A Method to Visualize Cell Proliferation of Arabidopsis thaliana: A Case Study of the Root Apical Meristem
ABSTRACT Plant growth and development rely on a delicate balance between cell proliferation and cell differentiation. The root apical meristem (RAM) of Arabidopsis thaliana is an excellent model to study the cell cycle due to the coordinated relationship between nucleus shape and cell size at each s...
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| Format: | Article |
| Language: | English |
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Wiley
2025-03-01
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| Series: | Plant Direct |
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| Online Access: | https://doi.org/10.1002/pld3.70060 |
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| author | J. Irepan Reyes‐Olalde Miguel Tapia‐Rodríguez Vadim Pérez‐Koldenkova Gastón Contreras‐Jiménez Paul Hernández‐Herrera Gabriel Corkidi Arturo J. Arciniega‐González Maria De La Paz‐Sánchez Berenice García‐Ponce Adriana Garay‐Arroyo Elena R. Álvarez‐Buylla |
| author_facet | J. Irepan Reyes‐Olalde Miguel Tapia‐Rodríguez Vadim Pérez‐Koldenkova Gastón Contreras‐Jiménez Paul Hernández‐Herrera Gabriel Corkidi Arturo J. Arciniega‐González Maria De La Paz‐Sánchez Berenice García‐Ponce Adriana Garay‐Arroyo Elena R. Álvarez‐Buylla |
| author_sort | J. Irepan Reyes‐Olalde |
| collection | DOAJ |
| description | ABSTRACT Plant growth and development rely on a delicate balance between cell proliferation and cell differentiation. The root apical meristem (RAM) of Arabidopsis thaliana is an excellent model to study the cell cycle due to the coordinated relationship between nucleus shape and cell size at each stage, allowing for precise estimation of the cell cycle duration. In this study, we present a method for high‐resolution visualization of RAM cells. This is the first protocol that allows for simultaneous high‐resolution imaging of cellular and nuclear stains, being compatible with DNA replication markers such as EdU, including fluorescent proteins (H2B::YFP), SYTOX DNA stains, and the cell wall stain SR2200. This protocol includes a clarification procedure that enables the acquisition of high‐resolution 3D images, suitable for detailed subsequent analysis. |
| format | Article |
| id | doaj-art-5da26d207f724b56bb3de2b263531d87 |
| institution | OA Journals |
| issn | 2475-4455 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Wiley |
| record_format | Article |
| series | Plant Direct |
| spelling | doaj-art-5da26d207f724b56bb3de2b263531d872025-08-20T02:29:34ZengWileyPlant Direct2475-44552025-03-0194n/an/a10.1002/pld3.70060A Method to Visualize Cell Proliferation of Arabidopsis thaliana: A Case Study of the Root Apical MeristemJ. Irepan Reyes‐Olalde0Miguel Tapia‐Rodríguez1Vadim Pérez‐Koldenkova2Gastón Contreras‐Jiménez3Paul Hernández‐Herrera4Gabriel Corkidi5Arturo J. Arciniega‐González6Maria De La Paz‐Sánchez7Berenice García‐Ponce8Adriana Garay‐Arroyo9Elena R. Álvarez‐Buylla10Laboratorio de Genética Molecular, Epigenética, Desarrollo y Evolución de Plantas, Instituto de Ecología Universidad Nacional Autónoma de México, Ciudad Universitaria Ciudad de México MéxicoUnidad de Microscopía, Instituto de Investigaciones Biomédicas Universidad Nacional Autónoma de México, Ciudad Universitaria Ciudad de México MéxicoLaboratorio Nacional de Microscopía Avanzada, Centro Médico Nacional Siglo XXI‐IMSS Instituto Mexicano del Seguro Social Ciudad de México MexicoLaboratorio de Genética Molecular, Epigenética, Desarrollo y Evolución de Plantas, Instituto de Ecología Universidad Nacional Autónoma de México, Ciudad Universitaria