Deciphering the ancestral mechanisms of neural regeneration through single-cell analyses of jellyfish rhopalium repair.
The 3 matches · 1 of them tie a paragraph to a whole file, not to given lines: a weak match, whose lines are not tinted
- [1] § MATERIALS AND METHODS › Cross-species analysis ↔ code.zip/code/blastp.sh, the whole file · a weak match · score 0.77 · Anolis carolinensis, SAMap, Schmidtea mediterranea, lizard, cross species, rerio
- [2] § MATERIALS AND METHODS › Cross-species analysis ↔ code.zip/code/CrossSpecies_main.R, lines 144–213 · score 0.58 · Schmidtea mediterranea, cross species integrative, rerio, Xenopus, mapped, cell
- [3] § MATERIALS AND METHODS › Data quality control, integration, and analysis ↔ code.zip/code/CrossSpecies_main.R, lines 215–280 · score 0.56 · FindClusters, FindNeighbors, Seurat, RNA
Paper
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The authors' code
R · 280 lines · 12 KB · CC-BY-4.0 · 2 matches
- # ==============================================================================
- # Single-Cell RNA-seq Analysis Pipeline for Aurelia
- # Main Tasks:
- # 1. Subset cell clusters
- # 2. Cell stemness analysis (CytoTRACE2)
- # 3. Lineage/trajectory inference (Slingshot, Monocle3)
- # 4. Cross-species gene mapping & integration
- # 5. Visualization of UMAP, trajectories, and gene expression
- # ==============================================================================
- # Load required libraries
- library(Seurat)
- library(tidyverse)
- library(CytoTRACE2)
- library(SeuratDisk)
- library(slop)
- library(monocle3)
- library(writexl)
- library(rtracklayer)
- library(slingshot)
- ZS_nosplit <- read_rds('~/Aurelia/ZS_nosplit.rds')
- ZS_nosplit_sub <- subset(ZS_nosplit, RNA_snn_res.0.6 %in%
- c(0,1,4,7,8,10,17,18,21,22,28))
- set.seed(1234)
- ZS_nosplit_down <- ZS_nosplit[, sample(1:ncol(ZS_nosplit), 10000)]
- ZS_nosplit_down[['RNA']] <- as(ZS_nosplit_down[['RNA']], Class = 'Assay')
- write_rds(ZS_nosplit_down, '~/Aurelia/ZS_nosplit_down.rds')
- # ==============================================================================
- #Lineage Inference with Slingshot
- # ==============================================================================
- pdf('~/Aurelia/lineage3.pdf', width = 8, height = 8)
- # Run Slingshot trajectory analysis
- ZS_nosplit_slingshot <- slop::RunSlingshot(
- srt = subset(ZS_nosplit_down, RNA_snn_res.0.6 %in%
- c(0,1,4,7,8,10,17,18,21,22,28)),
- group.by = "RNA_snn_res.0.6",
- start = 1,
- reduction = "umap"
- )
- dev.off()
- # Visualize lineages on UMAP
- slop::CellDimPlot(
- ZS_nosplit_slingshot,
- group.by = "RNA_snn_res.0.6",
- reduction = "umap",
- lineages = paste0("Lineage", 1:3),
- lineages_span = 0.1,
- lineages_trim = c(0.05, 0.95)
- )
- # Plot pseudotime for each lineage
- umap_limits <- list(
- x = range(ZS_nosplit@reductions$[email hidden][,1]),
- y = range(ZS_nosplit@reductions$[email hidden][,2])
- )
- p1 <- FeatureDimPlot(ZS_nosplit_slingshot, features = "Lineage1",
- reduction = "umap", pt.size = 0.5) +
- xlim(umap_limits$x) + ylim(umap_limits$y)
- p2 <- FeatureDimPlot(ZS_nosplit_slingshot, features = "Lineage2",
- reduction = "umap", pt.size = 0.5) +
- xlim(umap_limits$x) + ylim(umap_limits$y)
