Effect of Protein Sources on Glutathione Peroxidase Enzyme Activity of In Vitro Maturated Bovine Oocytes

Nisa Nur Yılmaz

Makale detayları

Effect of Protein Sources on Glutathione Peroxidase Enzyme Activity of In Vitro Maturated Bovine Oocytes

Yazarlar:

Nisa Nur Yılmaz

Yayıncı:

Karyay Karadeniz Yayımcılık

Ana Başlıklar:

Hayvan Gelişim ve Üreme Biyolojisi

Dergi:

Black Sea Journal of Engineering and Science Vol.7 Issue 3, pp.575-579

Belge Tipi:

Makale

Yayın Tarihi:

5/2024

Belge Dili:

Türkçe

Document ID (Org No.):

2001315144

pISSN:

2619-8991

Anahtar kelimeler:

Özet:

 This study was conducted to determine the effect of protein sources added to the culture media during in vitro maturation IVM on the glutathione peroxidase GPx enzyme activity of bovine oocytes. Oocytes obtained from bovine ovaries were IVM cultured in bicarbonate buffered tissue culture media TCM-199 containing 10 fetal calf serum FBS, 4 mgmL bovine serum albumin SSA, 1 mgmL polyvinyl alcohol PVA and without protein source for 22 hours in a humidified environment containing 38.5 C and 5 CO2. Following the maturation period, cumulus-oocyte complexes COCs obtained were subjected to a procedure to separate the cumulus cells from the oocytes. The oocytes separated from the cumulus cells were washed with phosphate-buffered saline PBS containing polyvinyl alcohol and devoid of Ca2 and Mg2. Subsequently, the oocytes were divided into predetermined sample sizes and stored in microtubes for further processing. After IVM culture, using a commercial kit, glutathione peroxidase GPx enzyme activity in oocytes was determined spectrophotometrically at a wavelength of 340 nm. In the study, the levels of GPx enzyme activity in the cell extracts isolated from the oocytes in each experimental group did not change over time, and the GPx enzyme activity in the oocytes with PVA added to the culture medium was lower than the oocytes in the other experimental groups P0.05. As a result of this study, the effects of different protein sources on GPx enzyme levels, one of the indicators of antioxidant activity in bovine oocytes, were determined, and FBS and BSA, as protein sources, could be more effective on bovine oocyte development.

 This study was conducted to determine the effect of protein sources added to the culture media during in vitro maturation IVM on the glutathione peroxidase GPx enzyme activity of bovine oocytes. Oocytes obtained from bovine ovaries were IVM cultured in bicarbonate buffered tissue culture media TCM-199 containing 10 fetal calf serum FBS, 4 mgmL bovine serum albumin SSA, 1 mgmL polyvinyl alcohol PVA and without protein source for 22 hours in a humidified environment containing 38.5 C and 5 CO2. Following the maturation period, cumulus-oocyte complexes COCs obtained were subjected to a procedure to separate the cumulus cells from the oocytes. The oocytes separated from the cumulus cells were washed with phosphate-buffered saline PBS containing polyvinyl alcohol and devoid of Ca2 and Mg2. Subsequently, the oocytes were divided into predetermined sample sizes and stored in microtubes for further processing. After IVM culture, using a commercial kit, glutathione peroxidase GPx enzyme activity in oocytes was determined spectrophotometrically at a wavelength of 340 nm. In the study, the levels of GPx enzyme activity in the cell extracts isolated from the oocytes in each experimental group did not change over time, and the GPx enzyme activity in the oocytes with PVA added to the culture medium was lower than the oocytes in the other experimental groups P0.05. As a result of this study, the effects of different protein sources on GPx enzyme levels, one of the indicators of antioxidant activity in bovine oocytes, were determined, and FBS and BSA, as protein sources, could be more effective on bovine oocyte development.

 This study was conducted to determine the effect of protein sources added to the culture media during in vitro maturation IVM on the glutathione peroxidase GPx enzyme activity of bovine oocytes. Oocytes obtained from bovine ovaries were IVM cultured in bicarbonate buffered tissue culture media TCM-199 containing 10 fetal calf serum FBS, 4 mgmL bovine serum albumin SSA, 1 mgmL polyvinyl alcohol PVA and without protein source for 22 hours in a humidified environment containing 38.5 C and 5 CO2. Following the maturation period, cumulus-oocyte complexes COCs obtained were subjected to a procedure to separate the cumulus cells from the oocytes. The oocytes separated from the cumulus cells were washed with phosphate-buffered saline PBS containing polyvinyl alcohol and devoid of Ca2 and Mg2. Subsequently, the oocytes were divided into predetermined sample sizes and stored in microtubes for further processing. After IVM culture, using a commercial kit, glutathione peroxidase GPx enzyme activity in oocytes was determined spectrophotometrically at a wavelength of 340 nm. In the study, the levels of GPx enzyme activity in the cell extracts isolated from the oocytes in each experimental group did not change over time, and the GPx enzyme activity in the oocytes with PVA added to the culture medium was lower than the oocytes in the other experimental groups P0.05. As a result of this study, the effects of different protein sources on GPx enzyme levels, one of the indicators of antioxidant activity in bovine oocytes, were determined, and FBS and BSA, as protein sources, could be more effective on bovine oocyte development.

