Intronic RNAscope probes enable precise identification of cardiomyocyte nuclei and cell cycle activity
Abstract Cardiac regeneration studies have been plagued by technical challenges in unequivocally identifying cardiomyocyte (CM) nuclei in cardiac sections, crucial for accurate identification of cycling CMs. The use of antibodies to sarcomeric proteins is error-prone, the CM specificity of common nu...
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Nature Portfolio
2025-04-01
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| Series: | Communications Biology |
| Online Access: | https://doi.org/10.1038/s42003-025-08012-z |
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| author | Zhe Yu Sen Zhang Julius Bogomolovas Ju Chen Sylvia M. Evans |
| author_facet | Zhe Yu Sen Zhang Julius Bogomolovas Ju Chen Sylvia M. Evans |
| author_sort | Zhe Yu |
| collection | DOAJ |
| description | Abstract Cardiac regeneration studies have been plagued by technical challenges in unequivocally identifying cardiomyocyte (CM) nuclei in cardiac sections, crucial for accurate identification of cycling CMs. The use of antibodies to sarcomeric proteins is error-prone, the CM specificity of common nuclear markers is controversial, and utilizing genetically modified mouse models poses risk of inducing unintended cardiac phenotypes. The application of RNAscope intronic probes overcomes the above shortcomings. Intronic probes label intronic RNAs within nuclei and can therefore be utilized as a method for nuclear localization. A Tnnt2 intronic RNAscope probe highly colocalized with Obscurin-H2B-GFP in adult mouse hearts, demonstrating CM specificity. Studies in embryos demonstrated that the Tnnt2 intronic RNAscope probe labeled CM nuclei that had undergone DNA replication, and remained closely associated with CM chromatin at all stages of mitosis, even with nuclear envelope breakdown. The efficiency, accuracy, and perdurance of the Tnnt2 intronic RNAscope probe even with nuclear envelope breakdown facilitated reliable investigation of dynamics of DNA synthesis and potential mitoses in CMs in both border and infarct zones after myocardial infarction (MI). Furthermore, we designed Myl2 and Myl4 intronic RNAscope probes, which labeled ventricular and atrial CM nuclei, respectively, and may help identify CM subtypes generated in vitro. |
| format | Article |
| id | doaj-art-754b716e9cd643e3b518120d5a2cc85a |
| institution | OA Journals |
| issn | 2399-3642 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Nature Portfolio |
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| series | Communications Biology |
| spelling | doaj-art-754b716e9cd643e3b518120d5a2cc85a2025-08-20T02:11:47ZengNature PortfolioCommunications Biology2399-36422025-04-018111110.1038/s42003-025-08012-zIntronic RNAscope probes enable precise identification of cardiomyocyte nuclei and cell cycle activityZhe Yu0Sen Zhang1Julius Bogomolovas2Ju Chen3Sylvia M. Evans4Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San DiegoDepartment of Pharmacology & Regenerative Medicine, University of Illinois ChicagoDepartment of Medicine, University of California San DiegoDepartment of Medicine, University of California San DiegoSkaggs School of Pharmacy and Pharmaceutical Sciences, University of California San DiegoAbstract Cardiac regeneration studies have been plagued by technical challenges in unequivocally identifying cardiomyocyte (CM) nuclei in cardiac sections, crucial for accurate identification of cycling CMs. The use of antibodies to sarcomeric proteins is error-prone, the CM specificity of common nuclear markers is controversial, and utilizing genetically modified mouse models poses risk of inducing unintended cardiac phenotypes. The application of RNAscope intronic probes overcomes the above shortcomings. Intronic probes label intronic RNAs within nuclei and can therefore be utilized as a method for nuclear localization. A Tnnt2 intronic RNAscope probe highly colocalized with Obscurin-H2B-GFP in adult mouse hearts, demonstrating CM specificity. Studies in embryos demonstrated that the Tnnt2 intronic RNAscope probe labeled CM nuclei that had undergone DNA replication, and remained closely associated with CM chromatin at all stages of mitosis, even with nuclear envelope breakdown. The efficiency, accuracy, and perdurance of the Tnnt2 intronic RNAscope probe even with nuclear envelope breakdown facilitated reliable investigation of dynamics of DNA synthesis and potential mitoses in CMs in both border and infarct zones after myocardial infarction (MI). Furthermore, we designed Myl2 and Myl4 intronic RNAscope probes, which labeled ventricular and atrial CM nuclei, respectively, and may help identify CM subtypes generated in vitro.https://doi.org/10.1038/s42003-025-08012-z |
| spellingShingle | Zhe Yu Sen Zhang Julius Bogomolovas Ju Chen Sylvia M. Evans Intronic RNAscope probes enable precise identification of cardiomyocyte nuclei and cell cycle activity Communications Biology |
| title | Intronic RNAscope probes enable precise identification of cardiomyocyte nuclei and cell cycle activity |
| title_full | Intronic RNAscope probes enable precise identification of cardiomyocyte nuclei and cell cycle activity |
| title_fullStr | Intronic RNAscope probes enable precise identification of cardiomyocyte nuclei and cell cycle activity |
| title_full_unstemmed | Intronic RNAscope probes enable precise identification of cardiomyocyte nuclei and cell cycle activity |
| title_short | Intronic RNAscope probes enable precise identification of cardiomyocyte nuclei and cell cycle activity |
| title_sort | intronic rnascope probes enable precise identification of cardiomyocyte nuclei and cell cycle activity |
| url | https://doi.org/10.1038/s42003-025-08012-z |
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