 1.502 0.515 0.449 186 168 81.62%

It refers to the data shown in Supplementary Fig. 1f-h. The other 6 dendritic morphological parameters are the total

apical length, the total apical branch points, the maximum apical branch order, the total basal length, the total basal

branch points, and the maximum basal branch order.


Supplementary Table 3 Comparisons of basic membrane and spiking properties among cPCs

and sPCs in the dCA1, CA2 PCs and subiculum PCs.

CA1 CA2 Subiculum

cPC (n = 70) sPC (n = 84) PC (n = 51) Regular (n = 14) Burst (n = 15)

Cm (pF) 113.80 ± 3.40 97.45 ± 3.20*** 135.75 ± 5.89 129.26 ± 6.98 137.10 ± 6.98

Rm (MΩ) 72.35 ± 2.31 85.32 ± 3.35** 51.94 ± 2.17 64.66 ± 6.41 50.24 ±4.97

Tau (ms) 1.72 ± 0.07 1.51 ± 0.07 1.64 ± 0.10 2.21 ± 0.07 2.10 ± 0.09 Spike threshold

(mV) 44.22 ± 0.35 43.32 ± 0.36 40.51 ± 0.70 40.89 ± 0.91 44.87 ± 0.50

Spike amplitude (mV) 99.78 ± 0.74 99.13 ± 0.61 88.87 ± 1.42 87.50 ± 2.94 88.87 ± 2.48

Spike peak (mV) 55.05 ± 0.72 55.81 ± 0.58 48.36 ± 0.93 46.61 ± 2.50 40.00 ± 2.54 Spike half-width

(ms) 1.31 ± 0.04 1.30 ± 0.05 1.12 ± 0.05 1.57 ± 0.06 1.30 ± 0.06

Max dv/dt (mV/ms) 314.35 ± 6.83 317.46 ± 7.80 272.80 ± 9.11 204.99 ± 18.76 209.48 ± 18.63

Min dv/dt (mV/ms) 68.79 ± 2.15 68.79 ± 2.16 65.92 ± 2.21 49.26 ± 2.28 26.14 ± 5.08

Spike area (mV*ms) 125.19 ± 4.32 121.68 ± 4.39 97.00 ± 3.97 128.99 ± 4.03 107.62 ± 3.31

After-hyperpolarization

(mV) 8.50 ± 0.28 9.03 ± 0.22 9.08 ± 0.32 9.25 ± 0.45 8.41 ± 0.08

Max Ih (pA) 253.51 ± 13.30 244.41 ± 11.26 164.56 ± 12.57

I/O slop (Hz/pA) 0.0482 ± 0.0013 0.0497 ± 0.0013 0.0164 ± 0.0016 Significant difference between cPCs and sPCs was only observed in membrane capacitance (Cm) and resistance

(Rm), ***: P < 0.001, **: P = 0.0026; unpaired t-test. Most parameters of the dCA1 cPCs and sPCs are significantly

different from that of the CA2 and subiculum PCs (P values are not shown), which are classified into two sub-

populations: regular- and burst-spiking PCs, according to the paper by Graves et al28.


Supplementary Table 4 Statistical results on the behavioral performance, hit and

correct rejection rates in NpHR and EYFP (control) groups in the first 2-days of

learning.

Data source Statistic method

Statistic results Mice number

Performance correct rate NpHR vs EYFP LEC injection CA1 implant

Two-way mixed design ANOVA

Ftype (1,18) = 72.82; Ptype = 9.7×10-8 Fblock (19,342) = 23.57; Pblock = 5.8×10-51 Finteraction(19,342) = 6.40; Pinteraction = 3.5×10-14

NpHR=10; EYFP=10

Hit rate NpHR vs EYFP LEC injection CA1 implant


Ftype (1,18) = 15.15; Ptype = 0.001 Fblock (19,342) = 7.03; Pblock = 7.9×10-16 Finteraction(19,342) = 2.03; Pinteraction = 0.007;

