Small Molecules Temporarily Induce Neuronal Features in Adult Canine Dermal Fibroblasts
Several methods have been developed to generate neurons from other cell types for performing regeneration therapy and in vitro studies of central nerve disease. Small molecules (SMs) can efficiently induce neuronal features in human and rodent fibroblasts without transgenes. Although canines have be...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-10-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/21/15804 |
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| author | Kiyotaka Arai Fumiyo Saito Masashi Miyazaki Haruto Kushige Yayoi Izu Noritaka Maeta Kazuaki Yamazoe |
| author_facet | Kiyotaka Arai Fumiyo Saito Masashi Miyazaki Haruto Kushige Yayoi Izu Noritaka Maeta Kazuaki Yamazoe |
| author_sort | Kiyotaka Arai |
| collection | DOAJ |
| description | Several methods have been developed to generate neurons from other cell types for performing regeneration therapy and in vitro studies of central nerve disease. Small molecules (SMs) can efficiently induce neuronal features in human and rodent fibroblasts without transgenes. Although canines have been used as a spontaneous disease model of human central nerve, efficient neuronal reprogramming method of canine cells have not been well established. We aimed to induce neuronal features in adult canine dermal fibroblasts (ACDFs) by SMs and assess the permanency of these changes. ACDFs treated with eight SMs developed a round-shaped cell body with branching processes and expressed neuronal proteins, including βIII-tubulin, microtubule-associated protein 2 (MAP2), and neurofilament-medium. Transcriptome profiling revealed the upregulation of neuron-related genes, such as <i>SNAP25</i> and <i>GRIA4</i>, and downregulation of fibroblast-related genes, such as <i>COL12A1</i> and <i>CCN5</i>. Calcium fluorescent imaging demonstrated an increase in intracellular Ca<sup>2+</sup> concentration upon stimulation with glutamate and KCl. Although neuronal features were induced similarly in basement membrane extract droplet culture, they diminished after culturing without SMs or in vivo transplantation into an injured spinal cord. In conclusion, SMs temporarily induce neuronal features in ACDFs. However, the analysis of bottlenecks in the neuronal induction is crucial for optimizing the process. |
| first_indexed | 2024-03-11T11:28:41Z |
| format | Article |
| id | doaj.art-69584671da2f4ce0a5f07676cd4418ad |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T11:28:41Z |
| publishDate | 2023-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-69584671da2f4ce0a5f07676cd4418ad2023-11-10T15:05:23ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-10-0124211580410.3390/ijms242115804Small Molecules Temporarily Induce Neuronal Features in Adult Canine Dermal FibroblastsKiyotaka Arai0Fumiyo Saito1Masashi Miyazaki2Haruto Kushige3Yayoi Izu4Noritaka Maeta5Kazuaki Yamazoe6Department of Veterinary Surgery, Faculty of Veterinary Medicine, Okayama University of Science, 1-3 Ikoi-no-oka, Imabari 794-8555, JapanDepartment of Toxicology, Faculty of Veterinary Medicine, Okayama University of Science, 1-3 Ikoi-no-oka, Imabari 794-8555, JapanDepartment of Veterinary Surgery, Faculty of Veterinary Medicine, Okayama University of Science, 1-3 Ikoi-no-oka, Imabari 794-8555, JapanDepartment of Veterinary Surgery, Faculty of Veterinary Medicine, Okayama University of Science, 1-3 Ikoi-no-oka, Imabari 794-8555, JapanDepartment of Laboratory Animal Science, Faculty of Veterinary Medicine, Okayama University of Science, 1-3 Ikoi-no-oka, Imabari 794-8555, JapanDepartment of Veterinary Surgery, Faculty of Veterinary Medicine, Okayama University of Science, 1-3 Ikoi-no-oka, Imabari 794-8555, JapanDepartment of Veterinary Surgery, Faculty of Veterinary Medicine, Okayama University of Science, 1-3 Ikoi-no-oka, Imabari 794-8555, JapanSeveral methods have been developed to generate neurons from other cell types for performing regeneration therapy and in vitro studies of central nerve disease. Small molecules (SMs) can efficiently induce neuronal features in human and rodent fibroblasts without transgenes. Although canines have been used as a spontaneous disease model of human central nerve, efficient neuronal reprogramming method of canine cells have not been well established. We aimed to induce neuronal features in adult canine dermal fibroblasts (ACDFs) by SMs and assess the permanency of these changes. ACDFs treated with eight SMs developed a round-shaped cell body with branching processes and expressed neuronal proteins, including βIII-tubulin, microtubule-associated protein 2 (MAP2), and neurofilament-medium. Transcriptome profiling revealed the upregulation of neuron-related genes, such as <i>SNAP25</i> and <i>GRIA4</i>, and downregulation of fibroblast-related genes, such as <i>COL12A1</i> and <i>CCN5</i>. Calcium fluorescent imaging demonstrated an increase in intracellular Ca<sup>2+</sup> concentration upon stimulation with glutamate and KCl. Although neuronal features were induced similarly in basement membrane extract droplet culture, they diminished after culturing without SMs or in vivo transplantation into an injured spinal cord. In conclusion, SMs temporarily induce neuronal features in ACDFs. However, the analysis of bottlenecks in the neuronal induction is crucial for optimizing the process.https://www.mdpi.com/1422-0067/24/21/15804small moleculesadult canine dermal fibroblastsneuronal featuresneuronal induction |
| spellingShingle | Kiyotaka Arai Fumiyo Saito Masashi Miyazaki Haruto Kushige Yayoi Izu Noritaka Maeta Kazuaki Yamazoe Small Molecules Temporarily Induce Neuronal Features in Adult Canine Dermal Fibroblasts International Journal of Molecular Sciences small molecules adult canine dermal fibroblasts neuronal features neuronal induction |
| title | Small Molecules Temporarily Induce Neuronal Features in Adult Canine Dermal Fibroblasts |
| title_full | Small Molecules Temporarily Induce Neuronal Features in Adult Canine Dermal Fibroblasts |
| title_fullStr | Small Molecules Temporarily Induce Neuronal Features in Adult Canine Dermal Fibroblasts |
| title_full_unstemmed | Small Molecules Temporarily Induce Neuronal Features in Adult Canine Dermal Fibroblasts |
| title_short | Small Molecules Temporarily Induce Neuronal Features in Adult Canine Dermal Fibroblasts |
| title_sort | small molecules temporarily induce neuronal features in adult canine dermal fibroblasts |
| topic | small molecules adult canine dermal fibroblasts neuronal features neuronal induction |
| url | https://www.mdpi.com/1422-0067/24/21/15804 |
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