- p3 <- FeatureDimPlot(ZS_nosplit_slingshot, features = "Lineage3",
- reduction = "umap", pt.size = 0.5) +
- xlim(umap_limits$x) + ylim(umap_limits$y)
- # Combine and save lineage plots
- pdf('~/Aurelia/reversion/lineage_all.pdf', width = 16, height = 5)
- p1 + p2 + p3 +p4
- dev.off()
- # ==============================================================================
- # Re-process Subsetted Data (Normalization, PCA, UMAP)
- # ==============================================================================
- ZS_nosplit_sub <- JoinLayers(ZS_nosplit_sub)
- sceobj_exp <- LayerData(ZS_nosplit_sub, assay = "RNA", layer = 'counts')
- # Create new Seurat object
- sceobj_new <- CreateSeuratObject(counts = sceobj_exp,
- meta.data = [email hidden])
- #Split assay by sample for integration
- sceobj_new[["RNA"]] <- split(sceobj_new[["RNA"]], f = sceobj_new$orig.ident)
- # ==============================================================================
- #: Export Cluster2 for Cross-Species Analysis
- # ==============================================================================
- C2 <- read_rds('~/Aurelia/reversion/cluster2.rds')
- C2[['RNA']] <- as(C2[['RNA']], Class = 'Assay')
- # Modify gene names to avoid conflicts
- C2_exp <- C2@assays$RNA@counts
- rownames(C2_exp) <- paste0(stringr::str_replace_all(rownames(C2_exp), 'gene', 'rna'), '.1')
- # Create annotated Seurat object
- C2 <- CreateSeuratObject(counts = C2_exp, meta.data = [email hidden])
- C2 <- NormalizeData(C2)
- C2 <- ScaleData(C2)
- C2$celltype <- 'C2'
- # Export as h5Seurat and h5ad (for Python/scanpy)
- SeuratDisk::SaveH5Seurat(C2, filename = "~/Aurelia/reversion/C2_ann.h5Seurat")
- SeuratDisk::Convert("~/Aurelia/reversion/C2_ann.h5Seurat", dest = "h5ad")
- write_rds(C2, '~/Aurelia/reversion/C2.rds')
- # ==============================================================================
- # Cross-Species Gene Mapping Functions
- # ==============================================================================
- trans_species_obj <- function(query_obj, subject_obj, map_gene) {
- subject_exp <- subject_obj@assays$RNA$counts
- subject_exp_duplicate_new <- subject_exp
- for (i in 1:5) {
- temp_exp <- subject_exp
- rownames(temp_exp) <- paste0(rownames(subject_exp), "-", i)
- subject_exp_duplicate_new <- rbind(temp_exp, subject_exp_duplicate_new)
- }
- map_gene <- map_gene[map_gene$subject_id %in% rownames(subject_exp_duplicate_new), ]
- subject_exp_duplicate_new <- subject_exp_duplicate_new[map_gene$subject_id, ]
- rownames(subject_exp_duplicate_new) <- map_gene$query_id
- subject_obj_new <- CreateSeuratObject(counts = subject_exp_duplicate_new, meta.data = [email hidden])
- merge_obj <- merge(query_obj, subject_obj_new)
- return(merge_obj)
- }
- trans_gene_species <- function(species1_vs_species2, species2_vs_species1) {
- colnames(species1_vs_species2) <- c(
- "query_id", "subject_id", "percent_identity", "alignment_length",
- "mismatch_count", "gap_open_count", "q_start", "q_end",
- "s_start", "s_end", "e_value", "bit_score"
- )
- colnames(species2_vs_species1) <- colnames(species1_vs_species2)
- best_hits_species1 <- species1_vs_species2 %>%
- group_by(query_id) %>%
- top_n(4, bit_score) %>%
- ungroup()
- best_hits_species2 <- species2_vs_species1 %>%
