Published May 18, 2022 | Version v3
Journal article Open

Orexin neurons inhibit sleep to promote arousal

  • 1. Department of Neurology, Division of Sleep Medicine, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA, 02215, USA
  • 2. Department of Medicine, Division of Endocrinology, Diabetes and Metabolism. Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA, USA
  • 3. Department of Neurological Surgery, University of California Davis School of Medicine, Davis, CA, USA.
  • 4. Department of Pediatrics, Division of Endocrinology, F.M. Kirby Neurobiology Center. Children's Hospital and Harvard Medical School, Boston, MA, USA.
  • 5. Department of Biology, Graduate School of Science and Engineering. University of Toyama, Toyama, Japan.
  • 6. Department of Molecular Biology, Massachusetts General Hospital, Boston, MA, 02114, USA

Description

OVERVIEW
The following README contains: i) the instruction to set up an R environment (R version 3.6.3) and to install R Studio on a Windows/macOS operative system; ii) the instructions to install Seurat 3.1.5 along with all its dependencies; iii) the software code used to generate all the plots and figure as showed in the manuscript.

SYSTEM REQUIREMENTS
The following analysis can be performed on a standard computer with sufficient RAM to support the in-memory operation required by Seurat 3.1.5. The installation of Seurat 3.1.5 and all its dependencies have been tested on both Windows (version 10 64-bit ) and macOS (Big Sur version 11.6.1) operating systems. 

ALL PACKAGES and DEPENDENCIES
attached base packages:
[1] stats     graphics  grDevices utils     datasets  methods   base    
other attached packages:
[1] biomaRt_2.42.1 Seurat_3.1.5   Matrix_1.4-0  

  [1] TH.data_1.1-0        Rtsne_0.15           colorspace_2.0-2     ellipsis_0.3.2       ggridges_0.5.3       leiden_0.3.9        
  [7] listenv_0.8.0        remotes_2.4.2        ggrepel_0.9.1        bit64_4.0.5          AnnotationDbi_1.48.0 fansi_1.0.2        
 [13] mvtnorm_1.1-3        mathjaxr_1.4-0       codetools_0.2-18     splines_3.6.3        mnormt_2.0.2         cachem_1.0.6        
 [19] TFisher_0.2.0        jsonlite_1.7.3       ica_1.0-2            dbplyr_2.1.1         cluster_2.1.2        png_0.1-7          
 [25] uwot_0.1.11          sctransform_0.3.3    BiocManager_1.30.16  compiler_3.6.3       httr_1.4.2           assertthat_0.2.1    
 [31] fastmap_1.1.0        lazyeval_0.2.2       cli_3.1.1            prettyunits_1.1.1    htmltools_0.5.2      tools_3.6.3        
 [37] rsvd_1.0.5           igraph_1.2.11        gtable_0.3.0         glue_1.6.1           RANN_2.6.1           reshape2_1.4.4      
 [43] dplyr_1.0.7          rappdirs_0.3.3       Rcpp_1.0.8           Biobase_2.46.0       vctrs_0.3.8          multtest_2.42.0    
 [49] ape_5.6-1            nlme_3.1-155         lmtest_0.9-39        stringr_1.4.0        globals_0.14.0       rbibutils_2.2.7    
 [55] lifecycle_1.0.1      irlba_2.3.5          XML_3.99-0.3         future_1.23.0        MASS_7.3-55          zoo_1.8-9          
 [61] scales_1.1.1         hms_1.1.1            parallel_3.6.3       sandwich_3.0-1       RColorBrewer_1.1-2   curl_4.3.2          
 [67] memoise_2.0.1        reticulate_1.24      pbapply_1.5-0        gridExtra_2.3        ggplot2_3.3.5        stringi_1.7.6      
 [73] RSQLite_2.2.7        mutoss_0.1-12        S4Vectors_0.24.4     plotrix_3.8-2        BiocGenerics_0.32.0  Rdpack_2.1.3        
 [79] rlang_1.0.1          pkgconfig_2.0.3      matrixStats_0.61.0   lattice_0.20-45      ROCR_1.0-11          purrr_0.3.4        
 [85] patchwork_1.1.1      htmlwidgets_1.5.4    cowplot_1.1.1        bit_4.0.4            tidyselect_1.1.1     parallelly_1.30.0  
 [91] RcppAnnoy_0.0.19     plyr_1.8.6           magrittr_2.0.2       R6_2.5.1             IRanges_2.20.2       generics_0.1.2      
 [97] multcomp_1.4-18      DBI_1.1.2            pillar_1.7.0         sn_2.0.1             fitdistrplus_1.1-6   survival_3.2-13    
[103] tibble_3.1.6         future.apply_1.8.1   tsne_0.1-3           crayon_1.4.2         KernSmooth_2.23-20   utf8_1.2.2          
[109] BiocFileCache_1.10.2 tmvnsim_1.0-2        plotly_4.10.0        progress_1.2.2       grid_3.6.3           data.table_1.14.2  
[115] blob_1.2.2           metap_1.7            digest_0.6.29        tidyr_1.2.0          numDeriv_2016.8-1.1  openssl_1.4.6      
[121] stats4_3.6.3         munsell_0.5.0        viridisLite_0.4.0    askpass_1.1   


SOFTWARE INSTALLATION GUIDE
1.    R version 3.6.3 for Windows and for macOS can be downloaded at: https://cran.r-project.org/bin/windows/base/old/3.6.3/ (Windows) and https://cran.r-project.org/bin/macosx/ (macOS). 
2.    Packages “remotes”, “BiocManager”, “biomaRt”, “Matrix” can be installed running the command: install.packages() from R console. 
3.    Seurat 3.1.5 and all its dependencies can be installed running the following command line: remotes::install_version("Seurat", version = "3.1.5")
4.    A typical installation time on a “normal” desktop computer is around 15-20 minutes. 

INSTRUCTION FOR USE/EXPECTED OUTPUT/EXPECTED RUN TIME
All the instructions to install and run the code on the data provided in this folder are in the document “script_NC.txt”. The expected outputs of the code are: 1) Supplementary tables 2-7; 2) Figure 5 (A-D); 3) Supplementary figures 9-11.
Expected run time is about 45 min for all the analyses. 

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