NpHR=10; EYFP=10

Correct rejection rate NpHR vs EYFP LEC injection CA1 implant


Ftype (1,18) = 13.40; Ptype = 0.002 Fblock (19,342) = 34.20; Pblock = 2.4×10-67 Finteraction(19,342) = 2.63; Pinteraction = 2.8×10-4

NpHR=10; EYFP=10

Performance correct rate NpHR vs EYFP LEC injection DG implant


Ftype (1,17) = 0.64; Ptype = 0.435 Fblock (19,323) = 75.65; Pblock= 1.3×10-106 Finteraction(19,323) = 0.74; Pinteraction = 0.78

NpHR=10; EYFP=9

Hit rate, NpHR vs EYFP LEC injection DG implant


Ftype (1,17) = 2.97; Ptype = 0.103 Fblock (19,323) = 5.58; Pblock = 6.2×10-12 Finteraction(19,323) = 2.02; Pinteraction = 0.007

NpHR=10; EYFP=9

Correct rejection rate, NpHR vs EYFP DG implant


Ftype (1,17) = 1.39; Ptype = 0.255; Fblock (19,323) = 55.30; Pblock = 1.9×10-89 Finteraction(19,323) = 1.05; Pinteraction = 0.398

NpHR=10; EYFP=9

Performance correct rate NpHR vs EYFP MEC injection CA1 implant


Ftype (1,18) = 0.001 ; Ptype = 0.975 Fblock (19,342) = 62.69 ; Pblock = 4.7×10-99 Finteraction(19,342) = 0.77 ; Pinteraction =0.749

NpHR=10; EYFP=10

Hit rate NpHR vs EYFP MEC injection CA1 implant


Ftype (1,18) = 0.86 ; Ptype = 0.365 Fblock (19,342) = 6.96; Pblock = 1.18×10-15 Finteraction(19,342) = 0.80; Pinteraction = 0.702 ;

NpHR=10; EYFP=10

Correct rejection rate NpHR vs EYFP MEC injection CA1 implant


Ftype (1,18) = 0.22 ; Ptype = 0.647; Fblock (19,342) = 35.02 ; Pblock = 1.80×10-68 Finteraction(19,342) = 0.93; Pinteraction = 0.785

NpHR=10; EYFP=10

It refers to the data shown in (Fig. 5c-e and Supplementary Fig. 12b1-3). NpHR group: LEC or MEC

injection with AAV-CaMKIIα-eNpHR-EYFP; EYFP control group: LEC or MEC injection with AAV-

CaMKIIα-EYFP. Optical fibers were implanted in either the dCA1 or the DG. All analyses were based on blind

experimental design.


Supplementary Table 5 Statistical results on the behavioral performance, hit and

correct rejection rates in Arch+ and Ai35 (control) groups in the first 2-days of

learning.

Data source Statistic method Statistic results Mice number

Performance correct rate Arch+ vs Ai35, CA1 implant


Ftype(1,18) = 10.45; Ptype = 0.005 Fblock(19,342) = 47.61; Pblock = 6.0×10-84 Finteraction(19,342)=1.80; Pinteraction=0.022

Arch+=10 Ai35=10

Hit rate Arch+ vs Ai35, CA1 implant


Ftype(1,18) = 7.14 ; Ptype = 0.016 Fblock(19,342) = 5.18; Pblock = 5.9×10-11 Finteraction(19,342) = 1.30 ; Pinteraction = 0.178

Arch+=10 Ai35=10

Correct rejection rate Arch+ vs Ai35 CA1 implant


Ftype(1,18) = 1.85; Ptype = 0.191 Fblock(19,342) = 28.20; Pblock = 1.6×10-58 Finteraction(19,342) =1.45; Pinteraction = 0.101