- group_by(query_id) %>%
- top_n(4, bit_score) %>%
- ungroup()
- rbh <- inner_join(best_hits_species1, best_hits_species2,
- by = c("query_id" = "subject_id", "subject_id" = "query_id")) %>%
- select(query_id, subject_id)
- return(rbh)
- }
- # ==============================================================================
- # Cross-Species Integration
- # ==============================================================================
- # Xenopus
- ROCs_sce <- SeuratDisk::LoadH5Seurat("~/Aurelia/regen/Xenopus/ROCs_sce.h5Seurat")
- Au_to_Xp <- read_table('~/Aurelia/regen/maps/XeAu/Au_to_Xe.txt', col_names = F)
- Xp_to_Au <- read_table('~/Aurelia/regen/maps/XeAu/Xe_to_Au.txt', col_names = F)
- map_gene_Au_Xp <- trans_gene_species(Au_to_Xp, Xp_to_Au)
- ROCs_sce$celltype <- 'Xenopus_ROCs'
- ROCs_sce$species <- 'Xenopus'
- # Hydra
- Hy_iCell <- SeuratDisk::LoadH5Seurat("~/Aurelia/regen/Hydra/Hy_iCell.h5Seurat")
- Au_to_Hy <- read_table('~/Aurelia/regen/maps/HyAu/Au_to_Hy.txt', col_names = F)
- Hy_to_Au <- read_table('~/Aurelia/regen/maps/HyAu/Hy_to_Au.txt', col_names = F)
- map_gene_Au_Hy <- trans_gene_species(Au_to_Hy, Hy_to_Au)
- Hy_iCell$celltype <- 'Hydra_iCell'
- Hy_iCell$species <- 'Hydra'
- # Schmidtea mediterranea
- Sc_neoblast <- SeuratDisk::LoadH5Seurat("~/Aurelia/regen/Schmidtea_mediterranea/Sc_neoblast.h5Seurat")
- Au_to_Sc <- read_table('~/Aurelia/regen/maps/ScAu/Au_to_Sc.txt', col_names = F)
- Sc_to_Au <- read_table('~/Aurelia/regen/maps/ScAu/Sc_to_Au.txt', col_names = F)
- map_gene_Au_Sc <- trans_gene_species(Au_to_Sc, Sc_to_Au)
- Sc_neoblast$celltype <- 'Sc_neoblast'
- Sc_neoblast$species <- 'Schmidtea_mediterranea'
- # Danio rerio
- Danio_rerio_sce_Me <- SeuratDisk::LoadH5Seurat("~/Aurelia/regen/Danio_rerio/Danio_re_Mesenchymal.h5Seurat")
- Au_to_Da <- read_table('~/Aurelia/regen/maps/DaAu/Au_to_Da.txt', col_names = F)
- Da_to_Au <- read_table('~/Aurelia/regen/maps/DaAu/Da_to_Au.txt', col_names = F)
- map_gene_Au_Da <- trans_gene_species(Au_to_Da, Da_to_Au)
- Danio_rerio_sce_Me$celltype <- 'Danio_rerio_Mesenchymal'
- Danio_rerio_sce_Me$species <- 'Danio_rerio'
- # Acoel
- Ac_neoblast <- SeuratDisk::LoadH5Seurat("~/Aurelia/regen/Acoel/Ac_neoblast1.h5Seurat")
- Au_to_Ac <- read_table('~/Aurelia/regen/maps/AcAu/Au_to_Ac.txt', col_names = F)
- Ac_to_Au <- read_table('~/Aurelia/regen/maps/AcAu/Ac_to_Au.txt', col_names = F)
- map_gene_Au_Ac <- trans_gene_species(Au_to_Ac, Ac_to_Au)
- Ac_neoblast$celltype <- 'Ac_neoblast'
- Ac_neoblast$species <- 'Acoel'
- # ==============================================================================
- # Merge All Species and Integration
- # ==============================================================================
- query_exp <- C2@assays$RNA$counts
- query_exp_duplicate <- query_exp
- query_exp_duplicate_new <- query_exp
- for (i in 1:5) {
- row.names(query_exp_duplicate) <- paste0(row.names(query_exp), "_", i)
- query_exp_duplicate_new <- rbind(query_exp_duplicate,query_exp_duplicate_new)
- }
- identical(colnames(query_exp_duplicate_new),
- row.names([email hidden]))
- query_obj <- CreateSeuratObject(counts = query_exp_duplicate_new,
- meta.data = [email hidden])
- trans_Au_Ac_obj <- trans_species_obj(query_obj = query_obj,
