title: Effect of vitrification on meiotic maturation and expression of genes in immature goat cumulus oocyte complexes.
abstract: The aim of the study was to evaluate meiotic maturation, and expression of genes coding for oocyte secreted factors (GDF9, BMP15, TGFBR1, and BPR2) and apoptosis (BCL2, BAX and P53) after vitrification of immature goat cumulus oocyte complexes (COCs) and in vitro maturation. COCs were vitrified in a solution containing ethylene glycol, dimethyl sulfoxide and sucrose using either a conventional straw (CS), open pulled straw (OPS), cryoloop (CL), hemistraw (HS) or cryotop (CT). Freshly collected COCs (Control), COCs exposed to vitrification and dilution solutions without cryopreservation (EC) and vitrified-warmed COCs were matured in vitro for 27h. The viability of vitrified-warmed COCs 2 h post warming and in vitro maturation was similar for CL, HS and CT. The proportion of oocytes that extruded a 1st polar body and reached TI/MII was significantly higher with CT and HS followed by CL, OPS and CS. Gene expression of GDF9, BMP15, BMPR2, BAX and P53 were comparable to control levels for OPS, CL, HS and CT. The gene expression pattern in CS vitrified COCs was by contrast changed in that GDF9, BMP15, TGFBR1 and BAX were up regulated and BMPR2, BCL2 and P53 down regulated. In conclusion immature goat COCs vitrified using CT and HS showed that viability, maturation rates and expression of genes coding for oocyte secreted factors and apoptosis were similar to non-vitrified controls.
authors.affiliation: Laboratory for Conservation of the Endangered Species (LaCONES), Centre for Cellular and Molecular Biology (CCMB), Hyderabad, India.
authors.forename: Brahmasani Sambasiva
authors.initials: BS
authors.lastname: Rao
authors.forename: Yelisetti Uma
authors.initials: YU
authors.lastname: Mahesh
authors.forename: Katari Venu
authors.initials: KV
authors.lastname: Charan
authors.forename: Komjeti
authors.initials: K
authors.lastname: Suman
authors.forename: Nithya
authors.initials: N
authors.lastname: Sekhar
authors.forename: Sisinthy
authors.initials: S
authors.lastname: Shivaji
mesh_terms: D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology; D054885:Cumulus Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D019855:Ethylene Glycol / Q000494:pharmacology; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006041:Goats; D055412:Growth Differentiation Factors / Q000235:genetics / Q000378:metabolism; D008540:Meiosis / Q000187:drug effects / Q000235:genetics*; D009865:Oocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism; D013395:Sucrose / Q000494:pharmacology; D058989:Vitrification
chemical_list: D051017:Apoptosis Regulatory Proteins; D003451:Cryoprotective Agents; D055412:Growth Differentiation Factors; D018125:Receptors, Transforming Growth Factor beta; D013395:Sucrose; D017346:Protein Serine-Threonine Kinases; D000077293:Receptor, Transforming Growth Factor-beta Type I; D019855:Ethylene Glycol; D004121:Dimethyl Sulfoxide