Active Phagocytosis and Diachronic Processing of Calcium Oxalate Monohydrate Crystals in an in vitro Macrophage Model
Background: We previously discovered that renal macrophages (Mφs) phagocytose renal calcium oxalate monohydrate (COM) crystals. This study investigated the processing of engulfed crystals using in vitro models. Methods: J774.1 mouse Mφs were exposed to COM crystals and observed for 24 h using polari...
| Main Authors: | , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Karger Publishers
2019-09-01
|
| Series: | Kidney & Blood Pressure Research |
| Subjects: | |
| Online Access: | https://www.karger.com/Article/FullText/501965 |
| _version_ | 1819205686133784576 |
|---|---|
| author | Atsushi Okada Hiromasa Aoki Daichi Onozato Taiki Kato Tadahiro Hashita Hiroshi Takase Teruaki Sugino Rei Unno Kazumi Taguchi Shuzo Hamamoto Ryosuke Ando Kentaro Mizuno Keiichi Tozawa Tamihide Matsunaga Kenjiro Kohri Takahiro Yasui |
| author_facet | Atsushi Okada Hiromasa Aoki Daichi Onozato Taiki Kato Tadahiro Hashita Hiroshi Takase Teruaki Sugino Rei Unno Kazumi Taguchi Shuzo Hamamoto Ryosuke Ando Kentaro Mizuno Keiichi Tozawa Tamihide Matsunaga Kenjiro Kohri Takahiro Yasui |
| author_sort | Atsushi Okada |
| collection | DOAJ |
| description | Background: We previously discovered that renal macrophages (Mφs) phagocytose renal calcium oxalate monohydrate (COM) crystals. This study investigated the processing of engulfed crystals using in vitro models. Methods: J774.1 mouse Mφs were exposed to COM crystals and observed for 24 h using polarized light microscopy with/without cytochalasin B (CB), an inhibitor of phagocytosis, to confirm active crystal phagocytosis. LysoTracker and immunohistochemical staining using transmission electron microscopy for lysosomal-associated membrane protein 1 were used to confirm engulfed COM crystal uptake into lysosomes. Diachronic tracking of specific Mφs was performed to capture the entire course of engulfed COM crystal processing using polarized light microscopy. Follow-up studies of fluorescent COM (f-COM) crystals using imaging cytometry were performed in the presence and absence of nigericin to dissipate the pH gradient in acidic organelles. Results: Phagocytosis rates increased with COM density and were significantly lower in cells treated with CB (p < 0.01). We observed that engulfed crystals colocalized within lysosomes of the Mφs; moreover, diachronic observation indicated that the engulfed COM crystals were subdivided during Mφ division and eliminated by the 7th day of culture. Additionally, imaging cytometry showed that the fluorescence level of f-COM crystals in the nigericin (–) group after 48 h was significantly lower than that in the nigericin (+) group. Conclusions: This study confirmed active phagocytosis and lysosomal processing of engulfed COM crystals by Mφs. This discovery is expected to contribute to the development of future drugs that enhance the COM crystal phagocytic ability of Mφs. |
| first_indexed | 2024-12-23T04:55:39Z |
| format | Article |
| id | doaj.art-6ada40212bbf4039bfbdc2e5015a900d |
| institution | Directory Open Access Journal |
| issn | 1420-4096 1423-0143 |
| language | English |
| last_indexed | 2024-12-23T04:55:39Z |
| publishDate | 2019-09-01 |
| publisher | Karger Publishers |
| record_format | Article |
| series | Kidney & Blood Pressure Research |
| spelling | doaj.art-6ada40212bbf4039bfbdc2e5015a900d2022-12-21T17:59:22ZengKarger PublishersKidney & Blood Pressure Research1420-40961423-01432019-09-0111210.1159/000501965501965Active Phagocytosis and Diachronic Processing of Calcium Oxalate Monohydrate Crystals in an in vitro Macrophage ModelAtsushi OkadaHiromasa AokiDaichi OnozatoTaiki KatoTadahiro HashitaHiroshi TakaseTeruaki SuginoRei UnnoKazumi TaguchiShuzo HamamotoRyosuke AndoKentaro MizunoKeiichi TozawaTamihide MatsunagaKenjiro KohriTakahiro YasuiBackground: We previously discovered that renal macrophages (Mφs) phagocytose renal calcium oxalate monohydrate (COM) crystals. This study investigated the processing of engulfed crystals using in vitro models. Methods: J774.1 mouse Mφs were exposed to COM crystals and observed for 24 h using polarized light microscopy with/without cytochalasin B (CB), an inhibitor of phagocytosis, to confirm active crystal phagocytosis. LysoTracker and immunohistochemical staining using transmission electron microscopy for lysosomal-associated membrane protein 1 were used to confirm engulfed COM crystal uptake into lysosomes. Diachronic tracking of