Effect of zinc chloride and sodium selenite supplementation on in vitro maturation, oxidative biomarkers, and gene expression in buffalo (Bubalus bubalis) oocytes

Abstract

Summary This study examined the effects of zinc chloride (ZnCl2) and sodium selenite (Na2SeO3) supplementation in maturation medium on in vitro maturation (IVM) rate, oxidative biomarkers and gene expression in buffalo oocytes. Ovaries from a slaughterhouse were aspirated and good quality cumulus–oocyte complexes (COCs) with at least four layers of compact cumulus cells and evenly granulated dark ooplasm were selected. COCs were randomly allocated during IVM (22 h) to one of four treatment groups: (1) control maturation medium (basic medium), or basic medium supplemented with (2) ZnCl2 (1.5 µg/ml), (3) Na2SeO3 (5 µg/l), or (4) ZnCl2 + Na2SeO3 (1.5 µg/ml + 5 µg/l, respectively). Oocytes were denuded after 22 h of IVM in the first four replicates. Specimens were fixed and stained to evaluate the stage of nuclear maturation. The spent medium was collected for biochemical assays of total antioxidant capacity (TAC), malondialdehyde (MDA) and hydrogen peroxide concentrations. A second four replicates were used for COCs for RNA extraction. The expression levels of antioxidant (SOD1, GPX4, CAT and PRDX1), antiapoptotic (BCL2 and BCL-XL) and proapoptotic (BAX and BID) genes were measured. Supplementation with ZnCl2 and Na2SeO3 during IVM increased the ratio of oocytes reaching metaphase II at 22 h, increased TAC and decreased MDA and H2O2 concentrations in the maturation medium (P < 0.05). Moreover, beneficial effects were associated with complementary changes in expression patterns of antioxidative, antiapoptotic and proapoptotic genes, suggesting lower oxidative stress and apoptosis. Supplementation medium with zinc chloride and sodium selenite improves the maturation rate, reduces oxidative stress and increases expression levels of antioxidative and antiapoptotic genes.

Effect of in Vitro Maturation Medium Supplementation with Chitosan Nanoparticles on The River Buffalo Cumulus-Oocyte Complexes

“…These included SOD1, BCL2, BAX and B actin. The SOD1 had accession number of NM_001290973.1, product length of153bp, with primer sequences of F-GAGAGGCATGTTGGAGACCT and R -TCTGCCCAAGTCATCTGGTT (Khalil et al 2021). For BCL2, the accession number was XM_025273634.1, product length was110bp, with primer sequences of F-ATGACTTCTCTCGGCGCTAC and R-TGAAGAGCTC CTCCACCAC (Khalil et al 2021).…”

“…The SOD1 had accession number of NM_001290973.1, product length of153bp, with primer sequences of F-GAGAGGCATGTTGGAGACCT and R -TCTGCCCAAGTCATCTGGTT (Khalil et al 2021). For BCL2, the accession number was XM_025273634.1, product length was110bp, with primer sequences of F-ATGACTTCTCTCGGCGCTAC and R-TGAAGAGCTC CTCCACCAC (Khalil et al 2021). BAX had accession number of XM_006050927.2, product length 122bp and primer sequences of F-CTTTTGCTTCAGGGTTTCA and R-CGCTTCAGACACTCGCTCA (Khalil et al 2021).…”

“…For BCL2, the accession number was XM_025273634.1, product length was110bp, with primer sequences of F-ATGACTTCTCTCGGCGCTAC and R-TGAAGAGCTC CTCCACCAC (Khalil et al 2021). BAX had accession number of XM_006050927.2, product length 122bp and primer sequences of F-CTTTTGCTTCAGGGTTTCA and R-CGCTTCAGACACTCGCTCA (Khalil et al 2021). For B actin, accession number was XM_006044278.1, product length was 150bp, the primer sequences were F-GCCCTGGCACCCAGCACAAT and R-GGAGGGGCC GGACTCATCGT (Zhao et al 2019).…”