{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,12]],"date-time":"2026-06-12T20:48:02Z","timestamp":1781297282523,"version":"3.54.1"},"reference-count":55,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2024,10,19]],"date-time":"2024-10-19T00:00:00Z","timestamp":1729296000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"KAKENHI from the Japan Society for the Promotion of Science","award":["23H02859"],"award-info":[{"award-number":["23H02859"]}]},{"name":"KAKENHI from the Japan Society for the Promotion of Science","award":["23K27550"],"award-info":[{"award-number":["23K27550"]}]},{"name":"OTSUCLE in Tokushima University","award":["23H02859"],"award-info":[{"award-number":["23H02859"]}]},{"name":"OTSUCLE in Tokushima University","award":["23K27550"],"award-info":[{"award-number":["23K27550"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The intestinal tract is a typical radiosensitive tissue, and radiation rectal injury is a severe side effect that limits the prescribed dose in radiotherapy of the abdominal and pelvic region. Understanding the post-irradiation kinetics of Lgr5-positive stem cells is crucial in comprehending this adverse process. In this study, we utilized Lgr5-EGFP knock-in mice expressing EGFP and LGR5 antibody fluorescence staining of wild-type mice. At the state of radiation injury, the qPCR analysis showed a significant decrease in the expression level of Lgr5 in the rectal epithelial tissue. The dose-response relationship analysis showed that at low to moderate doses up to 10 gray (Gy), Lgr5-clustered populations were observed at the base of the crypt, whereas at sublethal doses (20 Gy and 29 Gy), the cells exhibited a dot-like scatter pattern, termed Lgr5-dotted populations. During recovery, 30 days post-irradiation, Lgr5-clustered populations gradually re-emerged while Lgr5-dotted populations declined, implying that some of the Lgr5-dotted stem cell populations re-clustered, aiding regenerations. Based on statistical analysis of the dose-response relationship using wild-type mice, the threshold dose for destroying these stem cell structures is 18 Gy. These findings may help set doses in mouse abdominal irradiation experiments for radiation intestinal injury and for understanding the histological process of injury development.<\/jats:p>","DOI":"10.3390\/ijms252011252","type":"journal-article","created":{"date-parts":[[2024,10,21]],"date-time":"2024-10-21T03:57:09Z","timestamp":1729483029000},"page":"11252","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Rectal Epithelial Stem Cell Kinetics in Acute Radiation Proctitis"],"prefix":"10.3390","volume":"25","author":[{"given":"Sharmila","family":"Ghosh","sequence":"first","affiliation":[{"name":"Graduate School of Biomedical Sciences, Tokushima University, Tokushima 770-8503, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9988-6136","authenticated-orcid":false,"given":"Akinori","family":"Morita","sequence":"additional","affiliation":[{"name":"Graduate School of Biomedical Sciences, Tokushima University, Tokushima 770-8503, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8960-6110","authenticated-orcid":false,"given":"Yuichi","family":"Nishiyama","sequence":"additional","affiliation":[{"name":"Graduate School of Biomedical Sciences, Tokushima University, Tokushima 770-8503, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masahiro","family":"Sakaue","sequence":"additional","affiliation":[{"name":"Graduate School of Biomedical Sciences, Tokushima University, Tokushima 770-8503, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ken","family":"Fujiwara","sequence":"additional","affiliation":[{"name":"Graduate School of Biomedical Sciences, Tokushima University, Tokushima 770-8503, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daiki","family":"Morita","sequence":"additional","affiliation":[{"name":"Graduate School of Biomedical Sciences, Tokushima University, Tokushima 770-8503, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuichiro","family":"Sonoyama","sequence":"additional","affiliation":[{"name":"Graduate School of Biomedical Sciences, Tokushima University, Tokushima 770-8503, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuichi","family":"Higashi","sequence":"additional","affiliation":[{"name":"Graduate School of Biomedical Sciences, Tokushima University, Tokushima 770-8503, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1219-1014","authenticated-orcid":false,"given":"Megumi","family":"Sasatani","sequence":"additional","affiliation":[{"name":"Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima 754-8553, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,10,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2255","DOI":"10.1007\/s00520-013-1782-y","article-title":"Structured Gastroenterological Intervention and Improved Outcome for Patients with Chronic Gastrointestinal Symptoms Following Pelvic Radiotherapy","volume":"21","author":"Henson","year":"2013","journal-title":"Support Care Cancer"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"544","DOI":"10.1097\/COC.0000000000000320","article-title":"A Phase II Study to Prevent Radiation-Induced Rectal Injury With Lovastatin","volume":"41","author":"Anscher","year":"2018","journal-title":"Am. J. Clin. Oncol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1111","DOI":"10.1016\/S0360-3016(00)00744-6","article-title":"Acute Radiation Proctitis: A Sequential Clinicopathologic Study during Pelvic Radiotherapy","volume":"48","author":"Hovdenak","year":"2000","journal-title":"Int. J. Radiat. Oncol. Biol. Phys."