specific Mφs was performed to capture the entire course of engulfed COM crystal processing using polarized light microscopy. Follow-up studies of fluorescent COM (f-COM) crystals using imaging cytometry were performed in the presence and absence of nigericin to dissipate the pH gradient in acidic organelles. Results: Phagocytosis rates increased with COM density and were significantly lower in cells treated with CB (p < 0.01). We observed that engulfed crystals colocalized within lysosomes of the Mφs; moreover, diachronic observation indicated that the engulfed COM crystals were subdivided during Mφ division and eliminated by the 7th day of culture. Additionally, imaging cytometry showed that the fluorescence level of f-COM crystals in the nigericin (–) group after 48 h was significantly lower than that in the nigericin (+) group. Conclusions: This study confirmed active phagocytosis and lysosomal processing of engulfed COM crystals by Mφs. This discovery is expected to contribute to the development of future drugs that enhance the COM crystal phagocytic ability of Mφs.https://www.karger.com/Article/FullText/501965Calcium oxalateKidneyMacrophagesNephrolithiasisPhagocytosis |
| spellingShingle | Atsushi Okada Hiromasa Aoki Daichi Onozato Taiki Kato Tadahiro Hashita Hiroshi Takase Teruaki Sugino Rei Unno Kazumi Taguchi Shuzo Hamamoto Ryosuke Ando Kentaro Mizuno Keiichi Tozawa Tamihide Matsunaga Kenjiro Kohri Takahiro Yasui Active Phagocytosis and Diachronic Processing of Calcium Oxalate Monohydrate Crystals in an in vitro Macrophage Model Kidney & Blood Pressure Research Calcium oxalate Kidney Macrophages Nephrolithiasis Phagocytosis |
| title | Active Phagocytosis and Diachronic Processing of Calcium Oxalate Monohydrate Crystals in an in vitro Macrophage Model |
| title_full | Active Phagocytosis and Diachronic Processing of Calcium Oxalate Monohydrate Crystals in an in vitro Macrophage Model |
| title_fullStr | Active Phagocytosis and Diachronic Processing of Calcium Oxalate Monohydrate Crystals in an in vitro Macrophage Model |
| title_full_unstemmed | Active Phagocytosis and Diachronic Processing of Calcium Oxalate Monohydrate Crystals in an in vitro Macrophage Model |
| title_short | Active Phagocytosis and Diachronic Processing of Calcium Oxalate Monohydrate Crystals in an in vitro Macrophage Model |
| title_sort | active phagocytosis and diachronic processing of calcium oxalate monohydrate crystals in an in vitro macrophage model |
| topic | Calcium oxalate Kidney Macrophages Nephrolithiasis Phagocytosis |
| url | https://www.karger.com/Article/FullText/501965 |
| work_keys_str_mv | AT atsushiokada activephagocytosisanddiachronicprocessingofcalciumoxalatemonohydratecrystalsinaninvitromacrophagemodel AT hiromasaaoki activephagocytosisanddiachronicprocessingofcalciumoxalatemonohydratecrystalsinaninvitromacrophagemodel AT daichionozato activephagocytosisanddiachronicprocessingofcalciumoxalatemonohydratecrystalsinaninvitromacrophagemodel AT taikikato activephagocytosisanddiachronicprocessingofcalciumoxalatemonohydratecrystalsinaninvitromacrophagemodel AT tadahirohashita activephagocytosisanddiachronicprocessingofcalciumoxalatemonohydratecrystalsinaninvitromacrophagemodel AT hiroshitakase activephagocytosisanddiachronicprocessingofcalciumoxalatemonohydratecrystalsinaninvitromacrophagemodel AT teruakisugino activephagocytosisanddiachronicprocessingofcalciumoxalatemonohydratecrystalsinaninvitromacrophagemodel AT reiunno activephagocytosisanddiachronicprocessingofcalciumoxalatemonohydratecrystalsinaninvitromacrophagemodel AT kazumitaguchi activephagocytosisanddiachronicprocessingofcalciumoxalatemonohydratecrystalsinaninvitromacrophagemodel AT shuzohamamoto activephagocytosisanddiachronicprocessingofcalciumoxalatemonohydratecrystalsinaninvitromacrophagemodel AT ryosukeando activephagocytosisanddiachronicprocessingofcalciumoxalatemonohydratecrystalsinaninvitromacrophagemodel AT kentaromizuno activephagocytosisanddiachronicprocessingofcalciumoxalatemonohydratecrystalsinaninvitromacrophagemodel AT keiichitozawa activephagocytosisanddiachronicprocessingofcalciumoxalatemonohydratecrystalsinaninvitromacrophagemodel AT tamihidematsunaga activephagocytosisanddiachronicprocessingofcalciumoxalatemonohydratecrystalsinaninvitromacrophagemodel AT kenjirokohri activephagocytosisanddiachronicprocessingofcalciumoxalatemonohydratecrystalsinaninvitromacrophagemodel AT takahiroyasui activephagocytosisanddiachronicprocessingofcalciumoxalatemonohydratecrystalsinaninvitromacrophagemodel |