Arch+=10 Ai35=10

Performance correct rate Arch+ vs Ai35 DG implant


Ftype(1,19) = 2.19; Ptype = 0.155 Fblock(19,361) = 64.82; Pblock = 8.1×10-104 Finteraction(19,361) = 1.11; Pinteraction = 0.342

Arch+=11 Ai35=10

Hit rate Arch+ vs Ai35 DG implant


Ftype(1,19) = 0.33; Ptype = 0.574 Fblock(19,361) = 6.34 ; Pblock = 3.8×10-14 Finteraction(19,361) = 0.53 ; Pinteraction = 0.950

Arch+=11 Ai35=10

Correct rejection rate Arch+ vs Ai35 DG implant


Ftype(1,19) = 3.10; Ptype = 0.095 Fblock(19,361) = 46.88 ; Pblock = 3.0×10-85 Finteraction(19,361) = 1.08; Pinteraction = 0.370

Arch+=11 Ai35=10

It refers to the data shown in (Fig. 5f-g and Supplementary Fig. 12b4-5). Arch+ group: Calb2-IRES-

Cre::Ai35 mice; Control: Ai35 mice. Optical fibers were implanted in either the dCA1 or the DG. All

analyses were based on blind experimental design.


Supplementary Table 6 Statistical results on the behavioral performance in NpHR

and EYFP (control) mouse groups in the 3rd days of learning.

Data source Statistic method Statistic results Mice number Performance correct rate laser on vs laser off, NpHR mice LEC injection, CA1 implant

Mann-Whitney U-test (use block averaged data)

P = 0.036

NpHR=10

Performance correct rate laser on vs laser off, EYFP mice LEC injection, CA1 implant

Mann-Whitney U-test

P =0.023 (performance increase)

EYFP=10

Performance correct rate NpHR mice laser on vs EYFP mice laser on LEC injection, CA1 implant

Mann-Whitney U-test

P = 1.678×10-4

NpHR=10 EYFP=10

Performance correct rate NpHR mice laser off vs EYFP mice laser off, LEC injection, CA1 implant

Mann-Whitney U-test

P = 0.033 (performance increase)

NpHR=10 EYFP=10

Performance correct rate laser on vs laser off, NpHR mice LEC injection, DG implant

Mann-Whitney U-test

P = 0.048 (performance increase in ON)

NpHR=10

Performance correct rate laser on vs laser off, EYFP mice LEC injection, DG implant

Mann-Whitney U-test

P = 0.211

EYFP=9

Performance correct rate NpHR mice laser on vs EYFP mice laser on, LEC injection, DG implant

Mann-Whitney U-test

P = 0.429;

NpHR=10 EYFP=9

Performance correct rate NpHR mice laser off vs EYFP mice laser off, LEC injection, DG implant

Mann-Whitney U-test

P = 0.787

NpHR=10 EYFP=9

Performance correct rate laser on vs laser off, NpHR mice MEC injection, CA1 implant

Mann-Whitney U-test

P = 0.002; (performance increase in ON)

NpHR=10

Performance correct rate laser on vs laser off, EYFP mice MEC injection, CA1 implant

Mann-Whitney U-test

P = 0.008 (performance increase in ON)

EYFP=10

Performance correct rate NpHR mice laser on vs EYFP mice laser on, MEC injection, CA1 implant

Mann-Whitney U-test

P = 0.878 ;

NpHR=10 EYFP=10

Performance correct rate NpHR mice laser off vs EYFP mice laser off, MEC injection, CA1 implant

Mann-Whitney U-test

P = 0.703;

NpHR=10 EYFP=10

It refers to the data shown in (Fig. 5c-e). NpHR group: LEC or MEC injection with AAV-CaMKIIα-eNpHR-

EYFP; EYFP control group: LEC or MEC injection with AAV-CaMKIIα-EYFP. Optical fibers were implanted

in either the dCA1 or the DG. All analyses were based on blind experimental design.


Supplementary Table 7 Statistical results on hit rate and correct rejection (CR)

rate in NpHR and EYFP groups in the 3rd day of learning.