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1055\/s-2007-970202","article-title":"Radiation Colitis and Proctitis","volume":"20","author":"Kennedy","year":"2007","journal-title":"Clin. Colon Rectal Surg."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1055\/s-0035-1547337","article-title":"Radiation Proctopathy","volume":"28","author":"Grodsky","year":"2015","journal-title":"Clin. Colon Rectal Surg."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1186\/s13287-017-0763-3","article-title":"BCN057 Induces Intestinal Stem Cell Repair and Mitigates Radiation-Induced Intestinal Injury","volume":"9","author":"Bhanja","year":"2018","journal-title":"Stem. Cell Res. Ther."},{"key":"ref_7","first-page":"10","article-title":"Molecular Mechanisms of Acute Radiation Intestinal Injury and Its Control","volume":"13","author":"Morita","year":"2024","journal-title":"Radiat. Environ. Med."},{"key":"ref_8","first-page":"4879","article-title":"Histopathological Comparison of Topical Therapy Modalities for Acute Radiation Proctitis in an Experimental Rat Model","volume":"12","author":"Korkut","year":"2006","journal-title":"World J. Gastroenterol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"917941","DOI":"10.1155\/2011\/917941","article-title":"Radiation Proctitis: Current Strategies in Management","volume":"2011","author":"Do","year":"2011","journal-title":"Gastroenterol. Res. Pract."},{"key":"ref_10","first-page":"587","article-title":"New Strategies in Rectal Cancer","volume":"97","author":"Vailati","year":"2017","journal-title":"Surg. Clin."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1016\/j.semradonc.2011.02.010","article-title":"Nonoperative Approaches to Rectal Cancer: A Critical Evaluation","volume":"21","author":"Perez","year":"2011","journal-title":"Semin. Radiat. Oncol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"3639","DOI":"10.1242\/dev.133132","article-title":"Regulation and Plasticity of Intestinal Stem Cells during Homeostasis and Regeneration","volume":"143","author":"Beumer","year":"2016","journal-title":"Development"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"340","DOI":"10.1007\/s11894-010-0130-3","article-title":"Intestinal Stem Cells","volume":"12","author":"Umar","year":"2010","journal-title":"Curr. Gastroenterol. Rep."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"262","DOI":"10.1038\/nature07935","article-title":"Single Lgr5 Stem Cells Build Crypt-Villus Structures in Vitro without a Mesenchymal Niche","volume":"459","author":"Sato","year":"2009","journal-title":"Nature"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"185","DOI":"10.3748\/wjg.v19.i2.185","article-title":"Gastrointestinal Radiation Injury: Symptoms, Risk Factors and Mechanisms","volume":"19","author":"Shadad","year":"2013","journal-title":"World J. Gastroenterol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"452","DOI":"10.1016\/j.stem.2012.09.009","article-title":"Identifying the Stem Cell of the Intestinal Crypt: Strategies and Pitfalls","volume":"11","author":"Barker","year":"2012","journal-title":"Cell Stem. Cell"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1266","DOI":"10.1053\/j.gastro.2012.07.106","article-title":"Crypt Base Columnar Stem Cells in Small Intestines of Mice Are Radioresistant","volume":"143","author":"Hua","year":"2012","journal-title":"Gastroenterology"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1003","DOI":"10.1038\/nature06196","article-title":"Identification of Stem Cells in Small Intestine and Colon by Marker Gene Lgr5","volume":"449","author":"Barker","year":"2007","journal-title":"Nature"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"13096","DOI":"10.1038\/ncomms13096","article-title":"Macrophage-Derived Extracellular Vesicle-Packaged WNTs Rescue Intestinal Stem Cells and Enhance Survival after Radiation Injury","volume":"7","author":"Saha","year":"2016","journal-title":"Nat. Commun."