RESUMEN
Our objective was to investigate the effects of in vitro culture (IVC) medium supplemented with amphiregulin (AREG) on the preimplantation embryonic development of porcine (Genus: Sus domestica, Species: Landrace) embryos derived from in vitro fertilization (IVF) and parthenogenetic activation (PA). In vitro fertilization and PA embryos at the 1-cell stage were cultured in IVC medium supplemented with 0, 0.5, 5, or 50 ng/mL AREG for 7 d. There were significantly greater total numbers of cells in blastocysts of IVF and PA embryos cultured with 50 ng/mL AREG compared with that of controls. In vitro fertilization and PA embryos were then cultured in NCSU-23 medium supplemented with 50 ng/mL AREG on Days 1 through 7, Days 1 through 3 (early stage), or Days 4 through 7 (late stage), or without AREG. There were significantly greater numbers of trophoblast cells in the late-stage and full-term groups of IVF and PA embryos than in the early-stage and control groups. The presence of AREG protein in IVF-derived blastocysts was detected using a polyclonal AREG antibody and indirect immunofluorescence. Amphiregulin protein was localized in both the cytoplasm and nucleus. Using real-time polymerase chain reaction, we detected the expression of AREG mRNA in all developmental stages of IVF and PA embryos; however, the expression level varied according to stage. Thus, the incubation of porcine IVF and PA embryos in AREG-supplemented culture medium mainly at the late preimplantation stage increases the numbers of trophoblast cells.
Asunto(s)
Técnicas de Cultivo de Embriones , Desarrollo Embrionario/efectos de los fármacos , Glicoproteínas/farmacología , Péptidos y Proteínas de Señalización Intercelular/farmacología , Porcinos , Trofoblastos/efectos de los fármacos , Anfirregulina , Animales , Núcleo Celular/metabolismo , Medios de Cultivo , Citoplasma/metabolismo , Fertilización In Vitro , Glicoproteínas/análisis , Glicoproteínas/metabolismo , Péptidos y Proteínas de Señalización Intercelular/análisis , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Reacción en Cadena de la Polimerasa , ARN Mensajero/metabolismo , Trofoblastos/metabolismoRESUMEN
To design artificial restriction enzymes, synthetic catalytic centers that effectively hydrolyze linear double-stranded polydeoxyribonucleotides are needed. The Co(III) complex of cyclen (CoCyc) attached to polystyrene derivatives hydrolyzes linearized pUC18 DNA with half-lives as short as 30 min at 25 degrees C. The catalytic activity of CoCyc is enhanced by >150 times on attachment to the resin.
Asunto(s)
ADN/química , ADN/metabolismo , Compuestos Organometálicos/química , Compuestos Organometálicos/metabolismo , Poliestirenos/química , Catálisis , Cobalto/química , HidrólisisRESUMEN
Reactivity of the Co(III) complex of cyclen (CoCyc) in the hydrolytic cleavage of supercoiled pUC18 DNA leading to the formation of the corresponding open circular form was enhanced by >200 times upon attachment of CoCyc to cross-linked polystyrenes. Thus, half-lives as short as 40 min were achieved by the resin-based CoCyc in cleavage of the supercoiled DNA at 4 degrees C.