{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T20:59:31Z","timestamp":1790283571749,"version":"4.1.0"},"reference-count":45,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2025,1,16]],"date-time":"2025-01-16T00:00:00Z","timestamp":1736985600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Acta Physiologica"],"published-print":{"date-parts":[[2025,2]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Aim<\/jats:title>\n                    <jats:p>Tissue clearance is a rapidly evolving technology that allows for the three\u2010dimensional imaging of intact biological tissues. Preexisting tissue\u2010clearing techniques, such as Passive Clarity Technique (PACT) and Clear Unobstructed Brain Imaging Cocktails and Computational Analysis (CUBIC), clear tissues adequately but have distinct disadvantages, such as taking extensive time to clear tissues and degradation of endogenous tissue fluorescence. We developed a new tissue\u2010clearing technique combining PACT and CUBIC protocols to map the neural lineages expressing the transient receptor potential vanilloid type 1 (TRPV1) receptor.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>To test the effectiveness of this modified protocol, a TdTomato reporter mouse line was crossed with a separate mouse line containing Cre recombinase under the control of the TRPV1 promoter, which would result in TRPV1 cell lineages expressing green fluorescence protein (GFP).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Compared to the PACT protocol that requires several weeks to months for tissue clearance, our approach reached a satisfactory clearance within 3\u2009days in all neural tissues as well as several non\u2010neural tissues such as colon, duodenum, and pancreas. Compared to the CUBIC approach, all tissues reserved strong GFP fluorescence. Robust GFP fluorescence was visualized in sensory neuronal soma but not in sympathetic ganglia neuronal soma. On the other hand, GFP fluorescence in the TRPV1 cells appeared to be expressed throughout the epithelium of the duodenum and colon and the arteriole smooth muscle in all non\u2010neuronal tissues.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>This study shows that our combined PACT and CUBIC (CPC) protocol can clear tissues in significantly less time while preserving tissue integrity and fluorescence.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1111\/apha.14275","type":"journal-article","created":{"date-parts":[[2025,1,16]],"date-time":"2025-01-16T10:54:43Z","timestamp":1737024883000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Anatomical mapping of neural lineages expressing the transient receptor potential vanilloid type 1 receptor using a modified and combined\n                    <scp>PACT<\/scp>\n                    and\n                    <scp>CUBIC<\/scp>\n                    protocol for rapid tissue clearance"],"prefix":"10.1111","volume":"241","author":[{"given":"Oliver","family":"Kitzerow","sequence":"first","affiliation":[{"name":"Department of Genetics Cell Biology and Anatomy University of Nebraska Medical Center  Omaha Nebraska USA"},{"name":"Deptrtment of Anesthesiology University of Nebraska Medical Center  Omaha Nebraska USA"},{"name":"Department of Medical Education Creighton University  Omaha Nebraska USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samuel","family":"Christensen","sequence":"additional","affiliation":[{"name":"Deptrtment of Anesthesiology University of Nebraska Medical Center  Omaha Nebraska USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan","family":"Hong","sequence":"additional","affiliation":[{"name":"Deptrtment of Anesthesiology University of Nebraska Medical Center  Omaha Nebraska USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryan J.","family":"Adam","sequence":"additional","affiliation":[{"name":"Deptrtment of Anesthesiology University of Nebraska Medical Center  Omaha Nebraska USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Irving H.","family":"Zucker","sequence":"additional","affiliation":[{"name":"Department of Cellular and Integrative Physiology University of Nebraska Medical Center  Omaha Nebraska USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heather","family":"Jensen\u2010Smith","sequence":"additional","affiliation":[{"name":"Department of Genetics Cell Biology and Anatomy University of Nebraska Medical Center  Omaha Nebraska USA"},{"name":"Fred and Pamela Buffett Cancer Center University of Nebraska Medical Center  Omaha Nebraska 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