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"380","DOI":"10.1016\/j.tcb.2018.01.010","article-title":"Recent Advances in Lgr5+ Stem Cell Research","volume":"28","author":"Leung","year":"2018","journal-title":"Trends Cell Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1038\/nrgastro.2014.46","article-title":"Radiation Enteropathy\u2014Pathogenesis, Treatment and Prevention","volume":"11","author":"Denham","year":"2014","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"244","DOI":"10.1186\/1477-7819-10-244","article-title":"Lgr5 Is a Potential Marker of Colorectal Carcinoma Stem Cells That Correlates with Patient Survival","volume":"10","author":"Wu","year":"2012","journal-title":"World J. Surg. Oncol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1016\/j.biopha.2014.03.016","article-title":"Expression of Lgr5, a Marker of Intestinal Stem Cells, in Colorectal Cancer and Its Clinicopathological Significance","volume":"68","author":"He","year":"2014","journal-title":"Biomed. Pharmacother."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1007\/s00384-010-0903-z","article-title":"Expression of Lgr5 in Human Colorectal Carcinogenesis and Its Potential Correlation with \u03b2-Catenin","volume":"25","author":"Fan","year":"2010","journal-title":"Int. J. Colorectal Dis."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1410","DOI":"10.1038\/s41416-018-0118-6","article-title":"Targeting LGR5 in Colorectal Cancer: Therapeutic Gold or Too Plastic?","volume":"118","author":"Morgan","year":"2018","journal-title":"Br. J. Cancer"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"414","DOI":"10.3109\/02841867809128251","article-title":"Radiation Effects on the Fine Blood Vessels in Abdominal Organs of Mice","volume":"17","author":"Egawa","year":"1978","journal-title":"Acta Radiol. Oncol. Radiat. Phys. Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"682","DOI":"10.3346\/jkms.2000.15.6.682","article-title":"A Rat Model for Radiation-Induced Proctitis","volume":"15","author":"Kang","year":"2000","journal-title":"J. Korean Med. Sci."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1559325820956783","DOI":"10.1177\/1559325820956783","article-title":"At What Dose Can Total Body and Whole Abdominal Irradiation Cause Lethal Intestinal Injury Among C57BL\/6J Mice?","volume":"18","author":"Gu","year":"2020","journal-title":"Dose-Response"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"S\u00e9mont, A., Demarquay, C., Bessout, R., Durand, C., Benderitter, M., and Mathieu, N. (2013). Mesenchymal Stem Cell Therapy Stimulates Endogenous Host Progenitor Cells to Improve Colonic Epithelial Regeneration. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0070170"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1053\/j.gastro.2009.05.035","article-title":"OLFM4 Is a Robust Marker for Stem Cells in Human Intestine and Marks a Subset of Colorectal Cancer Cells","volume":"137","author":"Haegebarth","year":"2009","journal-title":"Gastroenterology"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"903","DOI":"10.1016\/j.cell.2009.01.031","article-title":"Transcription Factor Achaete Scute-Like 2 Controls Intestinal Stem Cell Fate","volume":"136","author":"Hatzis","year":"2009","journal-title":"Cell"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/j.stem.2015.01.014","article-title":"Ascl2 Reinforces Intestinal Stem Cell Identity","volume":"16","author":"Yan","year":"2015","journal-title":"Cell Stem. Cell"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Nishiyama, K., Sugiyama, M., and Mukai, T. (2016). Adhesion Properties of Lactic Acid Bacteria on Intestinal Mucin. Microorganisms, 4.","DOI":"10.3390\/microorganisms4030034"},{"key":"ref_34","first-page":"261","article-title":"Microcolony Survival Assay for Cells of Mouse Intestinal Mucosa Exposed to Radiation","volume":"17","author":"Withers","year":"1970","journal-title":"Int. J. Radiat. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"598","DOI":"10.2307\/3572619","article-title":"Radiosensitivity and Fractionation Response of Crypt Cells of Mouse Jejunum","volume":"38","author":"Withers","year":"1969","journal-title":"Radiat. Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1111\/codi.12507","article-title":"Sucralfate Paste Enema: A New Method of Topical Treatment for Haemorrhagic Radiation Proctitis","volume":"16","author":"McElvanna","year":"2014","journal-title":"Color. Dis."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1124","DOI":"10.1016\/j.ijrobp.2007.11.044","article-title":"Incidence of Late Rectal and Urinary Toxicities After Three-Dimensional Conformal Radiotherapy and Intensity-Modulated Radiotherapy for Localized Prostate Cancer","volume":"70","author":"Zelefsky","year":"2008","journal-title":"Int. J. Radiat. Oncol. Biol. Phys."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1232","DOI":"10.1016\/S0140-6736(00)02787-2","article-title":"Topical Butyrate for Acute Radiation Proctitis: Randomised, Crossover Trial","volume":"356","author":"Vernia","year":"2000","journal-title":"Lancet"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"100067","DOI":"10.1016\/j.arres.2023.100067","article-title":"Effects of X-Ray Irradiation on the Redox Status of Mouse Colorectal Tissues","volume":"8","author":"Ueno","year":"2023","journal-title":"Adv. Redox Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"821","DOI":"10.1098\/rstb.1998.0246","article-title":"Stem Cells in Gastrointestinal Epithelium: Numbers, Characteristics and Death","volume":"353","author":"Potten","year":"1998","journal-title":"Philos. Trans. R Soc. Lond. B Biol. Sci."