It refers to the data shown in (Supplementary Fig. 12b1-3).

Data source Statistic method Statistic results Mice number Hit and CR rates laser on vs laser off, NpHR mice LEC injection, CA1 implant


P Hit = 0.246 P CR = 0.056

NpHR=10

Hit and CR rates laser on vs laser off, EYFP mice LEC injection, CA1 implant

Mann-Whitney U-test

P Hit = 0.278 P CR = 0.039 (performance increase)

EYFP=10

Hit and CR rates NpHR mice laser on vs EYFP mice laser on LEC injection, CA1 implant

Mann-Whitney U-test

P Hit = 0.007 P CR = 1.62×10-

4

NpHR=10; EYFP=10

Hit and CR rates NpHR mice laser off vs EYFP mice laser off LEC injection, CA1 implant

Mann-Whitney U-test

P Hit = 0.435 P CR = 0.268

NpHR=10; EYFP=10

Hit and CR rates laser on vs laser off, NpHR mice LEC injection, DG implant

Mann-Whitney U-test

P Hit = 0. 593 P CR = 0.234

NpHR=10;

Hit and CR rates laser on vs laser off, EYFP mice LEC injection, DG implant

Mann-Whitney U-test

P Hit = 0.471 P CR = 0.364

EYFP=9;

Hit and CR rates NpHR mice laser on vs EYFP mice laser on LEC injection, DG implant

Mann-Whitney U-test

P Hit = 0.248 P CR = 0.213

NpHR=10; EYFP=9;

Hit and CR rates NpHR mice laser off vs EYFP mice laser off LEC injection, DG implant

Mann-Whitney U-test

P Hit = 0.517 P CR = 0.649

NpHR=10; EYFP=9;

Hit rate and CR rates laser on vs laser off, NpHR mice MEC injection, CA1 implant


P Hit = 0.265; P CR = 0.031

NpHR=10

Hit rate and CR rates laser on vs laser off, EYFP mice MEC injection, CA1 implant

Mann-Whitney U-test


EYFP=10

Hit rate and CR rates NpHR mice laser on vs EYFP mice laser on, MEC injection, CA1 implant

Mann-Whitney U-test

P Hit = 0.925; P CR= 0.760

NpHR=10 EYFP=10

Hit rate and CR rates NpHR mice laser off vs EYFP mice laser off, MEC injection, CA1 implant

Mann-Whitney U-test

P Hit = 0.533; P CR = 0.970

NpHR=10 EYFP=10


Supplementary Table 8 Statistical results on the behavioral performance in Arch+ and

Ai35 (control) groups in the 3rd day of learning.


Performance correct rate laser on vs laser off Arch+ mice CA1 implant


P =0.031

Arch+ =10

Performance correct rate laser on vs laser off Ai35 mice CA1 implant

Mann-Whitney U-test

P =0.180

Ai35 =10

Performance correct rate Arch+ mice laser on vs Ai35 mice laser on, CA1 implant

Mann-Whitney U-test

P =0.036

Arch+ =10 Ai35 =10

Performance correct rate Arch+ mice laser off vs Ai35 mice laser off, CA1 implant

Mann-Whitney U-test

P =0.701

Arch+ =10 Ai35 =10

Performance correct rate laser on vs laser off Arch+ mice DG implant

Mann-Whitney U-test


Arch+ =11

Performance correct rate laser on vs laser off Ai35 mice DG implant

Mann-Whitney U-test


Ai35 =10

Performance correct rate Arch+ mice laser on vs Ai35 mice laser on DG implant

Mann-Whitney U-test

P =0.617

Arch+ =11 Ai35 =10

Performance correct rate Arch+ mice laser off vs Ai35 mice laser off DG implant

Mann-Whitney U-test

P =0.915

Arch+ =11 Ai35 =10

It refers to the data shown in (Fig. 5f-g).


