title: Effect of crotamine, a cell-penetrating peptide, on blastocyst production and gene expression of in vitro fertilized bovine embryos.
abstract: The present study investigated the effects of crotamine, a cell-penetrating peptide from rattlesnake venom, at different exposure times and concentrations, on both developmental competence and gene expression (ATP1A1, AQP3, GLUT1 and GLUT3) of in vitro fertilized (IVF) bovine embryos. In Experiment 1, presumptive zygotes were exposed to 0.1 μM crotamine for 6, 12 or 24 h and control groups (vehicle and IVF) were included. In Experiment 2, presumptive zygotes were exposed to 0 (vehicle), 0.1, 1 and 10 μM crotamine for 24 h. Additionally, to visualize crotamine uptake, embryos were exposed to rhodamine B-labelled crotamine and subjected to confocal microscopy. In Experiment 1, no difference (P > 0.05) was observed among different exposure times and control groups for cleavage and blastocyst rates and total cells number per blastocyst. Within each exposure time, mRNA levels were similar (P > 0.05) in embryos cultured with or without crotamine. In Experiment 2, concentrations as high as 10 μM crotamine did not affect (P > 0.05) the blastocyst rate. Crotamine at 0.1 and 10 μM did not alter mRNA levels when compared with the control (P > 0.05). Remarkably, only 1 μM crotamine decreased both ATP1A1 and AQP3 expression levels relative to the control group (P < 0.05). Also, it was possible to visualize the intracellular localization of crotamine. These results indicate that crotamine can translocate intact IVF bovine embryos and its application in the culture medium is possible at concentrations from 0.1-10 μM for 6-24 h.
authors.affiliation: Laboratory of Physiology and Control of Reproduction,Faculty of Veterinary,State University of Ceará,Fortaleza-CE,Brazil.
authors.forename: Iana S
authors.initials: IS
authors.lastname: Campelo
authors.affiliation: Laboratory of Physiology and Control of Reproduction,Faculty of Veterinary,State University of Ceará,Fortaleza-CE,Brazil.
authors.forename: Alexsandra F
authors.initials: AF
authors.lastname: Pereira
authors.affiliation: Laboratory of Physiology and Control of Reproduction,Faculty of Veterinary,State University of Ceará,Fortaleza-CE,Brazil.
authors.forename: Agostinho S
authors.initials: AS
authors.lastname: Alcântara-Neto
authors.affiliation: Laboratory of Animal Biotechnology,University of Buenos Aires,Buenos Aires,Argentina.
authors.forename: Natalia G
authors.initials: NG
authors.lastname: Canel
authors.affiliation: Laboratory of Physiology and Control of Reproduction,Faculty of Veterinary,State University of Ceará,Fortaleza-CE,Brazil.
authors.forename: Joanna M G
authors.initials: JM
authors.lastname: Souza-Fabjan
authors.affiliation: Laboratory of Physiology and Control of Reproduction,Faculty of Veterinary,State University of Ceará,Fortaleza-CE,Brazil.
authors.forename: Dárcio I A
authors.initials: DI
authors.lastname: Teixeira
authors.affiliation: Embrapa Dairy Cattle,Juiz de Fora-MG,Brazil.
authors.forename: Luiz S A
authors.initials: LS
authors.lastname: Camargo
authors.affiliation: Laboratory of Physiology and Control of Reproduction,Faculty of Veterinary,State University of Ceará,Fortaleza-CE,Brazil.
authors.forename: Luciana M
authors.initials: LM
authors.lastname: Melo
authors.affiliation: Laboratory of Biochemistry and Biotechnology,Institute of Marine Science,Federal University of Ceará,Fortaleza-CE,Brazil.
authors.forename: Gandhi
authors.initials: G
authors.lastname: Rádis-Baptista
authors.affiliation: Laboratory of Animal Biotechnology,University of Buenos Aires,Buenos Aires,Argentina.
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Salamone
authors.affiliation: Universidade Estadual do Ceará/Faculdade de Veterinária;Av. Dr. Silas Munguba,1700-Fortaleza-CE,60714-903Brazil.
authors.forename: Vicente J F
authors.initials: VJ
authors.lastname: Freitas
mesh_terms: D000818:Animals; D051400:Aquaporin 3 / Q000235:genetics; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D002417:Cattle; D003435:Crotalid Venoms / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000494:pharmacology*; D005260:Female; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D051272:Glucose Transporter Type 1 / Q000235:genetics; D051274:Glucose Transporter Type 3 / Q000235:genetics; D008297:Male; D000254:Sodium-Potassium-Exchanging ATPase / Q000235:genetics
chemical_list: D003435:Crotalid Venoms; D051272:Glucose Transporter Type 1; D051274:Glucose Transporter Type 3; D051400:Aquaporin 3; C010563:crotamine; D000254:Sodium-Potassium-Exchanging ATPase
keywords: Cattle; Developmental competence; Embryotoxicity; Preimplantation embryo; qPCR