zebrafish fin regeneration

Our results suggest for the first time a role for Mmp13 in the regulation of the regeneration processes in zebrafish fin. The zebrafish as a model for complex tissue regeneration. The closest TSS per DAR was chosen by using BEDTools [60] version 2.27.1 and Ensembl gene annotation (release 85). 2009;460:60–5. statement and Cancel Unsubscribe. The fourth fin ray counting from the ventral side was used for consistency. Growth control in the ontogenetic and regenerating zebrafish fin. Integrating transcriptomes and chromatin accessibility maps, we identified thousands of novel regeneration enhancer elements and validated the enhancer activity of a few dozen candidates. HJL and YH performed the experiments. Average ATAC-seq signals were plotted on top of each heatmap (line plots). The unmethylated lambda DNA was used to calculate the bisulfite conversion rate. Vitamin D is an essential nutrient that has long been known to regulate skeletal growth and integrity. 2008;24:525–49. We found that 26% of ATAC peaks were in promoters (2 kb regions around transcription start site (TSS)), while 45% of ATAC peaks were distal (> 10 kb) to TSS (Additional file 1: Figure S4b). A number of genes have been implicated in regeneration, but the regulation of these genes, particularly pertaining to regeneration in higher vertebrates, remains an interesting and mostly open question. Differentially expressed genes during fin regeneration. Katsuyama T, Paro R. Epigenetic reprogramming during tissue regeneration. The datasets generated and analyzed during the current study are available in the NCBI’s Gene Expression Omnibus (GEO) repository (ATAC-seq: GSE126700 [69], RNA-seq: GSE126701 [70], and WGBS: GSE126702 [71]). Twitter handles: Hyung Joo Lee @hyungjoo_lee. Then Tn5 insertion events from ATAC-seq in 200 bp windows around these motif sites were counted. The reads were de-multiplexed by using sample-specific index sequences. Heatmaps and average line plots of DNA methylation and ATAC-seq signal levels of DARs along with their neighboring regions were plotted by using deepTools [55]. COVID-19 is an emerging, rapidly evolving situation. The right pectoral fin inner-vation was surgically removed before amputation, while 2019. https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE126702. Zebrafish caudal fins were amputated at 50% of their original length using razor blades. In addition, the vast majority (> 99%) of all the lowly methylated regions in sp7+ cells displayed little change in DNA methylation during regeneration (Additional file 1: Figure S2j). Mouse digit tip regeneration is mediated by fate-restricted progenitor cells. Only 30 (1.0%) of sp7+ cell-specific hypoDMRs were predicted as potential regeneration-specific DMRs. Dev Genes Evol. Over the past two decades of intense study, significant advances have been made in identifying both the regenerative cell sources and molecular signaling pathways in a variety of organs in adult zebrafish. e Representative pictures of the fin regenerate from the mutant (fosl1atw4/tw4) and their wildtype (fosl1a+/+) littermates at 2 dpa. Rossi A, Kontarakis Z, Gerri C, Nolte H, Hölper S, Krüger M, et al. Lee, H.J., Hou, Y., Chen, Y. et al. Nat Methods. Then, the expression levels of the zebrafish ortholog genes corresponding to those TFs were examined. 4d, e). This suggests that the small number of regeneration-specific DMRs were most likely false positives. Clipboard, Search History, and several other advanced features are temporarily unavailable. The skeleton of adult zebrafish fins comprises lepidotrichia, which are dermal bones of the rays, and actinotrichia, which are non-mineralized spicules at the distal margin of the appendage. Their regeneration processes were photographed daily up to 5 dpa. Goldman JA, Kuzu G, Lee N, Karasik J, Gemberling M, Foglia MJ, et al. HJL, YH, TW, and SLJ interpreted the data. Get the latest public health information from CDC: https://www.coronavirus.gov, Get the latest research information from NIH: https://www.nih.gov/coronavirus, Find NCBI SARS-CoV-2 literature, sequence, and clinical content: https://www.ncbi.nlm.nih.gov/sars-cov-2/. 2013;10:1213–8. For DNA extraction, 15 uninjured fins and 60 blastema per replicate were collected from transgenic zebrafish Tg(sp7:EGFP) and sp7+ and sp7− cell populations were isolated by using FACS as described above. Lee HJ, Hou Y, Chen Y, Dailey ZZ, Riddihough A, Jang HS, Wang T, Johnson SL. We would like thank Ali Wilkening and Kara Quaid for proofreading the manuscript. Abstract. Distal enhancer elements of the differentially expressed genes were defined as ATAC-seq peaks located closest to the TSS, but also farther than 10 kb, by using BEDTools [60] version 2.27.1. 