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"637","DOI":"10.1667\/RR3253.1","article-title":"Ionizing Radiation Leads to the Replacement and de Novo Production of Colonic Lgr5 Stem Cells","volume":"179","author":"Otsuka","year":"2013","journal-title":"Radiat. Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"993","DOI":"10.1038\/bjc.1998.618","article-title":"The Relationship between Ionizing Radiation-Induced Apoptosis and Stem Cells in the Small and Large Intestine","volume":"78","author":"Potten","year":"1998","journal-title":"Br. J. Cancer"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1046\/j.1365-2184.2003.00279.x","article-title":"Intestinal Crypt Clonogens: A New Interpretation of Radiation Survival Curve Shape and Clonogenic Cell Number","volume":"36","author":"Roberts","year":"2003","journal-title":"Cell Prolif."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1097\/HP.0b013e318266ee13","article-title":"Acute Gastrointestinal Syndrome in High-Dose Irradiated Mice","volume":"103","author":"Booth","year":"2012","journal-title":"Health Phys."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1016\/j.stemcr.2016.01.014","article-title":"A Versatile Strategy for Isolating a Highly Enriched Population of Intestinal Stem Cells","volume":"6","author":"Nefzger","year":"2016","journal-title":"Stem Cell Rep."},{"key":"ref_46","first-page":"1373","article-title":"Single Cell Time-Lapse Imaging of Focus Formation by the DNA Damage-Response Protein 53BP1 after UVC Irradiation of Human Pancreatic Cancer Cells","volume":"33","author":"Miwa","year":"2013","journal-title":"Anticancer Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1381","DOI":"10.1083\/jcb.151.7.1381","article-title":"p53 Binding Protein 1 (53bp1) Is an Early Participant in the Cellular Response to DNA Double-Strand Breaks","volume":"151","author":"Schultz","year":"2000","journal-title":"J. Cell Biol."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Sollazzo, A., Brzozowska, B., Cheng, L., Lundholm, L., Scherthan, H., and Wojcik, A. (2018). Live Dynamics of 53BP1 Foci Following Simultaneous Induction of Clustered and Dispersed DNA Damage in U2OS Cells. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19020519"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1073\/pnas.1117849108","article-title":"Evidence for Formation of DNA Repair Centers and Dose-Response Nonlinearity in Human Cells","volume":"109","author":"Neumaier","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"e2119132119","DOI":"10.1073\/pnas.2119132119","article-title":"Formation of Clustered DNA Damage in Vivo upon Irradiation with Ionizing Radiation: Visualization and Analysis with Atomic Force Microscopy","volume":"119","author":"Nakano","year":"2022","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1063","DOI":"10.21037\/atm-20-4542","article-title":"Regulation of the Regeneration of Intestinal Stem Cells after Irradiation","volume":"8","author":"Liang","year":"2020","journal-title":"Ann. Transl. Med."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"627","DOI":"10.1016\/j.stem.2015.04.013","article-title":"Krt19+\/Lgr5\u2212 Cells Are Radioresistant Cancer-Initiating Stem Cells in the Colon and Intestine","volume":"16","author":"Asfaha","year":"2015","journal-title":"Cell Stem Cell"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1241","DOI":"10.1080\/09553002.2021.1941377","article-title":"Evaluation of Sodium Orthovanadate as a Radioprotective Agent under Total-Body Irradiation and Partial-Body Irradiation Conditions in Mice","volume":"97","author":"Nishiyama","year":"2021","journal-title":"Int. J. Radiat. Biol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1258\/002367781780958577","article-title":"The \u2018Swiss Roll\u2019: A Simple Technique for Histological Studies of the Rodent Intestine","volume":"15","author":"Moolenbeek","year":"1981","journal-title":"Lab. Anim."},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Morita, A., Ochi, S., Satoh, H., Ujita, S., Matsushita, Y., Tada, K., Toyoda, M., Nishiyama, Y., Mizuno, K., and Deguchi, Y. (2021). A Novel RNA Synthesis Inhibitor, STK160830, Has Negligible DNA-Intercalating Activity for Triggering A p53 Response and Can Inhibit p53-Dependent Apoptosis. Life, 11.","DOI":"10.3390\/life11101087"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/20\/11252\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:16:41Z","timestamp":1760113001000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/20\/11252"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,10,19]]},"references-count":55,"journal-issue":{"issue":"20","published-online":{"date-parts":[[2024,10]]}},"alternative-id":["ijms252011252"],"URL":"https:\/\/doi.org\/10.3390\/ijms252011252","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,10,19]]}}}