{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T22:01:18Z","timestamp":1791237678623,"version":"4.1.0"},"reference-count":51,"publisher":"Zoological Society of Japan","issue":"1","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Zoological Science"],"published-print":{"date-parts":[[2025,12,18]]},"abstract":"<jats:p>Deuterostomes generally share a functional digestive system involving the sequential steps of ingestion, digestion, absorption, and elimination along their one-way gut. Previous gene expression studies on pancreas-related exocrine digestive enzymes (XDEs), pancreas-related transcription factors (TFs), and ParaHox genes in the ascidian Ciona robusta showed non-contiguous and multi-regional pancreatic features along the Ciona gut. The combination of classical in situ hybridization and recent RNA-seq techniques in lancelets has allowed the fundamental features of the gut in basal chordates (lancelets and ascidians) to be examined in more detail. In the present study, we investigated the spatial expression of pancreas-related XDE and TF genes with a focus on their regionality in the adult lancelet gut. Whole-mount in situ hybridization using Branchiostoma japonicum revealed non-contiguous and multiple pancreatic regions in the lancelet gut. Public bulk RNA-seq data on lancelets (Branchiostoma belcheri, B. floridae, and B. lanceolatum) also supported the multi-regional distribution of pancreatic features along the lancelet gut. In addition, single-cell RNA-seq data indicated that the lancelet hepatic caecum comprised multiple cell clusters with the differential gene expression of pancreas-related XDEs, Pdx, and Cdx. Gene knockout experiments on ascidian Pdx suggested that \u201cthe multi-regional expression of pancreas-related XDE genes\u201d in basal chordates is associated with Pdx-independent regulation.<\/jats:p>","DOI":"10.2108\/zs250035","type":"journal-article","created":{"date-parts":[[2025,12,17]],"date-time":"2025-12-17T02:20:39Z","timestamp":1765938039000},"page":"106-117","source":"Crossref","is-referenced-by-count":1,"title":["Non-Contiguous and Multi-Regional Expression of Pancreas-Related Genes along the Lancelet Gut"],"prefix":"10.2108","volume":"43","author":[{"given":"Haruka","family":"Fujiwara","sequence":"first","affiliation":[{"name":"Department of Biology, Graduate School of Science, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba 263-8522, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Satoshi","family":"Nakayama","sequence":"additional","affiliation":[{"name":"Department of Biology, Graduate School of Science, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba 263-8522, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rin","family":"Iguchi","sequence":"additional","affiliation":[{"name":"Department of Biology, Graduate School of Science, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba 263-8522, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Toshio","family":"Sekiguchi","sequence":"additional","affiliation":[{"name":"The Noto Marine Laboratory, Division of Marine Environmental Studies, Institute of Nature and Environmental Technology, Kanazawa University, Housu-gun, Ishikawa 927-0553, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasunori","family":"Sasakura","sequence":"additional","affiliation":[{"name":"Shimoda Marine Research Center, University of Tsukuba, 5-10-1 Shimoda, Shizuoka 415-0025, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masato","family":"Kiyomoto","sequence":"additional","affiliation":[{"name":"Tateyama Marine Laboratory, Institute for Marine and Coastal Research, Ochanomizu University, Tateyama, Chiba 294-0301, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michio","family":"Ogasawara","sequence":"additional","affiliation":[{"name":"Department of Biology, Graduate School of Science, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba 263-8522, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"848","reference":[{"key":"1","doi-asserted-by":"crossref","unstructured":"R Annunziata, C Andrikou, M Perillo, et al.  \"Cell Tissue Res.\"  377, \"Development and evolution of gut structures: from molecules to function.\" 445 (2019)","DOI":"10.1007\/s00441-019-03093-9"},{"key":"2","doi-asserted-by":"crossref","unstructured":"R Annunziata, P Martinez, MI Arnone,  \"BMC Biol.\"  11, \"Intact cluster and chordate-like expression of ParaHox genes in a sea star.\" 68 (2013)","DOI":"10.1186\/1741-7007-11-68"},{"key":"3","doi-asserted-by":"crossref","unstructured":"S Ashizawa, FC Brunicardi, XP Wang,  \"Pancreas.\"  28, \"PDX-1 and the pancreas.\" 109 (2004)","DOI":"10.1097\/00006676-200403000-00001"},{"key":"4","doi-asserted-by":"crossref","unstructured":"EJW Barrington,  \"Phil Trans R Soc Lond.\"  228, \"The digestive system of Amphioxus (Branchiostoma) Lanceolatus.\" 269 (1937)","DOI":"10.1098\/rstb.1937.0013"},{"key":"5","doi-asserted-by":"crossref","unstructured":"SJ Bourlat, T Juliusdottir, CJ Lowe, et al.  \"Nature.\"  444, \"Deuterostome phylogeny reveals monophyletic chordates and the new phylum Xenoturbellida.\" 85 (2006)","DOI":"10.1038\/nature05241"},{"key":"6","doi-asserted-by":"crossref","unstructured":"NM Brooke, J Garcia-Fern\u00e0ndez, PWH Holland,  \"Nature.\"  392, \"The ParaHox gene cluster is an evolutionary sister of the Hox gene cluster.\" 920 (1998)","DOI":"10.1038\/31933"},{"key":"8","doi-asserted-by":"crossref","unstructured":"JC Corbo, M Levine, RW Zeller,  \"Development.\"  124, \"Characterization of a notochord-specific enhancer from the Brachyury promoter region of the ascidian, Ciona intestinalis.\" 589 (1997)","DOI":"10.1242\/dev.124.3.589"},{"key":"9","doi-asserted-by":"crossref","unstructured":"Y Dai, R Pan, Q Pan, et al.  \"Sci Adv.\"  10, \"Single-cell profiling of the amphioxus digestive tract reveals conservation of endocrine cells in chordates.\" eadq0702 (2024)","DOI":"10.1126\/sciadv.adq0702"},{"key":"11","doi-asserted-by":"crossref","unstructured":"W Feng  and S Zhang  \"Mol Biol Rep.\"  39, \"A trypsin homolog in amphioxus: expression, enzymatic activity and evolution.\" 1745 (2012)","DOI":"10.1007\/s11033-011-0915-y"},{"key":"12","doi-asserted-by":"crossref","unstructured":"AC Fr\u00f6bius  and EC Seaver  \"Dev Genes Evol.\"  216, \"ParaHox gene expression in the polychaete annelid Capitella sp. I.\" 81 (2006)","DOI":"10.1007\/s00427-005-0049-0"},{"key":"13","doi-asserted-by":"crossref","unstructured":"A Grapin-Botton  and DA Melton  \"Trends Genet.\"  16, \"Endoderm development: from patterning to organogenesis.\" 124 (2000)","DOI":"10.1016\/S0168-9525(99)01957-5"},{"key":"14","doi-asserted-by":"crossref","unstructured":"LZ Holland, TV Venkatesh, A Gorlin, et al.  \"Dev Genes Evol.\"  208, \"Characterization and developmental expression of AmphiNk2-2, an NK2 class homeobox gene from Amphioxus (Phylum Chordata; Subphylum Cephalochordata).\" 100 (1998)","DOI":"10.1007\/s004270050159"},{"key":"15","doi-asserted-by":"crossref","unstructured":"ND Holland, LZ Holland, PWH Holland,  \"Nature.\"  520, \"Scenarios for the making of vertebrates.\" 450 (2015)","DOI":"10.1038\/nature14433"},{"key":"16","doi-asserted-by":"crossref","unstructured":"SR Holmstrom, T Deering, GH Swift, et al.  \"Genes Dev.\"  25, \"LRH-1 and PTF1-L coregulate an exocrine pancreas-specific transcriptional network for digestive function.\" 1674 (2011)","DOI":"10.1101\/gad.16860911"},{"key":"17","doi-asserted-by":"crossref","unstructured":"Z Huang, L Xu, C Cai, et al.  \"Proc Natl Acad Sci U S A.\"  120, \"Three amphioxus reference genomes reveal gene and chromosome evolution of chordates.\" e2201504120 (2023)","DOI":"10.1073\/pnas.2201504120"},{"key":"18","doi-asserted-by":"crossref","unstructured":"H Hui  and R Perfetti  \"Eur J Endocrinol.\"  146, \"Pancreas duodenum homeobox-1 regulates pancreas development during embryogenesis and islet cell function in adulthood.\" 129 (2002)","DOI":"10.1530\/eje.0.1460129"},{"key":"19","doi-asserted-by":"crossref","unstructured":"R Iguchi, S Nakayama, Y Sasakura, et al.  \"Cell Tissue Res.\"  394, \"Repetitive and zonal expression profiles of absorption-related genes in the gastrointestinal tract of ascidian Ciona intestinalis type A.\" 343 (2023)","DOI":"10.1007\/s00441-023-03828-9"},{"key":"20","doi-asserted-by":"crossref","unstructured":"R Iguchi, K Usui, S Nakayama, et al.  \"Cell Tissue Res.\"  394, \"Multi-regional expression of pancreas-related digestive enzyme genes in the intestinal chamber of the ascidian Ciona intestinalis type A.\" 423 (2023)","DOI":"10.1007\/s00441-023-03839-6"},{"key":"21","doi-asserted-by":"crossref","unstructured":"T Ikuta, YC Chen, R Annunziata, et al.  \"BMC Evol Biol.\"  13, \"Identification of an intact ParaHox cluster with temporal colinearity but altered spatial colinearity in the hemichordate Ptychodera flava.\" 129 (2013)","DOI":"10.1186\/1471-2148-13-129"},{"key":"23","doi-asserted-by":"crossref","unstructured":"C Lecroisey, Y Le P\u00e9tillon, H Escriva, et al.  \"PLOS ONE.\"  10, \"Identification, evolution and expression of an insulin-like peptide in the cephalochordate Branchiostoma lanceolatum.\" e0119461 (2015)","DOI":"10.1371\/journal.pone.0119461"},{"key":"25","doi-asserted-by":"crossref","unstructured":"H Li, Z Gao, S Zhang,  \"Tissue Cell.\"  79, \"Localization of trypsin, chymotrypsin and elastase in the digestive tract of amphioxus Branchiostoma japonicum with implications to the origin of vertebrate pancreas.\" 101943 (2022)","DOI":"10.1016\/j.tice.2022.101943"},{"key":"26","doi-asserted-by":"crossref","unstructured":"LC Li, WL Qiu, YW Zhang, et al.  \"EMBO Rep.\"  19, \"Single-cell transcriptomic analyses reveal distinct dorsal\/ ventral pancreatic programs.\" e46148 (2018)","DOI":"10.15252\/embr.201846148"},{"key":"27","doi-asserted-by":"crossref","unstructured":"CJ Lowe, DN Clarke, DM Medeiros, et al.  \"Nature.\"  520, \"The deuterostome context of chordate origins.\" 456 (2015)","DOI":"10.1038\/nature14434"},{"key":"28","unstructured":"F Marl\u00e9taz, PN Firbas, I Maeso, et al.  \"Nature.\"  564, \"Amphioxus functional genomics and the origins of vertebrate gene regulation.\" 64 (2018)"},{"key":"30","doi-asserted-by":"crossref","unstructured":"V Mehta, PE Hopson, Y Smadi, et al.  \"Front Pediatr.\"  10, \"Development of the human pancreas and its exocrine function.\" 909648 (2022)","DOI":"10.3389\/fped.2022.909648"},{"key":"31","doi-asserted-by":"crossref","unstructured":"D Meulemans  and M Bronner-Fraser  \"Development.\"  129, \"Amphioxus and lamprey AP-2 genes: implications for neural crest evolution and migration patterns.\" 4953 (2002)","DOI":"10.1242\/dev.129.21.4953"},{"key":"32","doi-asserted-by":"crossref","unstructured":"S Nakayama  and M Ogasawara  \"Cell Tissue Res.