Supplementary Table 9 Statistical results on hit rate and correct rejection (CR) rate

in Arch+ and Ai35 groups in the 3rd day of learning.


Hit and CR rates laser on vs laser off Arch+ mice CA1 implant


P Hit = 0.551 P CR = 0.036

Arch+ =10

Hit and CR rates laser on vs laser off Ai35 mice CA1 implant

Mann-Whitney U-test

P Hit = 0.077 P CR = 0.220

Ai35 =10

Hit and CR rates Arch+ mice laser on vs Ai35 mice laser on CA1 implant

Mann-Whitney U-test

P Hit = 0.078 P CR = 0.018

Arch+ =10 Ai35 =10

Hit and CR rates Arch+ mice laser off vs Ai35 mice laser off CA1 implant

Mann-Whitney U-test

P Hit = 0.764 P CR = 0.113

Arch+ =10 Ai35 =10

Hit and CR rates laser on vs laser off Arch+ mice DG implant

Mann-Whitney U-test

P Hit = 0.099 P CR = 0.111

Arch+ =11

Hit and CR rates laser on vs laser off Ai35 mice DG implant

Mann-Whitney U-test

P Hit= 0.834 P CR = 0.009 (performance increase)

Ai35 =10

Hit and CR rates Arch+ mice laser on vs Ai35 mice laser on DG implant

Mann-Whitney U-test

P Hit = 0.152; P CR = 0.240;

Arch+ =11 Ai35 =10

Hit and CR rates Arch+ mice laser off vs Ai35 mice laser off DG implant

Mann-Whitney U-test

P Hit = 0.616; P CR = 0.915;

Arch+ =11 Ai35 =10



Supplementary Table 10 Statistical results on the behavioral performance, hit and correct

rejection rates in NpHR and EYFP (control) mouse groups in the (reserved) learning days

4-5.



Performance correct rate NpHR vs EYFP LEC injection CA1 implant


Ftype (1,17) = 75.1; P type = 1.2×10-7 Fblock (19,323) = 100.5; P block = 3.6×10-89 Finteraction(19,323) = 2.1; P interaction = 0.005

NpHR=10 EYFP=10

Hit rate NpHR vs YFP LEC injection CA1 implant


FeNpHR3.0(1,17) = 11.9; P eNpHR = 0.003 Fblock (19,323) = 69.2; P block = 1.3×10-101 Finteraction(19,323) = 1.7; P interaction = 0.04

NpHR=10 EYFP=10

Correct rejection rate in acquisition, NpHR vs YFP LEC injection CA1 implant


FeNpHR(1,17) = 8.8; P eNpHR = 0.009 Fblock (19,323) = 23.3; P block = 1.6×10-49 Finteraction(19,323) = 3.5; P interaction = 2.5×10-6

NpHR=10 EYFP=10

Performance correct rate in acquisition, reverse NpHR vs EYFP LEC injection DG implant


Ftype (1,16) = 1.8 × 10-4; P type = 0.99 Fblock (19,304) = 141.5; P block = 1.7×10-138 Finteraction(19,304) = 1.02; P interaction = 0.43

NpHR=10 EYFP=9

Hit rate in acquisition, NpHR vs YFP LEC injection DG implant


FeNpHR3.0(1,16) = 3.7; P eNpHR = 0.07 Fblock (19,304) = 90.8; P block = 5.0×10-113 Finteraction(19,304) = 1.4; P interaction = 0.11

NpHR=10 EYFP=9

Correct rejection rate in acquisition, NpHR vs YFP LEC injection DG implant


FeNpHR3.0(1,16) = 0.92; P eNpHR3.0 = 0.35 Fblock (19,304) = 29.2; P block = 2.6×10-57 Finteraction(19,304) = 0.63; P interaction = 0.89

NpHR=10 EYFP=9

Performance correct rate NpHR vs EYFP MEC injection CA1 implant


Ftype (1,18) = 0.348; Ptype = 0.562 Fblock (19,342) = 87.543 ; Pblock = 9.204×10-

