Autor(es): Paschoal, Daniela Martins ; Sudano, Mateus José ; Schwarz, Kátia Regina Lancellotti ; Maziero, Rosiára Rosário Dias ; Guastali, Midyan Daroz ; Crocomo, Letícia Ferrari ; Magalhães, Luis Carlos Oña ; Martins, Alício ; Leal, Claudia Lima Verde ; Landim-Alvarenga, Fernanda da Cruz
Data: 2022
Identificador Persistente: http://hdl.handle.net/11449/231388
Origem: Oasisbr
Assunto(s): Apoptosis; Blastocyst; Cryopreservation; Forskolin; Lipid; Vitrification
Made available in DSpace on 2022-04-29T08:45:02Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-01-01
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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 μM. 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 μM: 128.5; P < 0.05), and number of cells per embryo higher (control: 140.1 and F 2.5 μM: 173.5; P < 0.05) than controls for 2.5 μM 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 μM: 101.3, F 5 μM: 115.4, F 10 μM: 95.1; apoptotic of fresh cells, control: 12.1%, F 2.5 μM: 16.7%, F 5 μM: 11.1%, F 10 μM: 14.2%; and apoptotic warmed embryos, control: 22.3%, F 2.5 μM: 37.3%, F 5 μM: 33.2%, F 10 μM: 30.3%; 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.
Departmento de Medicina Veterinária Faculdade de Zootecnia e Engenharia de Alimentos Universidade de São Paulo
Universidade Federal do Pampa Faculdade de Medicina Veterinária
Departmento de Reprodução Animal Faculdade de Medicina Veterinária e Zootecnia Universidade do Estado de São Paulo
Departmento de Clínica Cirurgia e Reprodução Animal Faculdade de Medicina Veterinária Universidade do Estado de São Paulo
FAPESP: 2010/50410-2
FAPESP: 2011/08926-4
FAPESP: 2014/21289-1
