{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,1]],"date-time":"2026-08-01T10:03:38Z","timestamp":1785578618193,"version":"3.56.0"},"reference-count":25,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2022,3,26]],"date-time":"2022-03-26T00:00:00Z","timestamp":1648252800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Veterinary Medicine &amp;amp; Sci"],"published-print":{"date-parts":[[2022,7]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Canine histiocytic sarcoma (HS) is an aggressive cancer with morphologically variable features; therefore, obtaining a definitive diagnosis can be challenging. Two proteins, IBA\u20101, ionised calcium\u2010binding adapter molecule 1, and CD204, a macrophage scavenger receptor, have been shown to be specific immunohistochemical markers helpful in distinguishing HS from other tumour types with similar morphological features.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objectives<\/jats:title><jats:p>This study was performed to demonstrate the use of RNA in situ hybridisation (ISH) technology allowing single\u2010molecule RNA visualisation in formalin\u2010fixed paraffin\u2010embedded (FFPE) tissues as a molecular tool for the diagnosis of canine HS.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Reverse transcription polymerase chain reaction (RT\u2010PCR) and western blot analysis for IBA\u20101 and CD204 were performed to correlate gene expression and protein expression of these two markers in the histiocytic sarcoma DH82 cell line. RNA\u2010ISH for IBA\u20101 and CD204 was performed on the DH82 cell line to validate the RNA\u2010ISH probes. RNA\u2010ISH and immunohistochemistry (IHC) were performed in clinical HS FFPE samples to demonstrate mRNA and protein expression of IBA\u20101 and CD204. FFPE archived samples of canine round cell tumours, melanoma and anaplastic sarcoma were used as negative controls.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>RNA\u2010ISH and IHC showed moderate to strong expression for IBA\u20101 and CD204 in the neoplastic cells in both the canine DH82 cell line and the archived canine HS samples. RNA\u2010ISH and IHC showed scattered positive staining in the control tumours samples, consistent with macrophagic infiltration.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>RNA\u2010ISH for CD204 and IBA\u20101 appeared to have a high specificity and sensitivity in our samples and may be an additional valuable diagnostic technique in identifying HS.<\/jats:p><\/jats:sec>","DOI":"10.1002\/vms3.795","type":"journal-article","created":{"date-parts":[[2022,3,26]],"date-time":"2022-03-26T21:23:56Z","timestamp":1648329836000},"page":"1400-1408","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["RNA in situ hybridisation as a molecular diagnostic technique targeting IBA\u20101 and CD204 in canine histiocytic sarcoma"],"prefix":"10.1002","volume":"8","author":[{"given":"Julia L","family":"Engelien","sequence":"first","affiliation":[{"name":"Department of Small Animal Clinical Sciences and Comparative Diagnostic and Population Medicine College of Veterinary Medicine University of Florida  Gainesville Florida"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amandine T","family":"Lejeune","sequence":"additional","affiliation":[{"name":"Department of Small Animal Clinical Sciences and Comparative Diagnostic and Population Medicine College of Veterinary Medicine University of Florida  Gainesville Florida"},{"name":"Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California, One Shields Avenue, Davis, CA 95616."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael J","family":"Dark","sequence":"additional","affiliation":[{"name":"Department of Small Animal Clinical Sciences and Comparative Diagnostic and Population Medicine College of Veterinary Medicine University of Florida  Gainesville Florida"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rowan J","family":"Milner","sequence":"additional","affiliation":[{"name":"Department of Small Animal Clinical Sciences and Comparative Diagnostic and Population Medicine College of Veterinary Medicine University of Florida  Gainesville Florida"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keijiro","family":"Shiomitsu","sequence":"additional","affiliation":[{"name":"Department of Small Animal Clinical Sciences and Comparative Diagnostic and Population Medicine College of Veterinary Medicine University of Florida  Gainesville Florida"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2022,3,26]]},"reference":[{"key":"e_1_2_11_2_1","unstructured":"https:\/\/www.Mbio.Ncsu.Edu\/BioEdit\/Bioedit.Html"},{"key":"e_1_2_11_3_1","unstructured":"A Guide for RNAscope Data Analysis: Advanced Cell Diagnosis Manual(2021). 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