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Combination of trehalose and low boron in presence of decreased glycerol improves post-thawed ram sperm parameters: A model study in boron research

Andrology · Mart 2022

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YÖKSİS Kayıtları
Combination of trehalose and low boron in presence of decreased glycerol improves post‐thawed ram sperm parameters: A model study in boron research
Andrology · 2021 SCI-Expanded
DOKTOR ÖĞRETİM ÜYESİ MUSTAFA BODU →
Combination of trehalose and low boron in presence of decreased glycerol improves post\u2010thawed ram sperm parameters: A model study in boron research
Andrology · 2022 SCI-Expanded
PROFESÖR NURİ BAŞPINAR →
Combination of trehalose and low boron in presence of decreased glycerol improves post\u2010thawed ram sperm parameters: A model study in boron research
Andrology · 2022 SCI-Expanded
PROFESÖR MUSTAFA NUMAN BUCAK →

Makale Bilgileri

DergiAndrology
Yayın TarihiMart 2022
Cilt / Sayfa10 · 585-594
Erişim🔓 Açık Erişim
Özet Background: Sperm cryopreservation has been widely used in the field of reproductive biotechnology. It applies to certain males of economic and scientific values, including livestock breeds or endangered animal species. The development of a semen extender with a low cryoprotectant concentration and an appropriate amount of trehalose and boron can prevent the deterioration of sperm parameters. Objective: The main goal of this study is to establish a suitable ram extender model, by examining different combinations of high (5%) and low (3%) glycerol concentrations (to reduce its toxic effects on sperm freezing), a fixed amount of trehalose and an increased dose of boron to prevent the deterioration of sperm parameters, and investigate the levels of gene expressions. Materials and methods: The Merino ram ejaculates were collected. The collected ejaculates providing the defined criteria were pooled. The pooled ejaculates were divided into eight aliquots and diluted with the Tris extender including different combinations of glycerol (5% and 3%) and boron (0.25, 0.5, and 1 mm) concentrations and a fixed amount of trehalose, then frozen. After freeze–thawing process, sperm motility, mitochondrial membrane activity, plasma membrane integrity, acrosomal membrane integrity, DNA damage (single cell gel electrophoresis (COMET) and TUNEL assays) as well as NAD(P)H quinone oxyreductase (NQO1), glutamate-cycteine ligase (GCLC), and glutathione S-transferase (GSTP1) for molecular mechanisms of sperm cell response to oxidative stress were assessed for different extender groups following freeze–thawing process: 5% glycerol + 0 mm boron (G5B0.00), 5% glycerol + 0.25 mm boron (G5B0.25), 5% glycerol + 0.5 mm boron (G5B0.50), 5% glycerol + 1 mm boron (G5B1.00), 3% glycerol + 0 mm boron (G3B.00), 3% glycerol + 0.25 mm boron (G3B0.25), 3% glycerol + 0.5 mm boron (G3B0.50), and 3% glycerol + 1 mm boron (G3B1.00). Results: G3B0.25 presented higher percentages of subjective motility, mitochondrial activity, and viability of spermatozoa comparing with G5B0.00 and groups with boron. Supplementation of 0.25 mm boron with and without trehalose (G3B0.25 and G5B0.25) showed higher acrosome integrity, compared with G5B0.00, G5B1.00, G3B0.50, and G3B1.00. For TUNEL analysis, G3B1.00 showed the highest DNA integrity among the experimental groups which was statistically significant only with G5B0.50 (p < 0.05). The mRNA levels of NQO1 were significantly decreased in G5B1.00, G3B0.50, and G3B1.00, when compared to G5B0.00. In comparison with G5B0.00, supplementation of 1 mm boron with and without trehalose had significantly lower expression of GCLC. The level of GSTP1 gene was significantly lower (approximately threefold) in G3B1.00, compared to G5B0.00 (p < 0.05). Discussion and conclusion: It can be assumed that the increase of the boron concentration in the extender may have important adverse effects on sperm parameters and antioxidant gene expression after thawing. The results obtained from this study will help to understand the toxicity limits of boron and eliminate the toxicity of glycerol in studies of gametes and tissue freezing. Therefore, it can be concluded that the use of sufficient boron can decrease cryodamages of cryopreservation of mammalian spermatozoa as well tissue engineering.

Yazarlar (11)

1
Mustafa Numan Bucak
2
Nazan Keskin
3
Mustafa Bodu
ORCID: 0000-0002-8432-7347
4
Bülent Bülbül
5
Mesut Kirbaş
6
Ali Erdem Öztürk
7
Fateme Frootan
8
Pinar Ili
9
Hüseyin Özkan
10
N. Baspinar
11
Şükrü Dursun

Anahtar Kelimeler

boron cryopreservation ram semen trehalose

Kurumlar

Aksaray Üniversitesi
Aksaray Turkey
Bahri Dagdas International Agricultural Research Institute
Konya Turkey
Dokuz Eylül Üniversitesi
Izmir Turkey
Erciyes Üniversitesi
Kayseri Turkey
Mustafa Kemal Üniversitesi
Antakya Turkey
National Institute for Genetic Engineering and Biotechnology Iran
Tehran Iran
Pamukkale Üniversitesi
Denizli Turkey
Pamukkale Üniversitesi Tip Fakültesi
Denizli Turkey
Selçuk Üniversitesi
Selçuklu Turkey

Metrikler

7
Atıf
11
Yazar
4
Anahtar Kelime