Expression profile of key genes involved in DNA repair mechanisms in bovine cumulus cells cultured with bovine serum albumin or fetal calf serum

被引:1
|
作者
Goetten, Andre Lucio Fontana [1 ]
Koch, Julia [1 ]
Rocha, Cecilia Constantino [2 ]
Mezzalira, Alceu [3 ]
Price, Christopher Allan [4 ]
Portela, Valerio Marques [5 ]
Barreta, Marcos Henrique [1 ]
机构
[1] Univ Fed Santa Catarina UFSC, Ctr Ciencias Rurais, Campus Curitibanos, Rodovia Ulysses Gaboardi, Km 0, BR-89520000 Curitibanos, SC, Brazil
[2] Univ Florida, Dept Anim Sci, Gainesville, FL USA
[3] Univ Estado Santa Catarina UDESC, Ctr Ciencias Agrovet CAV, Lab Reprod Anim Assis Roberto Bem, Lages, SC, Brazil
[4] Univ Montreal, Fac Med Vet, Ctr Rech Reprod Anim, St Hyacinthe, PQ, Canada
[5] Univ Fed Santa Maria UFSM, Lab Biotecnol & Reprod Anim BIOREP, Santa Maria, RS, Brazil
关键词
In vitro maturation; Cumulus cells; DNA repair; Apoptosis; IN-VITRO MATURATION; MORPHOGENETIC PROTEIN 15; OOCYTE COMPLEXES; HOMOLOGOUS RECOMBINATION; DAMAGE RESPONSE; EMBRYO QUALITY; ROLES; EXPANSION; 53BP1; END;
D O I
10.1016/j.repbio.2022.100709
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
Cumulus cells from cumulus-oocyte complexes (COC) matured in vitro in serum-free medium show high inci-dence of apoptosis and DNA double-strand breaks (DSB). This study aimed to characterize the transcript expression profile of selected genes involved in DNA repair mechanisms in bovine cumulus cells cultured with bovine serum albumin (BSA) or fetal calf serum (FCS). Briefly, bovine cumulus-oocyte complexes were in vitro matured with either, 0.4% BSA or 10% FCS for 3, 6, 12 or 24 h. The total RNA of cumulus cells was used for real-time PCR analysis. Transcript abundance of XRCC6, XRCC5, DNAPK, GAAD45B, TP53BP1, RAD50, RAD52, ATM and BRCA2 target genes changed as the IVM proceeded (P < 0.05). However, an interaction between protein source (FCS or BSA) and time was not detected (P >= 0.05). Cumulus cells from COCs matured with BSA presented higher mRNA expression of two genes compared to FCS group: TP53BP1 at 6 h and BRCA1 at 3, 6, 12 and 24 h (P < 0.05). In summary, our results showed for the first time the expression profile of the key genes involved in DSB repair mechanisms in cumulus cells obtained from bovine COCs matured with FCS or BSA. The higher mRNA expression of BRCA1 and TP53BP1 and lower mRNA expression of TNFAIP6 suggests an increase in apoptosis rate and DNA damage in cumulus cells cultured in BSA-supplemented medium and may explain, at least to some extent, the reduced developmental potential of bovine oocytes matured in serum-free medium.
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页数:9
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