PMID- 20477985
OWN - NLM
STAT- MEDLINE
DCOM- 20110428
LR  - 20110119
IS  - 1439-0531 (Electronic)
IS  - 0936-6768 (Linking)
VI  - 46
IP  - 1
DP  - 2011 Feb
TI  - Assessment of actin cytoskeleton and nuclei in bovine blastocysts developed under 
      different culture conditions using a novel computer program.
PG  - e46-53
LID - 10.1111/j.1439-0531.2010.01627.x [doi]
AB  - This study was performed to investigate the effects, in terms of nuclear material 
      and actin cytoskeleton quantities (fluorescent pixel counts), of four different 
      bovine blastocyst culturing techniques (in vitro, stepwise in vitro-to-in vivo, 
      or purely in vivo). Cumulus oocyte complexes from abattoir-sourced ovaries were 
      matured in vitro and allocated to four groups: IVP-group embryos developed up to 
      blastocyst stage in vitro. Gamete intra-fallopian transfer (GIFT)-group oocytes 
      were co-incubated with semen for 4 h before transfer to oviducts of heifers. 
      Following in vitro fertilization, cleaved embryos (day 2 of embryo development, 
      day 2-7 group) were transferred into oviducts on day 2. Multiple ovulation embryo 
      transfer (MOET)-group embryos were obtained by superovulating and inseminating 
      heifers; the heifers' genital tracts were flushed at day 7 of blastocyst 
      development. Within each group, ten blastocysts were selected to be 
      differentially dyed (for nuclei and actin cytoskeleton) with fluorescent stains. 
      A novel computer program (ColorAnalyzer) provided differential pixel counts 
      representing organelle quantities. Blastocysts developed only in vivo (MOET 
      group) showed significantly more nuclear material than did blastocysts produced 
      by any other technique. In terms of actin cytoskeleton quantity, blastocysts 
      produced by IVP and by day 2-7 transfer did not differ significantly from each 
      other. Gamete intra-fallopian transfer- and MOET-group embryos showed 
      significantly larger quantities of actin cytoskeleton when compared with any 
      other group and differed significantly from each other. The results of this study 
      indicate that culturing under in vitro conditions, even with part time in vivo 
      techniques, may adversely affect the quantity of blastocyst nuclear material and 
      actin cytoskeleton. The software employed may be useful for culture environment 
      evaluation/developmental competence assessment.
CI  - (c) 2010 Blackwell Verlag GmbH.
FAU - Kuzmany, A
AU  - Kuzmany A
AD  - Reproduction Centre Wieselburg, University of Veterinary Medicine, Vienna, 
      Austria. anna.kuzmany@boku.ac.at
FAU - Havlicek, V
AU  - Havlicek V
FAU - Brem, G
AU  - Brem G
FAU - Walter, I
AU  - Walter I
FAU - Besenfelder, U
AU  - Besenfelder U
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PL  - Germany
TA  - Reprod Domest Anim
JT  - Reproduction in domestic animals = Zuchthygiene
JID - 9015668
RN  - 0 (Actins)
SB  - IM
MH  - Actins/*analysis
MH  - Animals
MH  - Blastocyst/physiology/*ultrastructure
MH  - Cattle/*embryology
MH  - Cell Nucleus/*ultrastructure
MH  - Cytoskeleton/chemistry/*ultrastructure
MH  - Embryo Culture Techniques/methods/*veterinary
MH  - Embryo Transfer/methods/veterinary
MH  - Female
MH  - Fertilization in Vitro/veterinary
MH  - Gamete Intrafallopian Transfer/veterinary
MH  - Insemination, Artificial/veterinary
MH  - Male
MH  - Software
MH  - Tissue and Organ Harvesting/methods/veterinary
EDAT- 2010/05/19 06:00
MHDA- 2011/04/29 06:00
CRDT- 2010/05/19 06:00
PHST- 2010/05/19 06:00 [entrez]
PHST- 2010/05/19 06:00 [pubmed]
PHST- 2011/04/29 06:00 [medline]
AID - RDA1627 [pii]
AID - 10.1111/j.1439-0531.2010.01627.x [doi]
PST - ppublish
SO  - Reprod Domest Anim. 2011 Feb;46(1):e46-53. doi: 10.1111/j.1439-0531.2010.01627.x.