 Bu çalışma, sığır oositlerinin in vitro olgunlaştırma IVOsında kültür medyumuna ilave edilen protein kaynaklarının glutatyon peroksidaz GPx enzim aktivitesi üzerine etkisinin belirlenmesi amacıyla yapılmıştır. Sığır ovaryumlarından elde edilen oositler 10 fötal buzağı serum FBS, 4 mgmL sığır serum albümin SSA, 1 mgmL polivinil alkol PVA içeren ve protein kaynağı içermeyen bikarbonat tamponlu doku kültür medyumlarında TCM199 38,5 C sıcaklıkta ve 5 CO2 içeren nemlendirilmiş ortamda 22 saat IVOya alınmıştır. Olgunlaşma süresi sonunda elde edilen kumulus-oosit kompleksleri KOKler, kumulus hücrelerinden ayırmak için işleme tabi tutulmuştur. Kumulus hücrelerinden ayrılan oositler, polivinil alkol içeren ve Ca2 ve Mg2 içermeyen fosfat tamponlu tuz çözeltisi PBS ile yıkanmıştır. Daha sonra, oositler belirlenmiş örnek boyutlarına bölünerek mikrotüplerde saklanmıştır. IVO kültürü sonrasında oositlerdeki glutatyon peroksidaz GPx enzim aktivitesi için ticari kit kullanarak 340 nm dalga boyunda spektrofotometrik olarak belirlenmiştir. Çalışmada, her bir deneme grubundaki oositlerden izole edilen hücre ekstraktlarındaki GPx enzim aktivitesinin seviyelerinin zamanla değişmediği, kültür medyumuna PVA eklenmiş oositlerinde GPx enzim aktivitesinin diğer deneme gruplarındaki oositlerden daha düşük olduğu tespit edilmiştir P0,05. Bu çalışma sonucunda farklı protein kaynaklarının antioksidan aktivitenin göstergelerinden biri olan GPx enzim seviyelerinin sığır oositlerinde üzerine etkileri belirlenmiş ve FBS ve BSAnın protein kaynağı olarak sığır oosit gelişimi üzerine daha etkili olabileceği tespit edilmiştir.

Makale detayları

Effect of Protein Sources on Glutathione Peroxidase Enzyme Activity of In Vitro Maturated Bovine Oocytes

Yazarlar:

Nisa Nur Yılmaz

Yayıncı:

Karyay Karadeniz Yayımcılık

Ana Başlıklar:

Hayvan Gelişim ve Üreme Biyolojisi

Dergi:

Black Sea Journal of Engineering and Science Vol.7 Issue 3, pp.575-579

Belge Tipi:

Makale

Yayın Tarihi:

5/2024

Belge Dili:

Türkçe

Document ID (Org No.):

2001315144

pISSN:

2619-8991

Oocyte

Bovine

Antioxidant activity

Protein

MaturationOosit

Sığır

Antioksidan aktivitesi

Protein

Olgunlaşma

Sayı

Makale detayları

Yazarlar: Nisa Nur Yılmaz

Yayıncı: Karyay Karadeniz Yayımcılık

Ana Başlıklar: Hayvan Gelişim ve Üreme Biyolojisi

Dergi: Black Sea Journal of Engineering and Science Vol.7 Issue 3, pp.575-579

Belge Tipi: Makale

Yayın Tarihi: 5/2024

Belge Dili: Türkçe

Document ID (Org No.): 2001315144

pISSN: 2619-8991

Sayı

Yazarlar:

Nisa Nur Yılmaz

Yayıncı:

Karyay Karadeniz Yayımcılık

Ana Başlıklar:

Hayvan Gelişim ve Üreme Biyolojisi

Dergi:

Black Sea Journal of Engineering and Science Vol.7 Issue 3, pp.575-579

Belge Tipi:

Makale

Yayın Tarihi:

5/2024

Belge Dili:

Türkçe

Document ID (Org No.):

2001315144

pISSN:

2619-8991