Distinct effects of Fgf7 and Fgf10 on the terminal differentiation of murine bladder urothelium revealed using an organoid culture system
Abstract Background Dysregulation of the terminal differentiation of bladder urothelium is associated with the pathogenesis of urinary tract disorders. Fibroblast growth factor (Fgf)7 and Fgf10 stimulate urothelial proliferation; however, their roles in cellular differentiation remain unclear. In th...
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| Format: | Article |
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BMC
2023-10-01
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| Series: | BMC Urology |
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| Online Access: | https://doi.org/10.1186/s12894-023-01338-y |
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| author | Kazuto Suda Yuka Matsumoto Takanori Ochi Hiroyuki Koga Nobutaka Hattori Atsuyuki Yamataka Tetsuya Nakamura |
| author_facet | Kazuto Suda Yuka Matsumoto Takanori Ochi Hiroyuki Koga Nobutaka Hattori Atsuyuki Yamataka Tetsuya Nakamura |
| author_sort | Kazuto Suda |
| collection | DOAJ |
| description | Abstract Background Dysregulation of the terminal differentiation of bladder urothelium is associated with the pathogenesis of urinary tract disorders. Fibroblast growth factor (Fgf)7 and Fgf10 stimulate urothelial proliferation; however, their roles in cellular differentiation remain unclear. In this study, we used an organoid system to investigate the roles of these Fgfs in regulating bladder urothelium differentiation and identify their distribution patterns in the mouse bladder. Methods Adult bladder epithelia (AdBE) isolated from adult mouse bladder tissues (AdBTs) were used to culture adult bladder organoids (AdBOs) in the presence of Fgf7 and Fgf10. The differentiation status of the cells in AdBTs, AdBEs, AdBOs, and neonatal bladder tissues (NeoBTs) was analyzed via quantitative real-time-PCR for the presence of undifferentiated cell markers (Krt5, Trp63, and Krt14) and differentiated cell markers (Krt20, Upk1a, Upk2, and Upk3a). Organoid cell proliferation was assessed by counting cell numbers using the trypan blue method. The effects of Fgf7 and Fgf10 on organoid differentiation were assessed using different doses of Fgfs, and the involvement of peroxisome proliferator-activated receptor γ (PPARγ) signaling in these processes was tested by introducing a PPARγ agonist (Rosiglitazone) and antagonist (T0070907) to the culture. The expression patterns of Fgf7 and Fgf10 were examined via in situ hybridization of AdBTs. Results AdBOs showed higher expression of undifferentiated cell markers and lower expression of differentiated cell markers than AdBTs, NeoBTs, and AdBEs, indicating the relatively immature state of AdBOs. Differentiation of AdBOs was enhanced by Rosiglitazone and Fgf7, suggesting an interplay of intracellular signals between Fgf7 and PPARγ. Co-addition of T0070907 suppressed Fgf7-mediated differentiation, demonstrating that PPARγ is activated downstream of Fgf7 to promote cellular differentiation into umbrella cells. Furthermore, we found that Fgf7 is predominantly expressed in the umbrella cells of the urothelium, whereas Fgf10 is predominantly expressed in the urothelium and stroma of AdBTs. Conclusions We demonstrated that unlike Fgf10, Fgf7 induces cellular differentiation via PPARγ activity and has a unique tissue distribution pattern in the adult bladder. Further studies on the Fgf7-PPARγ signaling axis would provide insights into the differentiation mechanisms toward functional umbrella cells and the pathogenesis of several urinary tract diseases. |
| first_indexed | 2024-03-10T16:57:34Z |
| format | Article |
| id | doaj.art-18596117a06c4f3d8e2d88f2ab7b55a1 |
| institution | Directory Open Access Journal |
| issn | 1471-2490 |
| language | English |
| last_indexed | 2024-03-10T16:57:34Z |
| publishDate | 2023-10-01 |
| publisher | BMC |
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| series | BMC Urology |
