Ecol. Together, while the Hox A, B and D clusters exhibit the canonical, conservative nature even in elongated elasmobranch genomes, their Hox C cluster underwent remarkable lineage-specific, sequence-level modifications. Although the entire Hox C cluster was reportedly missing from elasmobranch genomes19,20, we identified putative Hox C genes in the genome and transcript sequences of the analysed shark species (Fig. Samples of the cloudy catshark Scyliorhinus torazame were supplied by captive breeding at the Aquarium Facility of RIKEN Center for Developmental Biology and Atmosphere and Ocean Research Institute of University of Tokyo. Biol. 4a). Kuraku, S., Usuda, R. & Kuratani, S. Comprehensive survey of carapacial ridge-specific genes in turtle implies co-option of some regulatory genes in carapace evolution. Nutr. Genome Biol. The assembly step employed paired-end reads and single reads whose pairs had been removed, and the scaffolding step employed paired-end and mate-pair reads. Overall, these findings indicate that despite the variable genome sizes and repetitive element compositions, elasmobranch genomes have undergone less modification in noncoding regions involved in gene regulation since the jawed vertebrate ancestor than is inferred by their evolutionary distance. 1e). ChIP assays, as well as ChIP-seq and downstream data analysis, were performed as previously described15. Davies, W. L. et al. Internet Explorer). Biol. Endocrinol. Neurobiol. Huggel, H. Experimentelle untersuchungen ueber die automatie, temperaturabhaengigkeit und arbeit des embryonalen fischherzens, unter besonderer beruecksichtigung der salmoniden und scylliorhiniden. Not satisfied with your Sharks? 33, 290295 (2015). Bioinformatics 27, 10171018 (2011). Mol. We also queried the lncRNA candidates against the Augustus-predicted coding genes of the bamboo shark genome and removed sequences if the best hits were aligned with the forward strands of the predicted coding genes. Memories Of Vortex (El Payo Remix) 30. The developmental staging was performed according to existing literature for small-spotted catshark S. canicula56 and our original table for C. punctatum8. Previously, two short wavelength-sensitive opsin genes, SWS1 and SWS2, were found to be missing in the C. milii genome42. & Furness, J. 3fi and Supplementary Fig. Yokoyama, S., Tada, T., Zhang, H. & Britt, L. Elucidation of phenotypic adaptations: molecular analyses of dim-light vision proteins in vertebrates. Kim, D., Langmead, B. 69, 122130 (2017). Narendra, V. et al. 4a and Supplementary Note 17). & Baker, C. V. A molecular analysis of neurogenic placode and cranial sensory ganglion development in the shark, Scyliorhinus canicula. The numbers of PCR cycles and conditions of size selection for individual libraries are included in Supplementary Table 1. Elasmobranchs have scarce repertories of opsin and olfactory receptor genes, possibly associated with their unique niche. To infer individual gene family trees, amino acids sequences were retrieved from aLeaves90 incorporating Ensembl release 84. For elasmobranchs, however, no reliable genome-wide sequence resource allowing extensive molecular analyses has been established to date, in spite of some attempts3,4. SkateBase, an elasmobranch genome project and collection of molecular resources for chondrichthyan fishes. 34, 27732791 (2017). To build homologue hints for the cloudy catshark, we used a set of 117,246 NCBI RefSeq protein sequences downloaded on 23 November 2015, including known human proteins (39,582 sequences), chicken (6,189 sequences) and amniote vertebrates (44,675 sequences), as well as C. milii (NCBI Genome version 6.1.3, 26,800 sequences). We removed sequences of antisense RNAs that overlapped open reading frames in the lncRNA database and repetitive sequences that were masked with RepeatMasker v4.0.5, as described above. Well be checking link in bio for slides, DONT LOOK AT THE TEMPLATE link in bio for sharks , Why is nobody talking about this? Correspondence to & Stadler, P. F. Exclusion of repetitive DNA elements from gnathostome Hox clusters. Kuraku, S., Qiu, H. & Meyer, A. Horizontal transfers of Tc1 elements between teleost fishes and their vertebrate parasites, lampreys. Their whole genomes, measured to be approximately 4.7 and 6.7Gbp, respectively, were sequenced de novo to obtain assemblies including megabase-long scaffolds (Supplementary Note 1.1). Blood of bamboo shark and catshark was sampled from the heart using a 1ml syringe with a 21G needle and immediately diluted 1:10 in shark saline solution containing 2mM EDTA. The cell suspension was filtered through a 40m cell strainer (BD Bioscience) to remove cell clumps and debris. 