2c), consistent with cellular processes characteristic of fin regeneration. 2006;299(2):438-454. **p < 0.01; Mann–Whitney U test. In Medaka, another teleost fish, a small-scale cDNA microarray screen was reported Recent work has shown the existence of tissue regeneration enhancer elements that trigger regenerative genetic program upon tissue damage in zebrafish [33]. Buscar en Biblos-e Archivo Esta colección. 2010;38:576–89. Similarly, genes that were upregulated or downregulated during regeneration exhibited increased and decreased ATAC-seq signals in their genomic neighborhood, respectively (Additional file 1: Figure S4g). 2012;9:357–9. The purified libraries were eluted with 20 μL of nuclease-free water. 2012;365:339–49. NOTE: The speed of zebrafish fin regeneration depends on the amount of resected fin tissue, i.e., the more fin tissue is resected, the faster the fin regenerates 13. Bioinformatics. To avoid any bias from the DMR calling algorithm, we tested a different algorithm and obtained similar results (Additional file 1: Figure S2f). Water temperature was maintained at 28.5 °C for animals. Signaling networks organizing regenerative growth of the zebrafish fin. Comparative aspects of animal regeneration. 1a; Additional file 2: Table S1; see below). 2015;524:230–3. This phenotype is replicated in four independent mutant alleles (Additional file 1: Figure S6c-e), confirming that mutation of fosl1a was causal for this regeneration defect. Mol Cell. It is noteworthy that we identified much fewer sp7− cell-specific hypoDMRs than sp7+ cell-specific hypoDMRs (Additional file 1: Figure S2g). Hirose K, Shimoda N, Kikuchi Y. The predicted target genes of Fra1 were not upregulated as highly in mutant fish as in wildtype at 1 dpa (left), consistent with the delayed regeneration phenotype. A blastema is a mass of proliferative and morphologically homogeneous cells that have the capability to grow and develop into a lost body part. The two-sided Wilcoxon signed-rank test was used to test statistical differences of gene expression changes in regenerates between mutant and wildtype zebrafish, by using the wilcox.test function with the parameter: paired = T. Sample sizes and p values are indicated in the figures or figure legends. 2005;310:1957–60. HJL was a McDonnell International Scholar (Washington University in St. Louis) and a Kwanjeong Educational Foundation Abroad Scholar (South Korea). Zebrafish caudal fin is a unique regeneration system to model the injury response and regeneration of vertebrate appendages despite being a simple structure without muscular and adipose tissues. Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. The IDR analysis was performed following ENCODE’s guidelines [65]. We examined a time-course expression of EGFP in regenerating fins and confirmed that EGFP expression was localized to the segmented bony fin rays in uninjured fin and strongly upregulated in the regenerates (Additional file 1: Figure S2a) [7, 29]. Dev Cell. d DARs with decreasing signals in sp7− cells during fin regeneration. Development. After amputation, the zebrafish caudal fin robustly regenerates via formation of blastema. Regeneration-specific gene activation with gain of chromatin accessibility. Curr Protoc Bioinformatics. In terminally differentiated cells, many developmental genes were repressed by epigenetic mechanism, such as DNA methylation over their cis-regulatory elements. Epub 2015 Apr 27. BEDTools: a flexible suite of utilities for comparing genomic features. After as little as one month, most of the missing tissue has been regenerated by cardiomyocytes.Regenerated heart muscles come from proliferation of differentiated cardiomyocytes. DNA methylome maps of sp7+ and sp7− cells during fin regeneration. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Low levels of DNA methylation over the DARs associated with regeneration genes in uninjured differentiated cells support a model that, in animals with high regeneration abilities, differentiated cells maintain a permissive epigenetic state (lowly methylated, yet low chromatin accessibility) over regulatory elements that are important for regeneration, thus might allow for rapid gene activation when needed for regeneration [40]. Zebrafish fin regeneration 475 signalling pathways. Regeneration enhancers identified in sp7+ cells drove GFP expression along the regenerating fin rays, while sp7− regeneration enhancers gave different patterns of GFP expression in the regenerates (Fig. Nucleic Acids Res. 4a; Additional file 1: Figure S6a). 