| spelling | doaj.art-18596117a06c4f3d8e2d88f2ab7b55a12023-11-20T11:03:31ZengBMCBMC Urology1471-24902023-10-0123111410.1186/s12894-023-01338-yDistinct effects of Fgf7 and Fgf10 on the terminal differentiation of murine bladder urothelium revealed using an organoid culture systemKazuto Suda0Yuka Matsumoto1Takanori Ochi2Hiroyuki Koga3Nobutaka Hattori4Atsuyuki Yamataka5Tetsuya Nakamura6Department of Pediatric General and Urogenital Surgery, Juntendo University School of MedicineDepartment of Pediatric General and Urogenital Surgery, Juntendo University School of MedicineDepartment of Pediatric General and Urogenital Surgery, Juntendo University School of MedicineDepartment of Pediatric General and Urogenital Surgery, Juntendo University School of MedicineDepartment of Neurology, Juntendo University School of MedicineDepartment of Pediatric General and Urogenital Surgery, Juntendo University School of MedicineDepartment of Research and Development for Organoids, Juntendo University Graduate School of MedicineAbstract Background Dysregulation of the terminal differentiation of bladder urothelium is associated with the pathogenesis of urinary tract disorders. Fibroblast growth factor (Fgf)7 and Fgf10 stimulate urothelial proliferation; however, their roles in cellular differentiation remain unclear. In this study, we used an organoid system to investigate the roles of these Fgfs in regulating bladder urothelium differentiation and identify their distribution patterns in the mouse bladder. Methods Adult bladder epithelia (AdBE) isolated from adult mouse bladder tissues (AdBTs) were used to culture adult bladder organoids (AdBOs) in the presence of Fgf7 and Fgf10. The differentiation status of the cells in AdBTs, AdBEs, AdBOs, and neonatal bladder tissues (NeoBTs) was analyzed via quantitative real-time-PCR for the presence of undifferentiated cell markers (Krt5, Trp63, and Krt14) and differentiated cell markers (Krt20, Upk1a, Upk2, and Upk3a). Organoid cell proliferation was assessed by counting cell numbers using the trypan blue method. The effects of Fgf7 and Fgf10 on organoid differentiation were assessed using different doses of Fgfs, and the involvement of peroxisome proliferator-activated receptor γ (PPARγ) signaling in these processes was tested by introducing a PPARγ agonist (Rosiglitazone) and antagonist (T0070907) to the culture. The expression patterns of Fgf7 and Fgf10 were examined via in situ hybridization of AdBTs. Results AdBOs showed higher expression of undifferentiated cell markers and lower expression of differentiated cell markers than AdBTs, NeoBTs, and AdBEs, indicating the relatively immature state of AdBOs. Differentiation of AdBOs was enhanced by Rosiglitazone and Fgf7, suggesting an interplay of intracellular signals between Fgf7 and PPARγ. Co-addition of T0070907 suppressed Fgf7-mediated differentiation, demonstrating that PPARγ is activated downstream of Fgf7 to promote cellular differentiation into umbrella cells. Furthermore, we found that Fgf7 is predominantly expressed in the umbrella cells of the urothelium, whereas Fgf10 is predominantly expressed in the urothelium and stroma of AdBTs. Conclusions We demonstrated that unlike Fgf10, Fgf7 induces cellular differentiation via PPARγ activity and has a unique tissue distribution pattern in the adult bladder. Further studies on the Fgf7-PPARγ signaling axis would provide insights into the differentiation mechanisms toward functional umbrella cells and the pathogenesis of several urinary tract diseases.https://doi.org/10.1186/s12894-023-01338-yBladderUrotheliumDifferentiationOrganoidFibroblast growth factor 7Fibroblast growth factor 10 |
| spellingShingle | Kazuto Suda Yuka Matsumoto Takanori Ochi Hiroyuki Koga Nobutaka Hattori Atsuyuki Yamataka Tetsuya Nakamura Distinct effects of Fgf7 and Fgf10 on the terminal differentiation of murine bladder urothelium revealed using an organoid culture system BMC Urology Bladder Urothelium Differentiation Organoid Fibroblast growth factor 7 Fibroblast growth factor 10 |
| title | Distinct effects of Fgf7 and Fgf10 on the terminal differentiation of murine bladder urothelium revealed using an organoid culture system |
| title_full | Distinct effects of Fgf7 and Fgf10 on the terminal differentiation of murine bladder urothelium revealed using an organoid culture system |
| title_fullStr | Distinct effects of Fgf7 and Fgf10 on the terminal differentiation of murine bladder urothelium revealed using an organoid culture system |
| title_full_unstemmed | Distinct effects of Fgf7 and Fgf10 on the terminal differentiation of murine bladder urothelium revealed using an organoid culture system |
| title_short | Distinct effects of Fgf7 and Fgf10 on the terminal differentiation of murine bladder urothelium revealed using an organoid culture system |
| title_sort | distinct effects of fgf7 and fgf10 on the terminal differentiation of murine bladder urothelium revealed using an organoid culture system |
| topic | Bladder Urothelium Differentiation Organoid Fibroblast growth factor 7 Fibroblast growth factor 10 |
| url | https://doi.org/10.1186/s12894-023-01338-y |
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