Hostname: page-component-848d4c4894-m9kch Total loading time: 0 Render date: 2024-06-01T18:41:43.530Z Has data issue: false hasContentIssue false

Apoptosis in the early bovine embryo

Published online by Cambridge University Press:  01 February 2000

Christie Matwee
Affiliation:
Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada N1G 2WI
Dean H. Betts
Affiliation:
Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada N1G 2WI
W. Allan King
Affiliation:
Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada N1G 2WI

Abstract

Cell death occurs during early development in vivo and in vitro, although little is known about the mechanism of blastomere death and the relation to embryonic loss. Apoptosis, characterised by chromatin condensation, DNA fragmentation and membrane blebbing, occurs without damage to surrounding cells in contrast to necrosis. Bovine oocytes and in vitro fertilised embryos (total n = 449) were analysed for (1) DNA fragmentation using terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) and (2) morphological features of apoptosis. TUNEL labelling was detected in immature and mature oocytes (7%, n = 57 and 23%, n = 60, respectively), and at least one cell of 8- to 16-cell embryos (5%, n = 57), morulae/early blastocysts (79%, n = 39) and expanded/hatched blastocysts (100%, n = 48). In contrast, TUNEL labelling was not detected in zygotes (n = 61), 2-cell embryos (n = 46) or 3- to 7-cell embryos (n = 81). Chromatin condensation, nuclear fragmentation, absence of neighbouring cell destruction and extrusion of cells was frequent among advanced stage embryos. Although not detected during early cleavage under standard conditions, TUNEL labelling indicative of apoptosis was induced by treatment with 10 μM staurosporine for 30 h in 95% of cleavage stage embryos (n = 59). Determination of the expression and localisation of the p53 tumour suppressor gene using reverse transcription polymerase chain reaction and whole-mount immunofluorescence revealed that although p53 transcripts were present throughout early development, nuclear localisation of p53 protein could not be detected in any blastocyst suggesting p53-independent apoptosis. This study has shown that apoptosis is dependent on embryonic developmental stage after standard culture. This suggests that bovine embryos become more capable of accommodating damaged or abnormal cells as development proceeds.

Type
Research Article
Copyright
© 2000 Cambridge University Press

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)