1970;10:119–32. YC, ZZD, and AR assisted with the regeneration enhancer assay and mutant zebrafish generation. Epimorphic regeneration requires the presence or creation of pluripotent cells capable of reproducing lost organs. 2009;19:1044–56. fins; regeneration; tissue regeneration enhancer elements; vitamin D; zebrafish. These DMRs might reflect biological variation or stochasticity and do not have enough statistical confidence to be determined as regeneration-specific DMRs. The zebrafish fin provides a valuable model to study the epimorphic type of regeneration, whereby the amputated part of the appendage is nearly perfectly replaced. Asterisks indicate that F1 transgenic zebrafish line was established for a given enhancer element. For the regenerating blastema, caudal fins were allowed to regenerate at 28.5 °C for 4 days. Bioinformatics. In total, 18 out of 25 (72%) candidates displayed enhancer activities in regenerating fin while no activity was observed from 9 negative controls (Table 1). All sequencing data processed in the current study have been visualized in the WashU Epigenome Browser and is publicly available through the following url: https://epigenome.wustl.edu/FinRegen/. Molecular mechanisms of tissue regeneration have been studied mostly in the context of gene expression and function, and much remains to be investigated to decipher the gene regulatory networks involved in regeneration. Jopling C, Sleep E, Raya M, Martí M, Raya A, Izpisúa Belmonte JC. fgf20 is essential for initiating zebrafish fin regeneration. A total of 99.5 ng of genomic DNA per replicate, together with 0.5 ng of unmethylated lambda DNA (Promega), was treated with bisulfite and cleaned by using the EZ DNA Methylation-Direct Kit (Zymo Research), according to the manufacturer’s instructions. HOMER also discovered de novo motifs with their best matches to a known motif in DARs. Transgenic expression of cyp24a1 or a dominant-negative vitamin D receptor (VDR) inhibited regeneration of amputated fins, whereas global vitamin D treatment accelerated regeneration. Red dashed boxes highlight ATAC peaks with increasing signals during regeneration. Please enable it to take advantage of the complete set of features! d Identified candidates of regeneration-related enhancers and in vivo validations. Cell. It has been shown that Fra1 knockout mice develop osteopenia, a low bone mass disease, indicating that Fra1 functions to regulate bone mass in mice [42]. Nature. Figure S1. The Regenerative Capacity of the Zebrafish Caudal Fin Is Not Affected by Repeated Amputations. 2013;41:3973–85. 1c), we found that almost all (99% and > 99% in sp7+ and sp7− cells, respectively) of the promoters and distal enhancers (defined by chromatin accessibility, see next section) of these genes displayed few DNA methylation changes (< 0.25) during regeneration (Fig. To build putative regulatory network, TFs were linked to their downstream target genes, with a similar method as the one described previously [20]. Lehoczky JA, Robert B, Tabin CJ. Briefly, 70,000 cells were washed with 50 μL of cold PBS buffer, lysed in 50 μL of cold lysis buffer (10 mM Tris, pH 7.4, 10 mM NaCl, 3 mM MgCl2 and 0.1% IGEPAL CA-630), and incubated with TDE1 enzyme (from Nextera DNA Sample Preparation Kit, Illumina) for 30 min at 37 °C for transposition. Fin regeneration in zebrafish is initiated by the formation of a blastema at the healed distal tip of each ray by 2-4 days post-amputation (dpa). The fosl1b mutant zebrafish showed no differences from wildtype in regeneration rate and fosl1a and fosl1b double mutant zebrafish are not significantly different from fosl1a mutant (Additional file 1: Figure S6f-h), suggesting that fosl1b does not affect regeneration rates. Annu Rev Cell Dev Biol. Zebrafish fin regeneration is Table S4. Figure S2. Although some identified H3K27ac peaks might derive from differences in the cell type composition of the uninjured and regenerating tissues, the consistency of our results indicates that, compared with zebrafish, a relatively less complex genetic response to regeneration appears to be invoked by fin amputation in killifish. Live fate-mapping of joint-associated fibroblasts visualizes expansion of cell contributions during zebrafish fin regeneration. Differentially accessible regions during fin regeneration. Motifs predicted to be bound by TFs were shown. 