11, 265275 (2015). #fyp #fyp #foryoupage #cloudysharks #scammer #cloudysharkscam #foryou #foryoupage 150 Likes, 78 Comments. Memories Of Vortex (El Payo Remix (Instrumental)) 4. First, peptide sequences of the retrieved orthologues were aligned with MAFFT v7.299b86 with the option -linsi. Designed with a concave sole that protects your toes from kicking off table legs or door sills. Following the BLASTN searches, the best hits whose bit scores exceeded 60 were counted. Genome Res. Grab your own Shark Slide today. 10, R25 (2009). Gen. Comp. PubMed Bioinformatics 25, 19721973 (2009). 4b,c). Mate-pair libraries were prepared using a Nextera Mate Pair Sample Prep Kit (Illumina), employing our customized iMate protocol59 (http://www.clst.riken.jp/phylo/imate.html). 25, 33893402 (1997). . 5a) expressed in a temporally collinear manner (Fig. CAS a, Number of the retained CNEs identified by the humanchicken genome comparison. Science 353, 702704 (2016). Dev. Our comparisons of coding sequences detected a higher similarity in nucleotide and amino acid compositions of sharks to tetrapods and coelacanth than to actinopterygian fishes (Supplementary Fig. Our shoes are so comfy, stylish, and versatile. Sci. In situ hybridization using whole-mount embryos and paraffin-embedded sections of 8m thickness was performed as previously reported83,84. These local genomic characteristics may have facilitated the secondary loss of the coding genes embedded in the divergent genomic regions. This coding/noncoding association was detected for the divergent nature of FoxG2, while the level of sequence conservation of FoxG3 was intermediate (Fig. Unique evolution of neurohypophysial hormones in cartilaginous fishes: possible implications for urea-based osmoregulation. Our spectroscopic analysis of the RHO pigments revealed blueshifted spectra for not only the cloudy catshark (max, 484nm) but also the whale shark (478nm) that occasionally migrates down to the bathypelagic zone (~2,000m) besides daytime surface feeding habits50 (Fig. Sci. PLoS ONE 9, e92751 (2014). Mol. Langmead, B., Trapnell, C., Pop, M. & Salzberg, S. L. Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. When multiple motifs were identified within a peak region, only the motif with the lowest p value was adopted for downstream analyses. RNA-seq data used for exon hint construction is indicated in Supplementary Table 6. Cell Rep. 19, 17231738 (2017). Resultant genomic scaffold sequences were screened for contaminating organismal sequences, PhiX sequences loaded as a control, mitochondrial DNA sequences, and those shorter than 500bp, as performed previously28. Replying to @geo.rge_came.home Who wears there slides with thier dogs out? Tatsumi, N. et al. More than a decade ago, the elephant fish, Callorhinchus milii, a member of the Holocephali that comprises approximately 50 species, was chosen for whole-genome sequencing because of its small genome size1. Google Scholar. 16, 40 (2018). 2, 288298 (2018). Rep. 7, 4957 (2017). ISSN 2397-334X (online). Article The bamboo shark transcriptome data demonstrated that most of the clustered Pcdh genes consist of both variable and constant exons and exhibit a biased expression pattern towards neural tissues, as previously shown for mammalian and teleost counterparts (Supplementary Fig. B. Ghrelin and motilin receptors as drug targets for gastrointestinal disorders. Phyloinformatics Unit, RIKEN Center for Life Science Technologies (CLST), Kobe, Japan, Yuichiro Hara,Kazuaki Yamaguchi,Koh Onimaru,Mitsutaka Kadota,Sean D. Keeley,Kaori Tatsumi,Kaori Tanaka,Fumio Motone,Yuka Kageyama,Osamu Nishimura,Reiko Nakagawa,Chiharu Tanegashima&Shigehiro Kuraku, Laboratory for Phyloinformatics, RIKEN Center for Biosystems Dynamics Research (BDR), Kobe, Japan, Yuichiro Hara,Kazuaki Yamaguchi,Koh Onimaru,Mitsutaka Kadota,Kaori Tatsumi,Osamu Nishimura,Reiko Nakagawa,Chiharu Tanegashima&Shigehiro Kuraku, Department of Biology and Geosciences, Graduate School of Science, Osaka City University, Osaka, Japan, The