- subject_obj = Ac_neoblast,
- map_gene = map_gene_Au_Ac)
- trans_Au_Da_obj <- trans_species_obj(query_obj = query_obj,
- subject_obj = Danio_rerio_sce_Me,
- map_gene = map_gene_Au_Da)
- trans_Au_Hy_obj <- trans_species_obj(query_obj = query_obj,
- subject_obj = Hy_iCell,
- map_gene = map_gene_Au_Hy)
- trans_Au_Xe_obj <- trans_species_obj(query_obj = query_obj,
- subject_obj = ROCs_sce,
- map_gene = map_gene_Au_Xp)
- trans_Au_Sc_obj <- trans_species_obj(query_obj = query_obj,
- subject_obj = Sc_neoblast,
- map_gene = map_gene_Au_Sc)
- trans_Au_La_obj <- trans_species_obj(query_obj = query_obj,
- subject_obj = Sc_neoblast,
- map_gene = map_gene_Au_Sc)
- C2$species <- 'AU'
- merge_all_species <- merge(C2,
- list(trans_Au_Ac_obj,
- trans_Au_Da_obj,
- trans_Au_Hy_obj,
- trans_Au_Xe_obj,
- trans_Au_Sc_obj))
- merge_all_species <- JoinLayers(merge_all_species)
- merge_all_species <- NormalizeData(merge_all_species) %>% ScaleData()
- Idents(merge_all_species) <- 'species'
- merge_all_species_exp <- AverageExpression(merge_all_species,assays = 'RNA')$RNA
- write.csv(merge_all_species_exp,'~/Aurelia/regen/merge_all_species_exp.csv')
- write_rds(merge_all_species,file = '~/Aurelia/regen/merge_all_species.rds')
- # ==============================================================================
- # Integration (Harmony, CCA, RPCA)
- # ==============================================================================
- sce <- read_rds('~/Aurelia/regen/merge_all_species.rds')
- sce <- JoinLayers(sce)
- sce <- CreateSeuratObject(sce@assays$RNA, meta.data = [email hidden])
- sce[["RNA"]] <- split(sce[["RNA"]], f = sce$species)
- sce <- NormalizeData(sce)
- sce <- FindVariableFeatures(sce)
- sce <- ScaleData(sce)
- sce <- RunPCA(sce)
- # Harmony
- sce <- IntegrateLayers(object = sce, method = HarmonyIntegration,
- orig.reduction = "pca", new.reduction = "integrated.harmony")
- sce <- RunUMAP(sce, reduction = "integrated.harmony", dims = 1:30, reduction.name = "umap.harmony")
- # CCA
- sce_cca <- IntegrateLayers(object = sce, method = CCAIntegration,
- orig.reduction = "pca", new.reduction = "integrated.cca")
- sce_cca <- RunUMAP(sce_cca, reduction = "integrated.cca", dims = 1:30, reduction.name = "umap.cca")
- # RPCA
- sce_rpca <- IntegrateLayers(object = sce, method = RPCAIntegration,
- orig.reduction = "pca", new.reduction = "integrated.rpca")
- sce_rpca <- RunUMAP(sce_rpca, reduction = "integrated.rpca", dims = 1:30, reduction.name = "umap.rpca")
- # Clustering
- sce <- FindNeighbors(sce, reduction = "integrated.harmony", dims = 1:30)
- sce <- FindClusters(sce, resolution = c(0.2, 0.5, 1))
- # Plot UMAP
- pdf('~/Aurelia/regen/merge_all_species_umap.pdf', width = 10, height = 10)
- DimPlot(sce, group.by = 'species')
- DimPlot(sce_cca, reduction = 'umap.cca', group.by = 'species')
- DimPlot(sce_rpca, reduction = 'umap.rpca', group.by = 'species')
- dev.off()
- write_rds(sce_cca, '~/Aurelia/regen/merge_all_species.rds')
- # Export results
- exp <- AverageExpression(sce)
- write_rds(exp$RNA, '~/Aurelia/regen/exp.rds')
- writexl::write_xlsx(
- list(map_gene_Au_Sc = map_gene_Au_Sc,
- map_gene_Au_Ac = map_gene_Au_Ac,
- map_gene_Au_Da = map_gene_Au_Da,
- map_gene_Au_Hy = map_gene_Au_Hy,