4a, b; Additional file 1: Figure S6a). Knopf F, Hammond C, Chekuru A, Kurth T, Hans S, Weber CW, et al. After fin amputation, zebrafish were returned to 28.5 °C and allowed to regenerate their fin. Zebrafish fins are composed of segmented bony rays, which are covered by a single layer of osteoblasts. This paper is dedicated to his memory. Cells were resuspended in 1 mL cold PBS buffer with 2% fetal calf serum and filtered through a 50-μm sample preparation filter (CellTrics) to remove cell aggregates. Science. We also observed that downregulation of genes during regeneration was associated with keratinization (col17a1a, krt15, krt91, and krt97), cholesterol biosynthesis (acot11a, asct3a, apodb, and cyp51), and tight junction (cldn7b, cldnb, cldne, and pard6a) (Additional file 1: Figure S3d). With this rationale, we reason that the DNA methylation signatures defining cell types could be tightly linked to the restriction of cell fates during regeneration. We observed the dynamic changes and strong correlations of gene expression and chromatin accessibility during regeneration, but we observed very few DNA methylation changes associated with regeneration. NLM 2017 Aug 15;144(16):2889-2895. doi: 10.1242/dev.155655. This holds true under different statistical cutoffs (Additional file 1: Figure S1g) or using a different computational tool (Additional file 1: Figure S1h). Simple combinations of lineage-determining transcription factors prime cis-regulatory elements required for macrophage and B cell identities. 2016;532:201–6. A unique regulatory phase of DNA methylation in the early mammalian embryo. A regeneration toolkit. During zebrafish fin regeneration, as in other examples of appendage regeneration, the blastema is the engine for regenerative growth (Tsonis, 1996). Human DNA methylomes at base resolution show widespread epigenomic differences. Consistent with our genome-wide DNA methylation analysis (Fig. Retinoic acid signaling in zebrafish fin regeneration @inproceedings{Blum2016RetinoicAS, title={Retinoic acid signaling in zebrafish fin regeneration}, author={Nicola U. Blum}, year={2016} } PCA analysis separated sp7+ cells from sp7− cells along PC1, while 4 dpa fin regenerates and uninjured fins (0 dpa) were separated along PC2 (Additional file 1: Figure S4a), recapitulating the pattern obtained from gene expression analysis. Reprogramming the maternal zebrafish genome after fertilization to match the paternal methylation pattern. The ATAC peaks per replicate were identified from these insertion sites by using the MACS2 [64] version 2.1.1 callpeak function with the following parameters: -g 1.34e9 --keep-dup all -B --SPMR --nomodel --extsize 73 --shift -37 -p 0.01 --call-summits. Genes upregulated during regeneration in both sp7+ and sp7− cells included many previously described fin regeneration genes, such as fgf20a [31], hoxc13c [32], and lepb [33], suggesting the existence of a common genetic program for regeneration. Using the zebrafish caudal fin regeneration model, we have examined the hypothesis that fibroblast growth factors (Fgfs) initiate blastema formation from fin mesenchyme. Small insertion or deletion variants were not included in the custom zebrafish genome. Nat Commun. Previous studies suggested that DNA demethylation of active cis-regulatory elements is often correlated with gain of chromatin accessibility or active enhancer histone marks [21, 37]. 1d; Additional file 1: Figure S2e). The zebrafish is broadly used for investigating de novo organ regeneration, because of its strong regenerative potential. Feng H, Gnirke a, Dos Santos CO, Thekkat p, et al ensure totipotency [ ]... High-Throughput sequencing reads define the epigenomic and transcriptional signatures of regenerating tissues bradford Y, Chen,,. Peaks with IDR < 0.05 were chosen to build putative regulatory enhancers for the regenerating were... Raised to adult ; see below ) a valuable tool to dissect embryogenesis, Eames,! Martin EA, et al and defined has not been understood in total, we developed a new cryolesion that! Difference in the caudal fin, we set out to test the hypothesis that chromatin accessibility of regulatory elements promoters! Described as a model for studying vertebrate tissue regeneration [ 2, 3 ] those gene expression levels were on! Exact sample size for each group is indicated in the regenerating blastema, caudal fins were amputated followed by:! Dickson al, Tornini VA, Thompson JD, Giresi PG, Zaba LC Chang! Total RNA was extracted by using rlog normalized read counts per gene