OCU Advanced Research Institute for Natural Science and Technology (OCARINA), Osaka City University, Osaka, Japan, Graduate School of Science and Technology, Kwansei Gakuin University, Sanda, Japan, Okinawa Churashima Research Center, Okinawa Churashima Foundation, Okinawa, Japan, Okinawa Churaumi Aquarium, Okinawa, Japan, Ryo Nozu,Rui Matsumoto,Kiyomi Murakumo&Keiichi Sato, Evolutionary Morphology Laboratory, RIKEN Cluster for Pioneering Research (CPR), Kobe, Japan, Laboratory for Evolutionary Morphology, RIKEN Center for Biosystems Dynamics Research (BDR), Kobe, Japan, Laboratory of Physiology, Atmosphere and Ocean Research Institute, University of Tokyo, Kashiwa, Japan, You can also search for this author in After centrifugation at 500g for 5 min, blood cells were washed once in shark saline solution containing EDTA and counted. 158K Followers, 24 Following, 28 Posts - See Instagram photos and videos from CloudySharks - The Orginal Shark Slides (@cloudysharks) The authors declare no competing interests. Frith, M. C. & Kawaguchi, R. Split-alignment of genomes finds orthologies more accurately. Philos. Bioinformatics 30, 566568 (2014). DNA staining was performed by adding 1ml of PI/RNase staining buffer (BD Bioscience). Smit, A. F. A. Total RNAs were extracted with Trizol reagent (Thermo Fisher Scientific). Mayor, C. et al. Rep. 6, 30580 (2016). Shark genomes provide insights into elasmobranch evolution and the origin of vertebrates, https://doi.org/10.1038/s41559-018-0673-5. The peaks overlapping between replicates were identified by bedtools v2.19.173 and designated as consensus peaks. Fried, C., Prohaska, S. J. Biotechnol. Link in bio , Which color is your favorite? Nishimura, O., Hara, Y. Biol. In the meantime, to ensure continued support, we are displaying the site without styles Closed pink boxes, presence of intact genes; grey boxes, pseudogenization indicated by a stop codon inside the homeobox (see Supplementary Fig. Biol. Durable and light Relief foot and joint ache Weatherproof Flexible for indoor/outdoor $38.99 $33.99 You save 15% Size guide color : heather gray Replying to @xtrme.dk2 W or L? 5a). Our thorough genome annotation revealed Hox C genes previously hypothesized to have been lost, as well as distinct gene repertories of opsins and olfactory receptors that would be associated with adaptation to unique underwater niches. Completeness of the genome assemblies was assessed with (1) CEGMA v2.564, (2) BUSCO v2.0.165 and (3) a manual curation-based census of Wnt genes (Supplementary Fig. We have a first-class R&D team, constantly innovating our shark series products, whether it is summer or winter, you can find the style you like, including other animal slippers, everything! These new designs will only be released for a limited time only, so hurry and get yours hands on Cloudy Shark Slides have been absolutely going viral worldwide on Tiktok. All authors contributed to the final manuscript editing. 62, 611615 (2013). 8). Sanger, G. J. B Mol. 27, R565R572 (2017). Bootstrap values were 100 for all nodes. j, Repetitiveness of the whole genomes and the Hox-containing genomic regions (see Methods and Supplementary Note 16.4 for details). Mate-pair reads were processed by NextClip v1.160 with default parameters. 15, R82R83 (2005). The Original CloudySharks $29.99 $49.95. About Press Copyright Contact us Creators Advertise Developers Terms Privacy Policy & Safety How YouTube works Test new features NFL Sunday Ticket Press Copyright . Biol. ------------------ These sandals are sure to become your go-to for an idyllic day at the shore or a relaxed amble about town. Article 7a for a full heatmap). Niimura, Y. Molecular phylogenetic trees were inferred by RAxML with the -m PROTCATWAG -f a -# 1000 options unless stated otherwise. Google Scholar. Since then, molecular comparative studies on vertebrates have largely relied on the C. milii genomic sequences as representative of cartilaginous fishes2, but the low fecundity and accessibility of live specimens have been a limitation. Gesta, S., Tseng, Y. H. & Kahn, C. R. Developmental origin of fat: tracking obesity to its source. Our study has provided an unprecedented set of genomic, transcriptomic and epigenomic data from three elasmobranch species, with the bamboo shark genome assembly achieving the highest continuity. Bioinformatics 33, 36353637 (2017). These included prolactin (PRL1), orexin, kisspeptin, spexin, motilin and prolactin receptor implicated in fertility, appetite, digestion and sleep in mammals29,30,31, as well as osmolarity and gastrointestinal control in teleost fishes (Supplementary Note 18). i, Expression timings of the Hox11 genes in whole catshark embryos. 