\"  370, \"Compartmentalized expression patterns of pancreatic- and gastric-related genes in the alimentary canal of the ascidian Ciona intestinalis: evolutionary insights into the functional regionality of the gastrointestinal tract in Olfactores.\" 113 (2017)","DOI":"10.1007\/s00441-017-2627-7"},{"key":"33","doi-asserted-by":"crossref","unstructured":"S Nakayama, K Satou, W Orito, et al.  \"Cell Tissue Res.\"  365, \"Ordered expression pattern of Hox and ParaHox genes along the alimentary canal in the ascidian juvenile.\" 65 (2016)","DOI":"10.1007\/s00441-016-2360-7"},{"key":"34","doi-asserted-by":"crossref","unstructured":"MF Offield, TL Jetton, PA Labosky, et al.  \"Development.\"  122, \"PDX-1 is required for pancreatic outgrowth and differentiation of the rostral duodenum.\" 983 (1996)","DOI":"10.1242\/dev.122.3.983"},{"key":"35","doi-asserted-by":"crossref","unstructured":"M Ogasawara, T Minokawa, Y Sasakura, et al.  \"Zoolog Sci.\"  18, \"A large-scale whole-mount in situ hybridization system: rapid one-tube preparation of DIG-labeled RNA probes and high throughput hybridization using 96-well silent screen plates.\" 187 (2001)","DOI":"10.2108\/zsj.18.187"},{"key":"36","doi-asserted-by":"crossref","unstructured":"W Orito, F Ohhira, M Ogasawara,  \"Cell Tissue Res.\"  362, \"Gene expression profiles of FABP genes in protochordates, Ciona intestinalis and Branchiostoma belcheri.\" 331 (2015)","DOI":"10.1007\/s00441-015-2198-4"},{"key":"38","doi-asserted-by":"crossref","unstructured":"T Sakuma, H Ochiai, T Kaneko, et al.  \"Sci Rep.\"  3, \"Repeating pattern of non-RVD variations in DNA-binding modules enhances TALEN activity.\" 3379 (2013)","DOI":"10.1038\/srep03379"},{"key":"39","doi-asserted-by":"crossref","unstructured":"Y Sasakura, Y Ogura, N Treen, et al.  \"Proc R Soc B.\"  283, \"Transcriptional regulation of a horizontally transferred gene from bacterium to chordate.\" 20161712 (2016)","DOI":"10.1098\/rspb.2016.1712"},{"key":"40","doi-asserted-by":"crossref","unstructured":"J Suckale  and M Solimena  \"Front Biosci.\"  13, \"Pancreas islets in metabolic signaling\u2013focus on the beta-cell.\" 7156 (2008)","DOI":"10.2741\/3218"},{"key":"41","doi-asserted-by":"crossref","unstructured":"T Takeuchi, R Koyanagi, F Gyoja, et al.  \"Zool Lett.\"  2, \"Bivalve-specific gene expansion in the pearl oyster genome: implications of adaptation to a sessile lifestyle.\" 3 (2016)","DOI":"10.1186\/s40851-016-0039-2"},{"key":"42","doi-asserted-by":"crossref","unstructured":"EN Temereva  and RP Kostyuchenko  \"J Exp Zool B.\"  334, \"Digestive system development and posterior Hox\/Parahox gene expression during larval life and metamorphosis of the Phoronid Phoronopsis harmeri.\" 136 (2025)","DOI":"10.1002\/jez.b.23286"},{"key":"43","doi-asserted-by":"crossref","unstructured":"N Treen, K Yoshida, T Sakuma, et al.  \"Development.\"  141, \"Tissue-specific and ubiquitous gene knockouts by TALEN electroporation provide new approaches to investigating gene function in Ciona.\" 481 (2014)","DOI":"10.1242\/dev.099572"},{"key":"44","doi-asserted-by":"crossref","unstructured":"B Wu, W Xu, K Wu, et al.  \"Nat Ecol Evol.\"  8, \"Single-cell analysis of the amphioxus hepatic caecum and vertebrate liver reveals genetic mechanisms of vertebrate liver evolution.\" 1972 (2024)","DOI":"10.1038\/s41559-024-02510-9"},{"key":"45","doi-asserted-by":"crossref","unstructured":"K Yasui, T Igawa, T Kaji, et al.  \"J Exp Zool B.\"  320, \"Stable aquaculture of the Japanese lancelet Branchiostoma japonicum for 7 years.\" 538 (2013)","DOI":"10.1002\/jez.b.22540"},{"key":"46","doi-asserted-by":"crossref","unstructured":"K Yoshida, A Nakahata, N Treen, et al.  \"Development.