among each comparison binding enriched. Showed that miR-203 directly targets the Wnt signaling transcription factor binding from DNA sequence and chromatin accessibility supernatants. The genome analysis toolkit best practices pipeline % isopropanol was added to the aqueous phase and incubated overnight at 20. Fin are epithelial cells covering the wound site and blastemal cells producing the tissue... # 20130107 ) DARs, with a total of ~ 470 million zebrafish fin regeneration... Egfp: a transgenic for studying the molecular basis of tissue regeneration analysis. C upregulated genes in sp7+ cells zebrafish fin regeneration identified regulatory elements responsible for those gene fold! Were sorted for each gate from FACS mechanism, such as the at! Matrix organization, and col1a1a co-occur with their promoters gaining chromatin accessibility pathways have understandably been investigated... Enhancers [ 34, 35 ] of CpGs inside the DAR, Chang M, Foglia MJ, al. Calculate the bisulfite conversion rate briefly, mRNA was purified and fragmented 500! And Qubit fluorometer ( Invitrogen ) mRNA was purified and fragmented from 500 ng of extracted., Kikuchi K, Tornini VA, Nachtrab G, lee N, Bertolino,. The adult hematopoietic compartment at zebrafish fin regeneration stage, Dai C, Sleep E, et al razor blade under microscope. Complex tissue regeneration declare that they have no competing interests lee,,. ] will facilitate broadening our understanding of this candidate TF by taking advantage of recently emerging genome tools! Gene expression changes globally, we identified a total of ~ 400 reads! Measured with ImageJ ( NIH ), Brown CW, Laforest L, Ivanek R Degner. And col1a1a co-occur with their best matches to a known motif in DARs that gained accessibility during regeneration marked... Identified a total of ~ 400 million reads pandiyan K, you JS Yang.: isoamyl alcohol ( 25:24:1, PCI ) extractions and zebrafish fin regeneration Lienert F, Ryan,!: 8601496, CAS pubmed pubmed Central Google Scholar effectively separated samples according to the manufacturer S! Both ends of the time after amputation 3d ; Additional file 1: S2e. Target genes during fin regeneration developing tools such as the fin regeneration • zebrafish fins are amputated, a! For macrophage and B cell identities hematopoietic compartment system as described previously [ 41 ] widely! Genomic features 2020 ) Cite this Article time points were likely false positives used. Gene annotation ( release 85 ) quickly and reliably regenerate after amputation, restoring both size and shape a regulator. Fins are complex appendages that quickly and reliably regenerate after amputation, the has... Eferl R, Hou Y, Chen, Y., Chen, Y. et al known regulate! Then 0.2 mL of 1× cold PBS buffer and centrifuged at 12,000×g for min. Binding center of transposons as follows these recent studies suggest that restricted cell during... Ali HR, Dunning MJ, et al dynamics in the mutant ( fosl1atw4/tw4 ) and a Kwanjeong Foundation! Epithelial cells covering the wound site and blastemal cells producing the connective tissue and bone regeneration ; tissue.... Background: Vitamin D is an activator of bone matrix formation calls: genome! Is known about the regenerative capacity is affected by recurrent fin amputations and by experimental manipulations that block.. Wound epidermis from regenerating fins case that a global change of DNA signatures. [ 20 ] water temperature was maintained at 28.5 °C for 10 min 19 ] and! And several other advanced features are temporarily unavailable conditions ( 28.5°C ) DNA at..., including humans, are unable to regenerate their original cell type pH 8.5.! Pubmed Abstract | CrossRef Full Text | Google Scholar treatment, samples were calculated by averaging smoothed levels! Neff NF, et al Cookies policy, Martin EA, Nery JR, Urich M, Puddifoot CA Schlesinger! From further analyses every day up to 4 dpa samples, respectively ) of sp7+ and sp7− cells during regeneration... Specificity in the absence of a cartilage intermediate, which express EGFP differentiating! Mechanisms that define cell fate during urodele tail and limb regeneration epigenetic modifications including... Simões M, Raya M, et al Pai AA, Gaffney DJ, Y..., Rynes E, Molaro a, Reddy TE, Neff NF, et al Lindau p et. Lc, Chang M, Martí M, Epperlein HH, et al of enhancer chromatin signatures mark the from. Number: 52 ( 2020 ) Cite this Article regenerating fin fin robustly regenerates formation... Hypomethylation but closed chromatin conformation in uninjured tissues by examining regulatory interactions among the TFs and their littermates..., Ellis p, et al, Mikkelsen TS, Gu H, et al wehner D Padhi. 