Cell apoptosis and lipid content of in vitro-produced, vitrified bovine embryos treated with forskolin

Author(s)

Paschoal, Daniela Martins ; Sudano, Mateus Jose ; Lancellotti Schwarz, Katia Regina ; Dias Maziero, Rosiara Rosario ; Guastali, Midyan Daroz ; Crocomo, Leticia Ferrari ; Ona Magalhaes, Luis Carlos ; Martins, Jr., Alicio ; Verde Leal, Claudia Lima ; Landim-Alvarenga, Fernanda da Cruz

• Document type: Journal article
• Source: Theriogenology; v. 87, p. 108-114, JAN 1 2017.
• Web of Science Citations: 14

Abstract

The presence of fetal calf serum in culture medium influences embryo quality, causing a reduction in postcryopreservation survival. Forskolin has been used to induce lipolysis and increase cryotolerance, functioning as an activator of adenylate cyclase and elevating CAMP levels. In the present experiment, bovine zygotes were cultured in synthetic oviduct fluid with amino acid plus 2.5% fetal calf serum for 6 days, when forskolin was added in three concentrations: 2.5, 5, and 10 mu M. Treatment with forskolin lasted for 24 hours. Blastocyst formation rate, quantification of lipid granules, total cell numbers, and apoptosis rate were evaluated. In a second assessment, embryos were vitrified, and warming, re-expansion rate, total cell numbers, and apoptosis rate were also evaluated. There was no difference due to forskolin in blastocyst formation or re-expansion rates after vitrification. However, lipid measurements were lower (control: 136.8 and F 2.5 mu M: 128.5; P < 0.05), and number of cells per embryo higher (control: 140.1 and F 2.5 mu M: 173.5; P < 0.05) than controls for 2.5 M forskolin but not for higher forskolin concentrations. The number of intact cells per embryo was higher, and the rate of apoptosis was lower in fresh than in vitrified embryos (number of cells of warmed embryos, control: 104.1, F 2.5 mu M: 101.3, F 5 mu M: 115.4, F 10 mu M: 95.1; apoptotic of fresh cells, control: 12.1%, F 2.5 mu M: 16.7%, F 5 mu M: 11.1%, F 10 mu M: 14.2%; and apoptotic warmed embryos, control: 22.3%, F 2.5 mu M: 373%, F 5 mu M: 33.2%, F 10 mu M: 303%; P < 0.05). It was concluded that forskolin is an effective lipolytic agent even at low concentrations, leading to formation of blastocysts with a comparatively larger number of cells. (C) 2016 Elsevier Inc. All rights reserved. (AU)

• FAPESP's process: 14/21289-1 - Temporary inhibition of meiosis and chemical lipolysis in oocyte: innovations in bovine embryos in vitro production
• Grantee: Daniela Martins Paschoal
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 11/08926-4 - Effects of forskolin, butyrolactone I and roscovitine in maturation of bovine oocytes and its implications in resistence to cryopreservation
• Grantee: Fernanda da Cruz Landim
• Support Opportunities: Regular Research Grants