- map_gene_Au_Xp = map_gene_Au_Xp),
- path = '~/Aurelia/regen/all_species_mapping_gene.xlsx'
- )
- # ==============================================================================
CrossSpecies_main.R, under CC-BY-4.0 · at the source
Overview
- Muping Coastal Environment Research Station, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai, Shandong 264003, China
- University of Chinese Academy of Sciences, Beijing 100049, China
- College of Bio-Medicine and Health, Huazhong Agricultural University, Wuhan, Hubei 430070, China
- Department of Ocean Science, The Hong Kong University of Science and Technology, Hong Kong SAR, China
- School of Life Science, Yantai University, Yantai, Shandong 264005, China
Abstract
The rhopalium, an early specialized centralized nervous structure in medusozoans, exhibits a remarkable regenerative capacity after removal, offering a unique model for investigating the ancestral regulatory logic of neural regeneration. However, the cellular and molecular mechanisms underlying the process remain unknown. Here, we defined distinct stages of rhopalium regeneration in the jellyfish Aurelia coerulea and constructed a stage-specific cell landscape and gene regulatory networks. Notably, we identified a population of wound-induced cells that emerges rapidly postinjury and activates Wnt signaling to initiate blastema formation; cross-species comparisons revealed that this regulatory mechanism is conserved across metazoans. We further identified that retinoic acid signaling is essential for photoreceptor regeneration and ocelli reconstruction. The findings presented herein uncover the ancestral regulatory mechanisms governing neural regeneration in the early centralized nervous system, providing insights into the common evolutionary origins of neural repair across organisms.
Reproduced under the paper's license (CC BY-NC), from the paper cited above.
Repository
Its files are read in the Code ↔ Paper reader above, with 3 matches between paragraphs and lines of code.
Zenodo 19466460
Availability: 1 check, the latest on 28 September 2026: the link answers (HTTP 200)
- 28 September 2026: the link answers (HTTP 200)
2 files
- code.zip/
code/ — R, 280 lines, 2 matchesCrossSpecies_main.R - code.zip/
code/ — Shell, 51 lines, 1 matchblastp.sh
The paper's code and data availability statement is in the Data section.
Tracing map
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What the map holds:
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- neither the text of the paper nor the code itself.
Its JSON (tracing-map.json) is deposited on Zenodo with its DOI once the map is validated.
Data
No dataset and no data link were found in the paper.
Data, code, and materials availability
Sequencing data have been deposited in the NCBI under BioProject accession number PRJNA1224934. All other data needed to evaluate and reproduce the results in the paper are present in the paper and/
Reproduced under the paper's license (CC BY-NC), from the paper cited above.
Versions
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Version 1, 28 September 2026: the first record
Recorded: type, language, journal, volume, issue, pages, dates, 12 authors, 8 MeSH terms, 72 references.