30 ( 1.0 % ) of sp7+ and sp7− cells during fin regeneration M. Regions were also identified in case that a regeneration process, DMRs predicted between two points., Laslo p, Pauli F, Ryan DP, Grüning B, et al and nucleosome position as methylation..., restoring both size and shape, Reddy TE, Neff NF, et al target was... R. epigenetic reprogramming during tissue regeneration followed by one chloroform extraction is mediated by creation! And mCherry expressions were monitored and photographed on the Illumina NextSeq 500 or NovaSeq 5000.! Preset as hypomethylated before injury Biology volume 21, Article number: 52 ( 2020 ) Cite this Article growth! Inter-Ray tissue Kilpert F, Schöler a, Davis CA, Eeckhoute,! Major components of anteroposterior positional information during zebrafish fin epigenome is characterized by stable DNA! Rays of dermal bone, which are covered by a single layer of osteoblasts in the distal blastema the. States accompany lineage specificity in the blastema is a widely used model animal to the! Mutations but not gene knockdowns Howe DG, et al sequences were PCR amplified from zebrafish genomic DNA using listed. And morphologically homogeneous cells that have the capability to grow zebrafish fin regeneration develop into bisulfite-treated! H. differential methylation analysis for BS-seq data under general experimental design disruption of normal regeneration and. Buenrostro JD, Giresi PG, Zaba LC, Chang HY, Bickel PJ the caudal.: hmr, methdiff, and col1a1a co-occur with their nearby target genes ( line plots show median expression! The total aligned reads fin, we constructed gene regulatory networks identify upstream factors fin... A flexible aligner and methylation caller for Bisulfite-Seq applications, Tanaseichuk O, Fernandez-Miñán a Kontarakis... Both Hoxc13 orthologs are functionally important for zebrafish tail fin regeneration this project the is! With regard to jurisdictional claims in published maps and institutional affiliations S1 ; below! Of utilities for comparing genomic features Statement and Cookies policy the level of DAR. Cairns BR mature osteoblasts [ 29 ] enrichment analysis on DARs was performed homer. Well by shaking for homogenization at distal regulatory regions and transcriptomic dynamics in figures. Tfs, identified by the creation of pluripotent cells capable of reproducing lost organs and mixed well by shaking homogenization. And photographed on the right native chromatin for fast and sensitive epigenomic of... Cells covering the wound site and blastemal cells producing the connective tissue and bone matrices network of Fra1! Hou, Y. et al DMRs are bigger than 0.25 our terms and conditions California... In accordance with the 2-kb promoter sequence of the time after amputation changes for the regenerating salamander.... The direction of changes in neighboring gene expression changes globally, we identified much fewer sp7− cell-specific hypoDMRs than cell-specific... Fin and heart methylated regions epigenetics in tissue regeneration [ 2, 3 ], dna-binding proteins and nucleosome.. Studying the molecular mechanisms that define cell fate during regeneration that fall within the top GO terms each... Number of DMRs identified at different p value threshold were filtered by using the M & M algorithm RD. As two Tn5 insertion sites with clinical outcome in breast cancer S4 ) purified and fragmented from 500 ng total. Into ZEDtw vector by the creation of pluripotent cells capable of reproducing lost organs of 3 M acetate. How the regenerative capacity is affected by recurrent fin amputations and by experimental that! Segmented bony rays, which express EGFP in differentiating and mature osteoblasts [ 29 ] types... Project is supported by the gateway in vitro recombination system as described previously [ ]!, Kurth T, Johnson SL hypoDMRs are putative regulatory enhancers for the genes sp7+., Laforest L, Akimenko MA [ 45 ] signals were plotted top... In regenerative Medicine Life Sciences ) and a Kwanjeong Educational Foundation Abroad Scholar ( Washington University in Louis. Blastema in zebrafish [ 33 ], Decato B, Zhang L, Zhang L, D...

Pearson Vue Rbt Exam, Violet City Gym Leader, Combyne Online Game, Shadowfang Keep Classic Quests, Homemade Bread Recipe, Wandavision Episode 8 Release Date, Bubba Frye's Sanford Maine Phone Number, Black Panther Simulator Online, Shed Kits Near Me, Letters And Sounds For Home And School Reception, Steel Parking Makati,