119 Finteraction(19,342) = 0.656; Pinteraction = 0.861

NpHR=10; EYFP=10

Hit rate NpHR vs EYFP MEC injection CA1 implant


Ftype (1,18) = 0.263; Ptype = 0.614; Fblock (19,342) = 45.045 ; Pblock = 4.827×10-81 Finteraction(19,342) = 0.465; Pinteraction = 0.974;

NpHR=10; EYFP=10

Correct rejection rate NpHR vs EYFP MEC injection CA1 implant


Ftype (1,18) = 1.045×10-4; Ptype = 0.992; Fblock (19,342) = 22.031; Pblock =2.880×10-48 ; Finteraction(19,342) = 0.517; Pinteraction = 0.955;

NpHR=10; EYFP=10

It refers to the data shown in (Supplementary Fig. 12a1-3 and b1-3).


Supplementary Table 11 Statistical results on behavioral performance, hit and

correct rejection rates in Arch+ and Ai35 groups in the (reversed) learning days 4-5.



Performance correct rate Arch+ vs Ai35 CA1 implant


Ftype(1,18) = 23.8; P type = 1.2×10-

4 Fblock(19,342) = 81.4; P block = 2.4×10-114 Finteraction(19,342) = 5.3; Pinteraction = 4.5×10-

11

Arch+=9 Ai35 =9

Hit rate Arch+ vs Ai35 CA1 implant


Ftype(1,18) = 13.1; P type = 0.002 Fblock(19,342) = 124.3; P block = 8.9×10-141 Finteraction(19,342) = 2.9; P interaction = 5.4×10-5

Arch+=9 Ai35 =9

Correct rejection rate Arch+ vs Ai35 CA1 implant


Ftype(1,18) = 7.3; P type = 0.015 Fblock(19,342) = 16.4; P block = 1.2×10-37 Finteraction(19,342) = 3.7; P interaction = 5.0×10-7

Arch+=9 Ai35 =9

Performance correct rate Arch+ vs Ai35, DG implant


Ftype(1,19) = 0.95; P type = 0.34 Fblock(19,361) = 135.6; P block = 2.4×10-151 Finteraction(19,361) = 0.85; P interaction = 0.65

Arch+=11 Ai35 =10

Hit rate Arch+ vs Ai35 DG implant


F type(1,19) = 2.206; P type = 0.15 Fblock(19,361) = 112.2; P block = 1.5×10-138 Finteraction(19,361) = 2.1; P interaction = 0.005

Arch+=11 Ai35 =10

Correct rejection rate Arch+ vs Ai35, DG implant


F type(1,19) = 2.97; P type = 0.10 Fblock(19,361) = 30.2; P block = 7.2×10-63 Finteraction(19,361) = 1.07; P interaction = 0.38

Arch+=11 Ai35 =10

It refers to the data shown in (Supplementary Fig. 12a4-5 and b4-5).


Supplementary Table 12 Statistical results on the behavioral performance in NpHR and

EYFP (control) mouse groups in the (reserved) learning day 6.


Performance correct rate laser on vs laser off, NpHR mice LEC injection, CA1 implant


P = 0.015

NpHR =10

Performance correct rate laser on vs laser off, EYFP mice LEC injection, CA1 implant

Mann-Whitney U-test

P = 0.388

EYFP =9

Performance correct rate NpHR mice laser on vs EYFP mice laser on LEC injection, CA1 implant

Mann-Whitney U-test

P = 6.5×10-4

NpHR =10 EYFP =9

Performance correct rate NpHR mice laser off vs EYFP mice laser off LEC injection, CA1 implant

Mann-Whitney U-test

P = 0.042

NpHR =10 EYFP =9

Performance correct rate laser on vs laser off, NpHR mice LEC injection, DG implant

Mann-Whitney U-test

P = 0.028 (increased performance)

NpHR =9

Performance correct rate laser on vs laser off, EYFP mice LEC injection, DG implant