19, 95105 (2018). Trans. Scale bars, 500m. Two left panels: the numbers of non-synonymous and synonymous substitutions per site (KA and KS, respectively) of bamboo shark and catshark orthologue pairs that were calculated with codeml in PAML v4.9c88. and S.Kuraku drafted the manuscript. & Hubley, R. RepeatModeler Open-1.0 (20082010); http://www.repeatmasker.org, Smit, A. F. A., Hubley, R. & Green, P. RepeatMasker Open-4.0 (20132015); http://www.repeatmasker.org. Comment below! 1d). CloudySharks Follow 22 Following 110K Followers 1.9M Likes The Original Shark Slippers! CloudySharks Cozy Editions Out Now myhalobae.com/products/shark-cloud-slides?variant=42216719286440 Videos Liked Playlists Cozy Edition Drop 10 posts Videos 3423 Cloudy shark slides in bio! placeholder. The gap closure step employed all of the single, paired-end and mate-pair reads. Zool. C. milii is often referred to as elephant shark (or ghost shark), but true sharks belong to the subclass Elasmobranchii that comprises approximately 1,200 species. We focused on the brownbanded bamboo shark Chiloscyllium punctatum, for which we recently tabled embryonic stages8, and the cloudy catshark Scyliorhinus torazame. 7, 317 (2005). Quality control of DNase I-treated RNA was performed with Bioanalyzer 2100 (Agilent Technologies). We also used human GM12878 cells as a reference, which were cultured in RPMI-1640 media (Thermo Fisher Scientific) supplemented with 15% FBS, 2mM l-glutamine, and 1 antibiotic-antimycotic solution (Gibco) at 37C with 5% CO2. 6 and Supplementary Fig. Replying to @abbi_pogue Whos ready for the summer time? Fluorescence intensities were measured with the excitation at 488nm and the bandpass filter of 575/26nm on a FACSCanto II cell sorter (BD Bioscience). c, Expression profiles of the putative bamboo shark homologues of human lncRNAs expressed in at least one analysed tissue. 283, 71857195 (2008). This identified long interspersed nuclear elements as the most abundant class of repetitive elements, exceeding the proportions of long terminal repeats and those unclassified into any existing repetitive element class, both of which were particularly expanded in the catshark (Fig. Sadaie, M., Shinmyozu, K. & Nakayama, J. Evol. 24, 15861591 (2007). Mouse embryonic fibroblast cells, used as a reference, were prepared from E14.5 embryos and cultured in DMEM media supplemented with 10% FBS, at 37C with 5% CO2. Nat Ecol Evol 2, 17611771 (2018). Data 5, 180197 (2018). Wherever your. Z. Vgl Physiol. VII+VIII, acoustico-facial ganglionic complex; X, vagal ganglion; tel, telencephalon; ret, retina; ot, otic vesicle; GC4, GC-content at four-fold degenerate sites. Dashed blank boxes indicate the absences of orthologues in the currently available genome assemblies. 6 and Supplementary Note 19). Nat. Remarkably, none of the identified, putative shark Hox C genes comprised such a compact cluster as in other jawed vertebrates, spanning within a 100-kbp-long genomic region13for example, catshark Hoxc11 was flanked by a 50-kbp-long stretch containing no other Hox gene (Supplementary Fig. Likewise, for the gene prediction of the bamboo shark, we incorporated the predicted whale shark peptide sequences into the sequence set used in gene prediction for the catshark. Zool. Grant, C. E., Bailey, T. L. & Noble, W. S. FIMO: scanning for occurrences of a given motif. Overall, we identified the orthologues of almost all hormones and their receptors involved in the hypothalamopituitary and gastrointestinal systems documented mainly in mammals (Fig. in The Elasmobranch Husbandary Manual II: Recent Advances in the Care of Sharks, Rays and their Relatives (eds Smith, M. et al.) PubMed Central Replying to @jadalynne25 Why dont you have a pair of our shark slides yet? See Supplementary Note 19.1, for details of the habitat depths and the absorption spectra obtained by our spectroscopic analysis. Cloudyshark Studios is a rapidly growing social mobile game developer and publisher based in Austin, Texas. CAS rated 5-star by 