Cite
This paper
Li, Y., Wang, F., Sun, T., Ma, X., Yu, Z., Xu, Z., Wang, W., Xing, Y., Peng, S., Wang, L., Zhao, J., & Dong, Z. (2026). Deciphering the ancestral mechanisms of neural regeneration through single-cell analyses of jellyfish rhopalium repair. Science advances, 12(21), eaeb8034. https://
BibTeX
@article{li2026decipheri
author = {Li, Yongxue and Wang, Fanghan and Sun, Tingting and Ma, Xiaoran and Yu, Zhangyi and Xu, Ziyue and Wang, Wenhui and Xing, Yixuan and Peng, Saijun and Wang, Lei and Zhao, Jianmin and Dong, Zhijun},
title = {{Deciphering the ancestral mechanisms of neural regeneration through single-cell analyses of jellyfish rhopalium repair}},
journal = {Science advances},
year = {2026},
month = may,
volume = {12},
number = {21},
pages = {eaeb8034},
publisher = {American Association for the Advancement of Science},
issn = {2375-2548},
doi = {10.1126/
url = {https://
pmid = {42160431},
pmcid = {PMC13189125}
}
RIS
TY - JOUR
AU - Li, Yongxue
AU - Wang, Fanghan
AU - Sun, Tingting
AU - Ma, Xiaoran
AU - Yu, Zhangyi
AU - Xu, Ziyue
AU - Wang, Wenhui
AU - Xing, Yixuan
AU - Peng, Saijun
AU - Wang, Lei
AU - Zhao, Jianmin
AU - Dong, Zhijun
TI - Deciphering the ancestral mechanisms of neural regeneration through single-cell analyses of jellyfish rhopalium repair
T2 - Science advances
J2 - Sci Adv
PY - 2026
DA - 2026/
VL - 12
IS - 21
SP - eaeb8034
SN - 2375-2548
PB - American Association for the Advancement of Science
DO - 10.1126/
UR - https://
LA - en
ER -
CSL-JSON
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"title": "Deciphering the ancestral mechanisms of neural regeneration through single-cell analyses of jellyfish rhopalium repair",
"container-title": "Science advances",
"author": [
{
"family": "Li",
"given": "Yongxue"
},
{
"family": "Wang",
"given": "Fanghan"
},
{
"family": "Sun",
"given": "Tingting"
},
{
"family": "Ma",
"given": "Xiaoran"
},
{
"family": "Yu",
"given": "Zhangyi"
},
{
"family": "Xu",
"given": "Ziyue"
},
{
"family": "Wang",
"given": "Wenhui"
},
{
"family": "Xing",
"given": "Yixuan"
},
{
"family": "Peng",
"given": "Saijun"
},
{
"family": "Wang",
"given": "Lei"
},
{
"family": "Zhao",
"given": "Jianmin"
},
{
"family": "Dong",
"given": "Zhijun"
}
],
"container-title-short":
"volume": "12",
"issue": "21",
"page": "eaeb8034",
"DOI": "10.1126/
"PMID": "42160431",
"PMCID": "PMC13189125",
"ISSN": "2375-2548",
"publisher": "American Association for the Advancement of Science",
"URL": "https://
"language": "en",
"issued": {
"date-parts": [
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]
}
}
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You validate the map as this page shows it: 1 repository of the authors' code, each at its verified commit and with its license, 2 scripts, and 3 matches between paragraphs and code (see the Code and Map sections). It then receives a DOI on Zenodo, with you (your ORCID iD) and OSCR as its creators; the code itself is not deposited.
The map's fingerprint: sha256:9e3d1ac6b66ec601…
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The badge links the code to this page. Copy one of these into the README of the paper's code: only you decide where it goes, and nothing is changed for you.
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Discussion, reproductions, activity
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Reproductions: reports from readers who ran the authors' code: what they reproduced, with which environment, commit and data. It opens with sign-in.
Activity: what happens around this paper: new versions of its record, its map's validation, discussions and reproductions. It opens with sign-in.
