{"doi":"10.1002/cpz1.823","title":"Tissue‐Specific RNA Localization in the Uterus During Implantation, Decidualization and Placentation: Technical Nuances of Various Labeling Approaches","abstract":"<jats:title>Abstract</jats:title><jats:p><jats:italic>In situ</jats:italic> hybridization (ISH) is a sensitive method used to localize a specific sequence of DNA or RNA in biological samples, including cells, tissue sections or whole organs. RNA ISH can be used to determine spatial gene expression using a single‐stranded probe with a reverse‐complementary sequence. Cell‐specific gene expression has been studied using mRNA and protein levels. Signals produced by RNA probes are usually more specific than those produced by antibodies in immunostaining. Currently, ISH is the most widely used method to localize mRNA molecules. Traditionally, probes were labeled with radioactive isotopes, but the cumbersome procedures and potential health risk limit their acceptance. Recently, probes labeled with nonradioactive materials including digoxigenin, biotin and various fluorophores have been developed. The tyramide signal amplification system further enhances the sensitivity of detection. These methods have been applied in numerous studies in various tissues including reproductive organs. This article details three methods of RNA <jats:italic>in situ</jats:italic> hybridization: radioactive <jats:italic>in situ</jats:italic> hybridization, digoxigenin <jats:italic>in situ</jats:italic> hybridization, and digoxigenin‐tyramide signal amplification fluorescein <jats:italic>in situ</jats:italic> hybridization. The pros and cons of each protocol are discussed. © 2023 Wiley Periodicals LLC.</jats:p><jats:p><jats:bold>Basic Protocol 1</jats:bold>: Radioactive <jats:italic>in situ</jats:italic> hybridization (radioactive‐ISH)</jats:p><jats:p><jats:bold>Basic Protocol 2</jats:bold>: Digoxigenin <jats:italic>in situ</jats:italic> hybridization (DIG‐ISH)</jats:p><jats:p><jats:bold>Basic Protocol 3</jats:bold>: Digoxigenin‐tyramide signal amplification fluorescein <jats:italic>in situ</jats:italic> hybridization (DIG‐TSA‐FISH)</jats:p>","journal":"Current Protocols","year":2023,"id":41296,"datarank":0.3179840647900255,"base_score":1.6094379124341003,"endowment":1.6094379124341003,"self_citation_contribution":0.24141568686511508,"citation_network_contribution":0.07656837792491042,"self_endowment_contribution":0.24141568686511508,"citer_contribution":0.07656837792491042,"corpus_percentile":null,"corpus_rank":null,"citation_count":4,"citer_count":4,"citers_with_citation_signal":2,"citers_with_endowment":2,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":199128,"name":"Wenbo Deng","orcid":null,"position":1,"is_corresponding":false},{"id":199129,"name":"Xiaofei Sun","orcid":null,"position":2,"is_corresponding":false},{"id":199130,"name":"Sudhansu K. Dey","orcid":null,"position":3,"is_corresponding":false},{"id":199127,"name":"Amanda Dewar","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"base_score":1.6094379124341003,"endowment":1.6094379124341003,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"37345986","pmcid":"PMC10299818","openalex_id":"https://openalex.org/W4381597658","authors":[],"funders":[{"funder_name":"NICHD NIH HHS","grant_id":"R01 HD068524","title":null},{"funder_name":"NICHD NIH HHS","grant_id":"R01 HD103475","title":null}],"total_grants":2,"fwci":0.6233,"citation_percentile":0.71746959,"influential_citations":0,"citation_trend":[{"year":2024,"count":2},{"year":2026,"count":2}],"oa_status":"green","license":"http://onlinelibrary.wiley.com/termsAndConditions#vor","oa_locations":[{"url":"https://pmc.ncbi.nlm.nih.gov/articles/PMC10299818/pdf/nihms-1906369.pdf","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10299818","host_type":"GREEN"},{"url":"https://pmc.ncbi.nlm.nih.gov/articles/PMC10299818/pdf/nihms-1906369.pdf","host_type":"repository"},{"url":"https://currentprotocols.onlinelibrary.wiley.com/doi/pdf/10.1002/cpz1.823","host_type":"publisher"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/10299818","host_type":"repository"},{"url":"http://dx.doi.org/10.1002/cpz1.823","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/37345986","host_type":"repository"}],"fields_of_study":["Reproductive System and Pregnancy","RNA Research and Splicing","Reproductive Physiology in Livestock","Medicine","Biology","Female","Pregnancy","Humans","Digoxigenin","Placentation","RNA, Messenger","RNA","Uterus","Fluoresceins"],"mesh_terms":["Digoxigenin","Female","Fluoresceins","Humans","Placentation","Pregnancy","RNA","RNA, Messenger","Uterus"],"keywords":["Digoxigenin","In situ hybridization","Dig","Biology","Riboprobe","Molecular biology","Hybridization probe","RNA","Gene expression","Gene","Biochemistry","FISH","RNA probe","Radioactive","Tyramide Signal Amplification"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-06-12T23:11:45.076944Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}