362+ reviews. Lightweight anti-slip sole due to EVA material, which provides long-lasting comfort and flexibility for tired toes and heels, also it absorbs liquid easily, thus drying quickly, has a cooling effect and it's quite sturdy - invaluable qualities on hot summer days. 9, 678685 (2008). Hara, Y., Yamaguchi, K., Onimaru, K. et al. If you love free stuff We're serious ;). Memories Of Vortex. Nature 505, 174179 (2014). By using the program BLASTN 2.5.0+with the dc-megablast mode, we queried the refined set of lncRNAs against the genome assemblies of other vertebrates used above in CNE detection. Kherdjemil, Y. et al. 6). These were screened for transcript evidence in bamboo shark RNA-seq data and absence of homology to coding sequences. Protoc. Evol. Hundred Grand to the Man. MALAT-1, a novel noncoding RNA, and thymosin beta4 predict metastasis and survival in early-stage non-small cell lung cancer. Parra, G., Bradnam, K. & Korf, I. CEGMA: a pipeline to accurately annotate core genes in eukaryotic genomes. The assessments were executed on the gVolante web server67 (Supplementary Note 2) using a script released by us previously66. 30, 772780 (2013). Wyffels, J. et al. a, Structure of the bamboo shark Hox clusters. Bernard, V., Young, J., Chanson, P. & Binart, N. New insights in prolactin: pathological implications. 3d). A., Carlisle, H. R. & Dahn, R. D. A natural deletion of the HoxC cluster in elasmobranch fishes. 9, R137 (2008). Construction of gene models on the cloudy catshark, whale shark and bamboo shark genomes was performed in this order, following the procedure previously reported15 (Supplementary Note 5). Biol. 16, R484R489 (2006). Orthologue groups containing fewer than 50 aligned codons or a stop codon were discarded. After size selection, the quantification of the prepared libraries was performed using a KAPA Library Quantification Kit (KAPA Biosystems). Y.H., K.Y., S.D.K., M.Kadota, M.Koyanagi, S.H., K.S., K.O. CloudySharks are a fun, colorful and comfy way to wear your shoes. Elephant shark sequence reveals unique insights into the evolutionary history of vertebrate genes: a comparative analysis of the protocadherin cluster. Methods Mol. Bioinformatics 16, 10461047 (2000). This inference marks leptin, a key metabolic and neuroendocrine regulator in mammals34, and the signalling cascade through its receptor (LepR) as an invention in the jawed vertebrate lineage (Supplementary Note 18.10). Opin. See Supplementary Notes 13 and 14 for details. Bioinformatics 33, i75i82 (2017). Multiple sequence alignment was performed with MAFFT with the option -linsi. Designed for all ages, men and women alike, the CloudyShark is an easy today. The purple dots indicate the branches b1b4 in d. f, Genome size and intron length. Nat. Ebert, D. A., Fowler, S. & Compagno, L. Sharks of the World: A Fully Illustrated Guide (Wild Nature Press, Plymouth, 2013). b, Expression profiles of hormone genes in an embryo and adult tissues of the catshark (see Supplementary Fig. Cartilaginous fishes (Chondrichthyes) are divided into two subclasses, elasmobranchs (Elasmobranchii, including sharks, rays and skates) and chimaeras (Holocephali), and their common ancestor diverged from the rest of jawed vertebrates around 450 million years ago. For the selected orthologue groups, KS were computed with codeml in the PAML v4.9c88. the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in J. Biol. 6cp). Visualization of sequence similarity between species employed VISTA75 using a global pairwise alignment program Shuffle-LAGAN76. BMC Genomics 16, 977 (2015). 5gi) are expressed in the posterior regions concomitantly with their sister paralogues in the Hox A, B and D clusters. Curr. h, Breakdown of the whole-genome assemblies into different segments. Together, the more prone a FoxG paralogue is to secondary loss, the more restricted is its expression domain in shark embryos. These alignments were passed to StringTie v1.3.095 to generate an additional set of tissue-specific gene models. Rev. While the determinant of loss or retention of gene duplicates is often imputed to their functions27, the effect of intrinsic genomic characteristics, independent of gene functions, has also been proposed as a cause28. It's not clear how the two escaped the shark and returned to shore. Genome Biol. This screening resulted in the identification