Mann-Whitney U-test

P = 0.117

EYFP =10

Performance correct rate NpHR mice laser on vs EYFP mice laser on LEC injection, DG implant

Mann-Whitney U-test

P = 0.681

NpHR =9 EYFP =10

Performance correct rate NpHR mice laser off vs EYFP mice laser off LEC injection, DG implant

Mann-Whitney U-test

P = 0.837

NpHR =9 EYFP =10

Performance correct rate laser on vs laser off, NpHR mice MEC injection, CA1 implant


P = 0.015 ; (increased performance)

NpHR=10

Performance correct rate laser on vs laser off, EYFP mice MEC injection, CA1 implant

Mann-Whitney U-test

P = 0.136 ;

EYFP=10

Performance correct rate NpHR mice laser on vs EYFP mice laser on, MEC injection, CA1 implant

Mann-Whitney U-test

P = 0.169 ;

NpHR=10 EYFP=10

Performance correct rate NpHR mice laser off vs EYFP mice laser off, MEC injection, CA1 implant

Mann-Whitney U-test

P = 0.939 ;

NpHR=10 EYFP=10

It refers to the data shown in (Supplementary Fig. 12a1-3).


Supplementary Table 13 Statistical results on hit rate and correct rejection (CR) rates in

NpHR and EYFP groups in reversed learning day 6.


Hit and CR rates laser on vs laser off, NpHR mice LEC injection, CA1 implant


P Hit = 0. 46 P CR = 0.16

NpHR=10

Hit and CR rates laser on vs laser off, EYFP mice LEC injection, CA1 implant

Mann-Whitney U-test

P Hit = 1.00 P CR = 0.30

EYFP=9

Hit and CR rates NpHR mice laser on vs EYFP mice laser on LEC injection, CA1 implant

Mann-Whitney U-test

P Hit = 0.25 P CR = 0.002

NpHR=10 EYFP=9

Hit and CR rates NpHR mice laser off vs EYFP mice laser off LEC injection, CA1 implant

Mann-Whitney U-test

P Hit = 0.53 P CR = 0.07

NpHR=10 EYFP=9

Hit and CR rates laser on vs laser off, NpHR mice LEC injection, DG implant

Mann-Whitney U-test


NpHR=9

Hit and CR rates laser on vs laser off, EYFP mice LEC injection, DG implant

Mann-Whitney U-test

P Hit = 1.00 P CR = 0.13

EYFP=10

Hit and CR rates NpHR mice laser on vs EYFP mice laser on LEC injection, DG implant

Mann-Whitney U-test

P Hit = 1.00 P CR = 0.75

NpHR=9 EYFP=10

Hit and CR rates NpHR mice laser off vs EYFP mice laser off LEC injection, DG implant

Mann-Whitney U-test

P Hit = 1.00 P CR = 0.51

NpHR=9; EYFP=10;

Hit and CR rates laser on vs laser off, NpHR mice MEC injection, CA1 implant

Mann-Whitney U-test


NpHR=10

Hit and CR rates laser on vs laser off, EYFP mice MEC injection, CA1 implant

Mann-Whitney U-test

P Hit = 0.72 P CR = 0.23

EYFP=10

Hit and CR rates NpHR mice laser on vs EYFP mice laser on, MEC injection, CA1 implant

Mann-Whitney U-test

P Hit = 0.28 P CR = 0.21

NpHR=10 EYFP=10

Hit and CR rates NpHR mice laser off vs EYFP mice laser off, MEC injection, CA1 implant

Mann-Whitney U-test


NpHR=10 EYFP=10



Supplementary Table 14 Statistical results on behavioral performance in Arch+ and

Ai35 groups in the (reversed) learning day 6.


