1
nature research | reporting summary
April 2018
Corresponding author(s):
Yuki Oka
Reporting Summary
Nature Research wishes to improve the reproducibility of the work that we publish. This form provides structure for consistency
and transparency
in reporting. For further information on Nature Research policies, see
Authors & Referees
and the
Editorial Policy Checklist
.
Statistical parameters
When statistical analyses are reported, confirm that the following items are present in the relevant location (e.g. figure lege
nd, table legend, main
text, or Methods section).
n/a
Confirmed
The exact sample size (
n
) for each experimental group/condition, given as a discrete number and unit of measurement
An indication of whether measurements were taken from distinct samples or whether the same sample was measured repeatedly
The statistical test(s) used AND whether they are one- or two-sided
Only common tests should be described solely by name; describe more complex techniques in the Methods section.
A description of all covariates tested
A description of any assumptions or corrections, such as tests of normality and adjustment for multiple comparisons
A full description of the statistics including central tendency (e.g. means) or other basic estimates (e.g. regression coeffici
ent) AND
variation (e.g. standard deviation) or associated estimates of uncertainty (e.g. confidence intervals)
For null hypothesis testing, the test statistic (e.g.
F
,
t
,
r
) with confidence intervals, effect sizes, degrees of freedom and
P
value noted
Give P values as exact values whenever suitable.
For Bayesian analysis, information on the choice of priors and Markov chain Monte Carlo settings
For hierarchical and complex designs, identification of the appropriate level for tests and full reporting of outcomes
Estimates of effect sizes (e.g. Cohen's
d
, Pearson's
r
), indicating how they were calculated
Clearly defined error bars
State explicitly what error bars represent (e.g. SD, SE, CI)
Our web collection on
statistics for biologists
may be useful.
Software and code
Policy information about
availability of computer code
Data collection
Lickometer (Dialog Instruments), Leica Application Suite X 3.5.5.19976 and 10x Genomics Cell Ranger 3.0.1. were used for data c
ollection.
Data analysis
Custom R code, Custom Python 3 code and GraphPad Prism 8 were used to analyze data. Code is available from the corresponding
author upon reasonable request. ScRNA-seq data were processed in R 3.5.1 using the following analysis and data visualization pa
ckages:
Seurat v.3.0.3.9019, gplots 3.0.1.1, Hmisc 4.2-0, ggplot2 3.2.0, viridis 0.5.1., scales 1.0.0., spatstat 1.62-2, Matrix 1.2-14,
dplyr 0.8.3,
cowplot 1.0.0, psych 1.9.12.31.
For manuscripts utilizing custom algorithms or software that are central to the research but not yet described in published lit
erature, software must be made available to editors/reviewers
upon request. We strongly encourage code deposition in a community repository (e.g. GitHub). See the Nature Research
guidelines for submitting code & software
for further information.
2
nature research | reporting summary
April 2018
Data
Policy information about
availability of data
All manuscripts must include a
data availability statement
. This statement should provide the following information, where applicable:
- Accession codes, unique identifiers, or web links for publicly available datasets
- A list of figures that have associated raw data
- A description of any restrictions on data availability
The behavioral and histology data that support the findings are available from the corresponding author upon reasonable request
. Raw and fully processed scRNA-
seq data is available at the NCBI Gene Expression Omnibus (GEO accession # GSE154048).
Field-specific reporting
Please select the best fit for your research. If you are not sure, read the appropriate sections before making your selection.
Life sciences
Behavioural & social sciences
Ecological, evolutionary & environmental sciences
For a reference copy of the document with all sections, see
nature.com/authors/policies/ReportingSummary-flat.pdf
Life sciences study design
All studies must disclose on these points even when the disclosure is negative.
Sample size
No statistics to determine sample size were used. Sample size is similar to recent papers in
behavioural neuroscience (Augustine, V. et al. Nature 555, 204-209, 2018 and Lee, S. et al. Nature 568, 93-97, 2019)
Data exclusions
For behavioral experiments, all data were included after verification of virus expression in the target brain area. For single
cell RNA-seq
experiments common thresholding to remove doublets and unhealthy cells was implemented.
Replication
We verified all experiments and analyses by replicating them by more than two lab
members. All of the replication attempts were successful.
Randomization
No randomization was used for behavioral assays. All solutions were presented in an alternate fashion. In our pilot studies, we
randomized the
order and found no difference in the results.
Blinding
No blinding was used for data collection. All data were analyzed using an automated software that calculates preference to each
solution.
Thus, randomization is less relevant in this study.
Reporting for specific materials, systems and methods
Materials & experimental systems
n/a
Involved in the study
Unique biological materials
Antibodies
Eukaryotic cell lines
Palaeontology
Animals and other organisms
Human research participants
Methods
n/a
Involved in the study
ChIP-seq
Flow cytometry
MRI-based neuroimaging
Antibodies
Antibodies used
rabbit anti-c-Fos (1:500, Cell Signaling, #2250), chicken anti-GFP (1:1000, Abcam, ab13970, Lot# GR3190550-1), rabbit-anti-ETV1
(1:500, Abcam, ab81086), sheep anti-Foxp2 (1:2000, R&D systems, AF5647, Lot# CCUB0109061), and rat anti-mCherry (1:500,
Invitrogen, 16D7). For secondary antibodies we used donkey-anti-rabbit-Cy3 (711-165-152, Lot# 144186), donkey-anti-rabbit-
A488 (711-545-152, Lot# 147626), donkey-anti-chicken-A488 (703-545-155, Lot# 147805), donkey-anti-sheep-A488
(713-545-147, Lot# 144916), donkey-anti-rat-Cy3 (712-165-150, Lot# 146156). All secondary antibodies were used at 1:500
dilution and were obtained from Jackson Immunoresearch.
Validation
All antibodies were puchased from vendors in the USA. These products are normally quality controlled at the company.
3
nature research | reporting summary
April 2018
Nevertheless, we characterized and validated their signals in the brain regions known to express these genes prior to
experiments.
Animals and other organisms
Policy information about
studies involving animals
;
ARRIVE guidelines
recommended for reporting animal research
Laboratory animals
The following mice were purchased from the Jackson Laboratory: C57BL/6J, stock number 00064; Ai14, stock number 007914;
Ai3, stock number 007903. Rxfp1-P2A-Cre line was prepared in J. Ngai lab (UC Berkeley). Trap2 mice were a gift from Liqun Luo
(Stanford University). PDYN-Cre mice were provided by B. Lowell (Harvard Medical School) and M. Krashes (NIH).
Wild animals
No wild animals were used in the study.
Field-collected samples
No field collected samples were used in the study.