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ZnSO 4 induces zinc overload in the oocytes of cultured neonatal mouse ovaries in the presence <t>of</t> <t>ISCK03.</t> Three-days-postpartum mouse ovaries were cultured without (control) or with ZnSO 4 , ISCK03, and/or <t>KITL</t> for 12 h ( C – H ) or 24 h ( G , H ). ( A ) The mRNA levels of zinc homeostasis-related genes expressed in the ovaries were normalized using Zip3 as the reference standard, n = 3, each from six ovaries. ( B ) The mRNA levels of zinc homeostasis-related genes in oocytes and granulosa cells, n = 3, each from three ovaries. OO, oocyte; GC, granulosa cell. ( C ) The mRNA levels of zinc homeostasis-related genes in different treated ovaries, n = 3, each from six ovaries. ( D ) The mRNA levels of Zip6 in different treated ovaries, n = 3, each from six ovaries. ( E ) The protein levels of ZIP6 in different treated ovaries, n = 3, each from six ovaries. ( F ) Localization of ZIP6 in the ovaries (brown). ( G , H ) The relative fluorescence staining (green, ( G )) and fluorescence intensity ( H ) of zinc in the oocytes under different treatments, n = 30–60, each from one oocyte. The representative images are presented. Scale bars, 50 µm. Bars indicate the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.
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ZnSO 4 induces zinc overload in the oocytes of cultured neonatal mouse ovaries in the presence <t>of</t> <t>ISCK03.</t> Three-days-postpartum mouse ovaries were cultured without (control) or with ZnSO 4 , ISCK03, and/or <t>KITL</t> for 12 h ( C – H ) or 24 h ( G , H ). ( A ) The mRNA levels of zinc homeostasis-related genes expressed in the ovaries were normalized using Zip3 as the reference standard, n = 3, each from six ovaries. ( B ) The mRNA levels of zinc homeostasis-related genes in oocytes and granulosa cells, n = 3, each from three ovaries. OO, oocyte; GC, granulosa cell. ( C ) The mRNA levels of zinc homeostasis-related genes in different treated ovaries, n = 3, each from six ovaries. ( D ) The mRNA levels of Zip6 in different treated ovaries, n = 3, each from six ovaries. ( E ) The protein levels of ZIP6 in different treated ovaries, n = 3, each from six ovaries. ( F ) Localization of ZIP6 in the ovaries (brown). ( G , H ) The relative fluorescence staining (green, ( G )) and fluorescence intensity ( H ) of zinc in the oocytes under different treatments, n = 30–60, each from one oocyte. The representative images are presented. Scale bars, 50 µm. Bars indicate the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.
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ZnSO 4 induces zinc overload in the oocytes of cultured neonatal mouse ovaries in the presence <t>of</t> <t>ISCK03.</t> Three-days-postpartum mouse ovaries were cultured without (control) or with ZnSO 4 , ISCK03, and/or <t>KITL</t> for 12 h ( C – H ) or 24 h ( G , H ). ( A ) The mRNA levels of zinc homeostasis-related genes expressed in the ovaries were normalized using Zip3 as the reference standard, n = 3, each from six ovaries. ( B ) The mRNA levels of zinc homeostasis-related genes in oocytes and granulosa cells, n = 3, each from three ovaries. OO, oocyte; GC, granulosa cell. ( C ) The mRNA levels of zinc homeostasis-related genes in different treated ovaries, n = 3, each from six ovaries. ( D ) The mRNA levels of Zip6 in different treated ovaries, n = 3, each from six ovaries. ( E ) The protein levels of ZIP6 in different treated ovaries, n = 3, each from six ovaries. ( F ) Localization of ZIP6 in the ovaries (brown). ( G , H ) The relative fluorescence staining (green, ( G )) and fluorescence intensity ( H ) of zinc in the oocytes under different treatments, n = 30–60, each from one oocyte. The representative images are presented. Scale bars, 50 µm. Bars indicate the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.
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ZnSO 4 induces zinc overload in the oocytes of cultured neonatal mouse ovaries in the presence of ISCK03. Three-days-postpartum mouse ovaries were cultured without (control) or with ZnSO 4 , ISCK03, and/or KITL for 12 h ( C – H ) or 24 h ( G , H ). ( A ) The mRNA levels of zinc homeostasis-related genes expressed in the ovaries were normalized using Zip3 as the reference standard, n = 3, each from six ovaries. ( B ) The mRNA levels of zinc homeostasis-related genes in oocytes and granulosa cells, n = 3, each from three ovaries. OO, oocyte; GC, granulosa cell. ( C ) The mRNA levels of zinc homeostasis-related genes in different treated ovaries, n = 3, each from six ovaries. ( D ) The mRNA levels of Zip6 in different treated ovaries, n = 3, each from six ovaries. ( E ) The protein levels of ZIP6 in different treated ovaries, n = 3, each from six ovaries. ( F ) Localization of ZIP6 in the ovaries (brown). ( G , H ) The relative fluorescence staining (green, ( G )) and fluorescence intensity ( H ) of zinc in the oocytes under different treatments, n = 30–60, each from one oocyte. The representative images are presented. Scale bars, 50 µm. Bars indicate the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Journal: Antioxidants

Article Title: Granulosa Cell-Secreted KITL Is Involved in Maintaining Zinc Homeostasis in the Oocytes of Neonatal Mouse Ovaries

doi: 10.3390/antiox14111345

Figure Lengend Snippet: ZnSO 4 induces zinc overload in the oocytes of cultured neonatal mouse ovaries in the presence of ISCK03. Three-days-postpartum mouse ovaries were cultured without (control) or with ZnSO 4 , ISCK03, and/or KITL for 12 h ( C – H ) or 24 h ( G , H ). ( A ) The mRNA levels of zinc homeostasis-related genes expressed in the ovaries were normalized using Zip3 as the reference standard, n = 3, each from six ovaries. ( B ) The mRNA levels of zinc homeostasis-related genes in oocytes and granulosa cells, n = 3, each from three ovaries. OO, oocyte; GC, granulosa cell. ( C ) The mRNA levels of zinc homeostasis-related genes in different treated ovaries, n = 3, each from six ovaries. ( D ) The mRNA levels of Zip6 in different treated ovaries, n = 3, each from six ovaries. ( E ) The protein levels of ZIP6 in different treated ovaries, n = 3, each from six ovaries. ( F ) Localization of ZIP6 in the ovaries (brown). ( G , H ) The relative fluorescence staining (green, ( G )) and fluorescence intensity ( H ) of zinc in the oocytes under different treatments, n = 30–60, each from one oocyte. The representative images are presented. Scale bars, 50 µm. Bars indicate the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: In treatment groups, the ovaries were cultured in medium supplemented with ZnSO 4 (0–40 μM), ISCK03 (0–5 μM; HY-101038, MedChemExpress, Monmouth Junction, NJ, USA), and/or KITL (8.06 nM; HY-P7064, MedChemExpress).

Techniques: Cell Culture, Control, Fluorescence, Staining