Performance correct rate laser on vs laser off Arch+ mice CA1 implant


P = 0.007

Arch+ =9;

Performance correct rate laser on vs laser off Ai35 mice CA1 implant

Mann-Whitney U-test


Ai35 =9;

Performance correct rate Arch+ mice laser on vs Ai35 mice laser on CA1 implant

Mann-Whitney U-test

P = 8.6×10-5

Arch+ =9; Ai35 =9;

Performance correct rate Arch+ mice laser off vs Ai35 mice laser off CA1 implant

Mann-Whitney U-test

P = 0.433

Arch+ =9; Ai35 =9;

Performance correct rate laser on vs laser off Arch+ mice DG implant

Mann-Whitney U-test


Arch+ =11;

Performance correct rate laser on vs laser off Ai35 mice DG implant

Mann-Whitney U-test


Ai35 =10;

Performance correct rate Arch+ mice laser on vs Ai35 mice laser on DG implant

Mann-Whitney U-test

P = 0.268

Arch+ =11; Ai35 =10;

Performance correct rate Arch+ mice laser off vs Ai35mice laser off DG implant

Mann-Whitney U-test

P = 0.338

Arch+ =11; Ai35 =10;

It refers to the data shown in (Supplementary Fig. 12a4-5).


Supplementary Table 15 Statistical results on hit rate and correct rejection (CR)

rates in Arch+ and Ai35 groups in the (reversed) learning day 6.



Hit and CR rates laser on vs laser off Arch+ mice CA1 implant


P Hit = 0.788 P CR = 0.022

Arch+ =9;

Hit and CR rates laser on vs laser off Ai35 mice CA1 implant

Mann-Whitney U-test

P Hit = 0.180 P CR = 0.062

Ai35 =9

Hit and CR rates Arch+ mice laser on vs Ai35mice laser on CA1 implant

Mann-Whitney U-test

P Hit = 0.669 P CR = 4.1×10-4

Arch+=9; Ai35 =9

Hit and CR rates Arch+ mice laser off vs Ai35 mice laser off CA1 implant

Mann-Whitney U-test

P Hit = 0.88 P CR = 0.62

Arch+=9; Ai35 =9

Hit and CR rates laser on vs laser off Arch+ mice DG implant

Mann-Whitney U-test


Arch+ =11;

Hit and CR rates laser on vs laser off Ai35 mice, DG implant

Mann-Whitney U-test


Ai35 =10;

Hit and CR rates Arch+ mice laser on vs Ai35 mice laser on, DG implant

Mann-Whitney U-test

P Hit = 0.670 P CR = 0.189

Arch+ =11; Ai35 =10

Hit and CR rates Arch+ mice laser off vs Ai35 mice laser off, DG implant

Mann-Whitney U-test

P Hit = 0.064 P CR = 0.094

Arch+ =11; Ai35 =10



Supplementary Table 16 Number of recorded dCA1 PCs exhibiting different patterns

of spiking responses evoked by odor A and B.

Differential spiking responses to odor A and odor B

(go trials) (no-go trials)

Number of Calb(+) PC

(n =46 in total)

Number of Calb(-)PC

(n=232 in total)

Statistic test results

+ unresponsive 9 18 P = 0.022 *

unresponsive - 2 15 P = 0.75

+ - 4 3 P = 0.026 *

unresponsive + 3 16 P = 1.

- unresponsive 10 60 P = 0.71

- + 3 4 P = 0.92

unresponsive unresponsive 5 75 P = 0.0023 **

+ + 2 12 P = 1

- - 8 29 P = 0.35

It corresponds to the sample odor responses recorded from dCA1 PCs shown in Supplementary Fig. 14. Cells,

which showed odor-evoked increase (+) or decrease (-) in the spike rate (averaged in 1.5 s odor sample duration)

One-way ANOVA, P < 0.01, were regarded as responsive ones, other are unresponsive. Fisher's Exact test is used to

measure significance of the difference in the cell numbers between groups. n.s.: no significance.


nature neuroscience: doi:10.1038/nncpc in the distal, medial and proximal part of dca1. nature...

Documents