Chloroquine inhibits artificial oocyte activation induced by ethanol or Sr2+ but not by sperm in mice
Calcium release from the endoplasmic reticulum via sperm-derived phospholipase C zeta is crucial for oocyte activation during fertilization. Chloroquine (CQ) inhibits the increase in cytoplasmic calcium. This study investigated the effects of CQ on fertilization and oocyte activation. Oocytes were c...
Saved in:
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
The Society for Reproduction and Development
2024-12-01
|
| Series: | The Journal of Reproduction and Development |
| Subjects: | |
| Online Access: | https://www.jstage.jst.go.jp/article/jrd/71/1/71_2024-089/_pdf/-char/en |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832540552569028608 |
|---|---|
| author | Tadashi YAMAZAKI Md Wasim BARI Satoshi KISHIGAMI |
| author_facet | Tadashi YAMAZAKI Md Wasim BARI Satoshi KISHIGAMI |
| author_sort | Tadashi YAMAZAKI |
| collection | DOAJ |
| description | Calcium release from the endoplasmic reticulum via sperm-derived phospholipase C zeta is crucial for oocyte activation during fertilization. Chloroquine (CQ) inhibits the increase in cytoplasmic calcium. This study investigated the effects of CQ on fertilization and oocyte activation. Oocytes were collected from ICR mice, and in vitro fertilization and artificial oocyte activation with strontium ions (Sr2+) and ethanol were performed in the presence of 50 µM CQ. Pronuclear formation was assessed via Hoechst33242 nuclear staining, cortical granule release was evaluated using lens culinaris agglutinin-fluorescein isothiocyanate staining, and cytosolic calcium levels were measured using fluorescence microscopy with Cal-520 AM. In the presence of CQ, no pronuclei were formed even 8 h after Sr2+-induced oocyte activation. Furthermore, cortical granule release in CQ-treated oocytes was significantly suppressed, although not completely inhibited, and no increase in cytosolic calcium was detected. CQ also inhibited pronuclear formation during ethanol-induced oocyte activation. In in vitro fertilization, although the fertilization rate was decreased in the CQ-treated group, in which CQ treatment was continuously applied during insemination, pronuclear formation and cortical granule release were observed. The decrease in the fertilization rate was likely attributable to reduced sperm motility and decreased penetration of the zona pellucida. The findings indicate that the oocyte activation pathways triggered by ethanol/Sr2+ and sperm are distinguishable by CQ, and that CQ can be used as a selective inhibitor of oocyte activation induced by Sr2+ or ethanol treatment. |
| format | Article |
| id | doaj-art-b2d65d053a754be5ae139b39a6ccf805 |
| institution | Kabale University |
| issn | 0916-8818 1348-4400 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | The Society for Reproduction and Development |
| record_format | Article |
| series | The Journal of Reproduction and Development |
| spelling | doaj-art-b2d65d053a754be5ae139b39a6ccf8052025-02-04T23:41:35ZengThe Society for Reproduction and DevelopmentThe Journal of Reproduction and Development0916-88181348-44002024-12-01711495410.1262/jrd.2024-089jrdChloroquine inhibits artificial oocyte activation induced by ethanol or Sr2+ but not by sperm in miceTadashi YAMAZAKI0Md Wasim BARI1Satoshi KISHIGAMI2Department of Integrated Applied Life Science, Integrated Graduate School of Medicine, Engineering, and Agricultural Sciences, University of Yamanashi, Yamanashi 400-8510, JapanFaculty of Life and Environmental Sciences, University of Yamanashi, Yamanashi 400-8510, JapanDepartment of Integrated Applied Life Science, Integrated Graduate School of Medicine, Engineering, and Agricultural Sciences, University of Yamanashi, Yamanashi 400-8510, JapanCalcium release from the endoplasmic reticulum via sperm-derived phospholipase C zeta is crucial for oocyte activation during fertilization. Chloroquine (CQ) inhibits the increase in cytoplasmic calcium. This study investigated the effects of CQ on fertilization and oocyte activation. Oocytes were collected from ICR mice, and in vitro fertilization and artificial oocyte activation with strontium ions (Sr2+) and ethanol were performed in the presence of 50 µM CQ. Pronuclear formation was assessed via Hoechst33242 nuclear staining, cortical granule release was evaluated using lens culinaris agglutinin-fluorescein isothiocyanate staining, and cytosolic calcium levels were measured using fluorescence microscopy with Cal-520 AM. In the presence of CQ, no pronuclei were formed even 8 h after Sr2+-induced oocyte activation. Furthermore, cortical granule release in CQ-treated oocytes was significantly suppressed, although not completely inhibited, and no increase in cytosolic calcium was detected. CQ also inhibited pronuclear formation during ethanol-induced oocyte activation. In in vitro fertilization, although the fertilization rate was decreased in the CQ-treated group, in which CQ treatment was continuously applied during insemination, pronuclear formation and cortical granule release were observed. The decrease in the fertilization rate was likely attributable to reduced sperm motility and decreased penetration of the zona pellucida. The findings indicate that the oocyte activation pathways triggered by ethanol/Sr2+ and sperm are distinguishable by CQ, and that CQ can be used as a selective inhibitor of oocyte activation induced by Sr2+ or ethanol treatment.https://www.jstage.jst.go.jp/article/jrd/71/1/71_2024-089/_pdf/-char/enartificial oocyte activationchloroquinein vitro fertilization (ivf) |
| spellingShingle | Tadashi YAMAZAKI Md Wasim BARI Satoshi KISHIGAMI Chloroquine inhibits artificial oocyte activation induced by ethanol or Sr2+ but not by sperm in mice The Journal of Reproduction and Development artificial oocyte activation chloroquine in vitro fertilization (ivf) |
| title | Chloroquine inhibits artificial oocyte activation induced by ethanol or Sr2+ but not by sperm in mice |
| title_full | Chloroquine inhibits artificial oocyte activation induced by ethanol or Sr2+ but not by sperm in mice |
| title_fullStr | Chloroquine inhibits artificial oocyte activation induced by ethanol or Sr2+ but not by sperm in mice |
| title_full_unstemmed | Chloroquine inhibits artificial oocyte activation induced by ethanol or Sr2+ but not by sperm in mice |
| title_short | Chloroquine inhibits artificial oocyte activation induced by ethanol or Sr2+ but not by sperm in mice |
| title_sort | chloroquine inhibits artificial oocyte activation induced by ethanol or sr2 but not by sperm in mice |
| topic | artificial oocyte activation chloroquine in vitro fertilization (ivf) |
| url | https://www.jstage.jst.go.jp/article/jrd/71/1/71_2024-089/_pdf/-char/en |
| work_keys_str_mv | AT tadashiyamazaki chloroquineinhibitsartificialoocyteactivationinducedbyethanolorsr2butnotbysperminmice AT mdwasimbari chloroquineinhibitsartificialoocyteactivationinducedbyethanolorsr2butnotbysperminmice AT satoshikishigami chloroquineinhibitsartificialoocyteactivationinducedbyethanolorsr2butnotbysperminmice |