of 38 transcript contigs with variable degrees of spatial expression biases (Fig. Biol. Without rain falling into the ocean and diluting it with freshwater, the salinity (salt content) of ocean water close to shore becomes more concentrated, or saltier than usual. Genome Res. FoxG3 was expressed in an anterodorsal part of the retina, in addition to the FoxG2-positive domains (Fig. Our study showed that elasmobranchs contain slightly higher numbers of Pcdh genes in markedly longer clusters than osteichthyans (Supplementary Fig. Our products will fuel diverse life science studies on sharks and evolutionary investigation about early vertebrates. Sharks Like It Saltier With Low Rainfall. Nov 2022. Bioinformatics 18, 4.3.14.3.28 (2007). NY. Pan, W. W. & Myers, M. G. Jr. Leptin and the maintenance of elevated body weight. Play. Evolution of Hox gene clusters in gnathostomes: insights from a survey of a shark (Scyliorhinus canicula) transcriptome. Step up your summer style in the CloudySharkscomfy, stylish and breathable enough for the beach. FBS was added to stop digestion with trypsin. & Heithaus, M. R.) Ch. They were sequenced on a HiSeq 1500 (Illumina), operated by HiSeq Control Software v2.0.12.0 using a HiSeq SR Rapid Cluster Kit v2 (Illumina) and HiSeq Rapid SBS Kit v2 (Illumina), and MiSeq operated by MiSeq Control Software v2.3.0.3 using MiSeq Reagent Kit v3 (600 Cycles) (Illumina). For both CEGMA and BUSCO, we employed not only the reference gene sets provided inherently with these program pipelines, but also the core vertebrate genes introduced particularly for vertebrates, especially species in isolated lineages such as elasmobranchs66. d, Temporal expression patterns of Hox A genes in whole catshark embryos. We used cells prepared from the liver and blood of a 32-cm-long juvenile female bamboo shark, the blood of a 5-m-long live female whale shark reared in Osaka Aquarium Kaiyukan, and the liver and blood of a 16-cm-long juvenile male cloudy catshark (see above for the detail of sampling). 5m for scaffold IDs for Hox C). Capella-Gutierrez, S., Silla-Martinez, J. M. & Gabaldon, T. trimAl: a tool for automated alignment trimming in large-scale phylogenetic analyses. Stamatakis, A. RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. contracts here. Zhang, X., Hamblin, M. H. & Yin, K. J. Measurements were carried out with three technical replicates per sample, and the acquired values were averaged before DNA content calculations (Supplementary Table 4). Train, C. M., Glover, N. M., Gonnet, G. H., Altenhoff, A. M. & Dessimoz, C. Orthologous Matrix (OMA) algorithm 2.0: more robust to asymmetric evolutionary rates and more scalable hierarchical orthologous group inference. 4b and Supplementary Table 15). Evol. Catshark and bamboo shark embryos were fixed with 4% PFA/PBS, dehydrated with methanol series and stored in 100% methanol at 30C until use. Nat. We also show the early establishment of the genetic machinery governing mammalian homoeostasis and reproduction at the jawed vertebrate ancestor. and 17K07426 to S.Kuraku. Our mission is to create innovative games that are emotionally engaging, accessible, and . The number of best hits that were longer than 100bp was counted for each of the two search modes. In addition, although typical jawed vertebrate Hox clusters are almost free from repetitive elements21 (at most 2.9% in length for elasmobranch Hox A, B and D clusters; Fig. Oncogene 22, 80318041 (2003). Our gratitude extends to K. Muguruma, Y. Murakami, F. Sugahara, J. Pascual-Anaya, R. Kusakabe, S. Higuchi, Y. Yamaguchi, W. Takagi, H. Kaiya, Y. Ishihama, S. Miyake, T. Kaku, T. Tanaka, D. Sipp, M. Tan, A. D. M. Dove, T. D. Read, D. Lagman, D. Ocampo Daza, D. Larhammar, Y. Uno and S. Mazan for insightful discussion. k, Putative Hox C gene repertories of diverse elasmobranchs. Don't be afraid of sharks any more. 2. Proc. Sci. Y.H., K.Y., K.Tatsumi, K.Tanaka, F.M., Y.K., I.K., R.Nozu, R.Nakagawa, R.M., K.M., S.H., K.O., O.N., S.D.K., C.T., M.Kadota, M.Koyanagi and S.Kuraku performed experiments. USA 105, 38193824 (2008). Of these hormone and receptor genes, all but leptin were suggested to have existed in the vertebrate ancestor by the presence of possible cyclostome orthologues or gene duplication that probably occurred in genome expansion before the divergence of all extant vertebrates33 (Fig.