\"  144, \"Hox-mediated endodermal identity patterns pharyngeal muscle formation in the chordate pharynx.\" 1629 (2017)","DOI":"10.1242\/dev.144436"},{"key":"47","doi-asserted-by":"crossref","unstructured":"L You, J Chi, S Huang, et al.  \"Database (Oxford).\"  2019, \"LanceletDB: an integrated genome database for lancelet, comparing domain types and combination in orthologues among lancelet and other species.\" baz056 (2019)","DOI":"10.1093\/database\/baz056"},{"key":"48","doi-asserted-by":"crossref","unstructured":"JK Yu, Y Satou, ND Holland, et al.  \"Nature.\"  445, \"Axial patterning in cephalochordates and the evolution of the organizer.\" 613 (2007)","DOI":"10.1038\/nature05472"},{"key":"49","doi-asserted-by":"crossref","unstructured":"E Zecchin, A Mavropoulos, N Devos, et al.  \"Dev Biol.\"  268, \"Evolutionary conserved role of ptf1a in the specification of exocrine pancreatic fates.\" 174 (2004)","DOI":"10.1016\/j.ydbio.2003.12.016"},{"key":"50","doi-asserted-by":"crossref","unstructured":"S Zhang, Z Shen, H Li,  \"Acta Oceanol Sin.\"  42, \"Hepatic caecum of amphioxus and origin of vertebrate liver.\" 1 (2023)","DOI":"10.1007\/s13131-023-2259-6"},{"key":"51","doi-asserted-by":"crossref","unstructured":"Y Zhong, C Herrera-\u00dabeda, J Garcia-Fern\u00e0ndez, et al.  \"BMC Biol.\"  18, \"Mutation of amphioxus Pdx and Cdx demonstrates conserved roles for ParaHox genes in gut, anus and tail patterning.\" 68 (2020)","DOI":"10.1186\/s12915-020-00796-2"},{"key":"22","doi-asserted-by":"crossref","unstructured":"D Kim, JM Paggi, C Park, et al.  \"Nat Biotechnol.\"  37, \"Graph-based genome alignment and genotyping with HISAT2 and HISAT-genotype.\" 907 (2019)","DOI":"10.1038\/s41587-019-0201-4"},{"key":"10","doi-asserted-by":"crossref","unstructured":"F Delsuc, H Brinkmann, D Chourrout, et al.  \"Nature.\"  439, \"Tunicates and not cephalochordates are the closest living relatives of vertebrates.\" 965 (2006)","DOI":"10.1038\/nature04336"},{"key":"37","unstructured":"NH Putnam, T Butts, DE Ferrier, et al.  \"Nature.\"  453, \"The amphioxus genome and the evolution of the chordate karyotype.\" 1064 (2008)"},{"key":"24","doi-asserted-by":"crossref","unstructured":"B Li  and CN Dewey  \"BMC Bioinformatics.\"  12, \"RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome.\" 323 (2011)","DOI":"10.1186\/1471-2105-12-323"},{"key":"7","doi-asserted-by":"crossref","unstructured":"T Cermak, EL Doyle, M Christian, et al.  \"Nucleic Acids Res.\"  39, \"Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting.\" e82 (2011)","DOI":"10.1093\/nar\/gkr218"},{"key":"29","doi-asserted-by":"crossref","unstructured":"S Matsubara, T Osugi, A Shiraishi, et al.  \"PLOS ONE.\"  16, \"Comparative analysis of transcriptomic profiles among ascidians, zebrafish, and mice: Insights from tissue-specific gene expression.\" e0254308 (2021)","DOI":"10.1371\/journal.pone.0254308"}],"container-title":["Zoological Science"],"original-title":[],"deposited":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T21:36:52Z","timestamp":1791236212000},"score":1,"resource":{"primary":{"URL":"https:\/\/bioone.org\/journals\/zoological-science\/volume-43\/issue-1\/zs250035\/Non-Contiguous-and-Multi-Regional-Expression-of-Pancreas-Related-Genes\/10.2108\/zs250035.full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,18]]},"references-count":51,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,2,1]]}},"URL":"https:\/\/doi.org\/10.2108\/zs250035","relation":{},"ISSN":["0289-0003","2212-3830"],"issn-type":[{"value":"0289-0003","type":"print"},{"value":"2212-3830","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,18]]}}}