Analysis of in vivo- and in vitro-derived pig expanded blastocysts based on DNA fragmentation

• Autorzy: Bryla M , Trzcinska M. , Wieczorek J.

Warianty tytułu

PL

Ocena fragmentacji jadrowego DNA w zarodkach swini w stadium ekspandujacej blastocysty uzyskanych in vivo i in vitro

• Języki publikacji: EN

Abstrakty

EN

The aim of the study was to investigate the competence of in vivo- and in vitro-derived pig expanded blastocysts by analysing DNA fragmentation using TUNEL. A total of 533 porcine expanded blastocysts were examined, and results were evaluated using Fisher’s test. Significant differences in the incidence of fragmented nuclei (detected by the TUNEL reaction) and all nuclei (detected by DAPI) were identified between in vivo- and in vitro-derived embryos at the expanded blastocyst stage. The total numbers of nuclei observed in in vivo-derived embryos were significantly different from those in in vitro-cultured embryos (89.1±13.4 and 47.7±25.1, respectively). TUNEL index in In vitro-cultured embryos (28.3%) was significantly higher (P<0.01) than in in vivo-derived blastocysts (4%). These findings indicate that in vivo- and in vitro-derived expanded blastocysts consisting of a small number of cells are characterized by a high incidence of DNA fragmentation. The total number of nuclei in in vivo- and in vitro-derived blastocysts correlated negatively with the number of TUNEL-positive nuclei (r = -0.51; P<0.0001) and the TUNEL index (r = -0.69; P<0.0001), whereas the number of TUNEL-positive nuclei was positively correlated with the TUNEL index (r = 0.95;P<0.0001). Moreover, significant differences were observed between embryos collected from individual experiments.

PL

Celem pracy była ocena jakości zarodków świni uzyskanych in vivo i hodowanych in vitro,ocenianych na podstawie fragmentacji DNA z zastosowaniem metody TUNEL i całkowitej liczby jąder (wyznakowanych barwnikiem DAPI). Badania przeprowadzono na 533 zarodkach w stadium ekspandującej blastocysty, a uzyskane wyniki szacowano testem Fishera. Istotnie wyższą całkowitą liczbę jąder i wielkość indeksu apoptotycznego zaobserwowano w zarodkach poddanych hodowli In vitro (odpowiednio 47,7±25,11 i 28,3%) w porównaniu z zarodkami uzyskanymi in vivo (odpowiednio 89,1±13,45 i 4%). Uzyskane wyniki wskazują też, że niskiej całkowitej liczbie jąder towarzyszy nasilenie fragmentacji DNA zarówno w grupie zarodków uzyskanych in vivo jak i in vitro. Analiza zarodków wykazała ponadto ujemną korelację między całkowitą liczbą jąder a liczbą jąder apoptotycznych (r =-0,51, P<0,0001) i indeksem apoptotycznym (r = -0,69, P<0,0001). Stwierdzono dodatnią korelację między liczbą jąder apoptotycznych a indeksem apoptotycznym (r = 0,95, P<0,0001). Dodatkowo wykazano istotne różnice między jakością zarodków (in vivo i in vitro) uzyskanych w poszczególnych doświadczeniach.

Słowa kluczowe

EN

in vitro; DNA fragmentation; blastocyst; pig; TUNEL method; porcine expanded blastocyst; in vivo

• Wydawca: -
• Czasopismo: Animal Science Papers and Reports
• Rocznik: 2009
• Tom: 27
• Numer: 1
• Opis fizyczny: p.59-68,ref.

Twórcy

autor

Bryla M

• National Research Institute of Animal Production, Krakowska 1, 32-083 Balice, Poland

autor

Trzcinska M.

autor

Wieczorek J.

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