Ciudad de México MéxicoDepartamento de Ingenería Celular y Biocatálisis, Laboratorio de Imágenes y Visión por Computadora Instituto de Biotecnología, UNAM Cuernavaca MéxicoDepartamento de Ingenería Celular y Biocatálisis, Laboratorio de Imágenes y Visión por Computadora Instituto de Biotecnología, UNAM Cuernavaca MéxicoLaboratorio de Genética Molecular, Epigenética, Desarrollo y Evolución de Plantas, Instituto de Ecología Universidad Nacional Autónoma de México, Ciudad Universitaria Ciudad de México MéxicoLaboratorio de Genética Molecular, Epigenética, Desarrollo y Evolución de Plantas, Instituto de Ecología Universidad Nacional Autónoma de México, Ciudad Universitaria Ciudad de México MéxicoLaboratorio de Genética Molecular, Epigenética, Desarrollo y Evolución de Plantas, Instituto de Ecología Universidad Nacional Autónoma de México, Ciudad Universitaria Ciudad de México MéxicoLaboratorio de Genética Molecular, Epigenética, Desarrollo y Evolución de Plantas, Instituto de Ecología Universidad Nacional Autónoma de México, Ciudad Universitaria Ciudad de México MéxicoLaboratorio de Genética Molecular, Epigenética, Desarrollo y Evolución de Plantas, Instituto de Ecología Universidad Nacional Autónoma de México, Ciudad Universitaria Ciudad de México MéxicoABSTRACT Plant growth and development rely on a delicate balance between cell proliferation and cell differentiation. The root apical meristem (RAM) of Arabidopsis thaliana is an excellent model to study the cell cycle due to the coordinated relationship between nucleus shape and cell size at each stage, allowing for precise estimation of the cell cycle duration. In this study, we present a method for high‐resolution visualization of RAM cells. This is the first protocol that allows for simultaneous high‐resolution imaging of cellular and nuclear stains, being compatible with DNA replication markers such as EdU, including fluorescent proteins (H2B::YFP), SYTOX DNA stains, and the cell wall stain SR2200. This protocol includes a clarification procedure that enables the acquisition of high‐resolution 3D images, suitable for detailed subsequent analysis.https://doi.org/10.1002/pld3.700605‐ethynyl‐2‐deoxyuridine (EdU)cell and nuclear markerscell cycleroot apical meristem |
| spellingShingle | J. Irepan Reyes‐Olalde Miguel Tapia‐Rodríguez Vadim Pérez‐Koldenkova Gastón Contreras‐Jiménez Paul Hernández‐Herrera Gabriel Corkidi Arturo J. Arciniega‐González Maria De La Paz‐Sánchez Berenice García‐Ponce Adriana Garay‐Arroyo Elena R. Álvarez‐Buylla A Method to Visualize Cell Proliferation of Arabidopsis thaliana: A Case Study of the Root Apical Meristem Plant Direct 5‐ethynyl‐2‐deoxyuridine (EdU) cell and nuclear markers cell cycle root apical meristem |
| title | A Method to Visualize Cell Proliferation of Arabidopsis thaliana: A Case Study of the Root Apical Meristem |
| title_full | A Method to Visualize Cell Proliferation of Arabidopsis thaliana: A Case Study of the Root Apical Meristem |
| title_fullStr | A Method to Visualize Cell Proliferation of Arabidopsis thaliana: A Case Study of the Root Apical Meristem |
| title_full_unstemmed | A Method to Visualize Cell Proliferation of Arabidopsis thaliana: A Case Study of the Root Apical Meristem |
| title_short | A Method to Visualize Cell Proliferation of Arabidopsis thaliana: A Case Study of the Root Apical Meristem |
| title_sort | method to visualize cell proliferation of arabidopsis thaliana a case study of the root apical meristem |
| topic | 5‐ethynyl‐2‐deoxyuridine (EdU) cell and nuclear markers cell cycle root apical meristem |
| url | https://doi.org/10.1002/pld3.70060 |
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