Hydrogen peroxide concentration and DNA fragmentation of buffalo oocytes matured in sericin-supplemented maturation medium

Abstract

This study was conducted to investigate the effects of sericin on meiotic maturation, hydrogen peroxide (H 2 O 2) concentration and deoxyribonucleic acid (DNA) fragmentation in buffalo oocytes. Oocytes were matured in vitro in tissue culture medium (TCM-199) with in vitro maturation (IVM) groups under several conditions, namely without bovine serum albumin (-BSA), (+BSA), and 0.025%, 0.05%, 0.1% and 0.25% (w/v) sericin. The results showed that supplementation of the maturation medium with 0.05% sericin significantly increased the rate of oocytes that reach metaphase II compared with other groups, except for the 0.025% sericin-treated group. Intracellular H 2 O 2 concentrations in oocytes of all groups were measured using 2',7'-dichlorodihydrofluorescein diacetate (DCHFDA). The concentration of H 2 O 2 in matured oocytes treated with 0.05% sericin was lower than in other groups. DNA fragmentation and the nuclear status of oocytes were examined using the terminal deoxyribonucleotidyl transferase-mediated dUTP digoxigenin nick end-labelling (TUNEL) method. The total proportion of TUNEL-positive oocytes at the MII stage was lower in the 0.05% sericin group. The results indicate that addition of 0.05% sericin to the maturation medium may improve nuclear maturation and may inhibit DNA fragmentation in oocytes by decreasing H 2 O 2 concentrations.

Habitat Preferences of Wild Mammalian Species around River Chenab in Sialkot, Gujrat and Gujranwala Districts, Punjab, Pakistan

“…Embryos that have been re-cultured for 48 h, namely control and embryos that have been grouped based on the stages of division, then concentration measurements are carried out H 2 O 2 concentration was measured using 2',7'-dichlorodihydro fluorescein diacetate (DCHFDA) (Sigma) as described by Gustina et al (2019) with minor modifications. Embryos in each group were incubated for 15 min with a culture medium containing 10 μM of DCHFDA, were washed in a fresh medium before placed on a glass slide and were covered by a cover glass.…”

“…An increase in H 2 O 2 can be detrimental not only to the success rate of inductive assisted reproduction but also result in epigenetic and genetic changes in the embryo, which have an effect on transgeneration (Hardy et al, 2021). Stress tolerance after H 2 O 2 exposure is not mediated by the use of antioxidants such as glutathione (Vandaele et al, 2010) melantonin (Tamura et al, 2012), sericin (Gustina et al, 2019) able to block ROS (Cavallari et al, 2019). The weakness of this study is that the concentration of H 2 O 2 in oocytes after ripening and the concentration of H 2 O 2 in Bali cattle sperm have not been measured.…”

Study of the Antioxidative Effects of Bombyx mori Silk Sericin in Cultures of Murine Retinal Photoreceptor Cells

“…BMSS has been reported as an antioxidant in preparations isolated directly from cocoons, generally followed by additional purification, and then assessing its antioxidant capacity either by a variety of chemical assays [ 31 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 65 ] or in cell cultures [ 61 , 63 , 65 , 66 , 67 , 68 , 69 , 70 ]. Antioxidant activity has also been demonstrated in the non-peptidoid compounds associated with sericin that were isolated by various separation techniques [ 60 , 65 , 71 , 72 , 73 , 74 ].…”

Improved Maturation Rate of Bovine Oocytes Following Sericin Supplementation in Collection and Maturation Media

“…Chlapanidas et al (2013) report that sericin is often supplemented in the culture medium. Sericin as an antioxidant prevents March 2022 25 a stress oxidative in the oocytes by decreasing the production of hydrogen peroxide (H 2 O 2 ) (Isobe et al, 2014;Gustina et al, 2019) and inhibiting lipid peroxidation (Kato et al, 1998). The hydroxyl groups are responsible for the antioxidant action by chelating trace elements such as copper and iron (Kato et al, 1998).…”

“…The hydroxyl groups are responsible for the antioxidant action by chelating trace elements such as copper and iron (Kato et al, 1998). In IVEP, supplementation of sericin in the maturation medium increases the nuclear maturation of oocytes of buffalo (Gustina et al, 2017;Gustina et al, 2019) and sheep (Yasmin et al, 2015). The process of oocyte collection consists of the COCs aspiration of all ovaries, precipitation of aspirated fluid, and grading of the oocytes under a microscope.…”