Crotamine, a cell-penetrating peptide, is able to translocate parthenogenetic and in vitro fertilized bovine embryos but does not improve exogenous DNA expression
| Autor: | Luciana Magalhães Melo, Romina J. Bevacqua, Vicente José de Figueirêdo Freitas, Gandhi Rádis-Baptista, Daniel Felipe Salamone, Iana S. Campelo, N. G. Canel |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Otras Biotecnología Agropecuaria animal structures Zygote Parthenogenesis Biotecnología Agropecuaria CATTLE Perivitelline space Cell-Penetrating Peptides Fertilization in Vitro Biology PEPTIDE UPTAKE GENE DELIVERY TRANSGENE EXPRESSION Embryo Culture Techniques Andrology 03 medical and health sciences PREIMPLANTATION EMBRYO Crotalid Venoms Genetics medicine Animals Blastocyst Zona pellucida reproductive and urinary physiology Genetics (clinical) Technological Innovations Obstetrics and Gynecology purl.org/becyt/ford/4.4 [https] Embryo General Medicine Transfection Embryo Mammalian Molecular biology Crotamine 030104 developmental biology medicine.anatomical_structure Reproductive Medicine CIENCIAS AGRÍCOLAS embryonic structures Cattle Female Exogenous DNA purl.org/becyt/ford/4 [https] Developmental Biology |
| Zdroj: | CONICET Digital (CONICET) Consejo Nacional de Investigaciones Científicas y Técnicas instacron:CONICET |
| Popis: | Purpose: Crotamine is capable of penetrating cells and embryos and transfecting cells with exogenous DNA. However, no studies are available regarding its uptake by parthenogenetic (PA) embryos or its use for transfection in in vitro fertilized (IVF) embryos. This study aimed to determine the translocation kinetics of crotamine into PA and IVF bovine embryos and assess its effect over in vitro development of PA embryos. Moreover, crotamine-DNA complexes were used to test the transfection ability of crotamine in bovine IVF zygotes. Methods: PA and IVF embryos were exposed to labeled crotamine for four interval times. Embryo toxicity was assayed over PA embryos after 24 h of exposure to crotamine. Additionally, IVF embryos were exposed to or injected with a complex formed by crotamine and pCX-EGFP plasmid. Results: Confocal images revealed that crotamine was uptaken by PA and IVF embryos as soon as 1 h after exposure. Crotamine exposure did not affect two to eight cells and blastocyst rates or blastocyst cell number (p > 0.05) of PA embryos. Regarding transfection, exposure or injection into the perivitelline space with crotamine-DNA complex did not result in transgene-expressing embryos. Nevertheless, intracytoplasmic injection of plasmid alone showed higher expression rates than did injection with crotamine-DNA complex at days 4 and 7 (p < 0.05). Conclusions: Crotamine is able to translocate through zona pellucida (ZP) of PA and IVF embryos within 1 h of exposure without impairing in vitro development. However, the use of crotamine does not improve exogenous DNA expression in cattle embryos, probably due to the tight complexation of DNA with crotamine. Fil: Campelo, Iana S.. State University of Ceará; Brasil Fil: Canel, Natalia Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina Fil: Bevacqua, Romina Jimena. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina Fil: Melo, Luciana M.. State University of Ceará; Brasil Fil: Rádis Baptista, Gandhi. State University of Ceará; Brasil Fil: Freitas, Vicente J. F.. State University of Ceará; Brasil Fil: Salamone, Daniel Felipe. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Agronomía. Pabellón de Zootecnica. Laboratorio de Biotecnología Animal; Argentina |
| Databáze: | OpenAIRE |
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