CSIRO Publishing blank image blank image blank image blank imageBooksblank image blank image blank image blank imageJournalsblank image blank image blank image blank imageAbout Usblank image blank image blank image blank imageShopping Cartblank image blank image blank image You are here: Journals > Reproduction, Fertility and Development   
Reproduction, Fertility and Development
  Vertebrate reproductive science and technology
blank image Search
blank image blank image
blank image
  Advanced Search

Journal Home
About the Journal
Editorial Structure
Online Early
Current Issue
Just Accepted
All Issues
Special Issues
Research Fronts
Virtual Issues
Sample Issue
For Authors
General Information
Submit Article
Author Instructions
Open Access
Awards and Prizes
For Referees
Referee Guidelines
Review an Article
Annual Referee Index
For Subscribers
Subscription Prices
Customer Service
Print Publication Dates
Library Recommendation

blue arrow e-Alerts
blank image
Subscribe to our email Early Alert or RSS feeds for the latest journal papers.

red arrow Submit Article
blank image
Use the online submission system to send us your paper.

red arrow Connect with us
blank image
facebook twitter logo LinkedIn

red arrow Connect with SRB
blank image
facebook TwitterIcon

Affiliated Societies

RFD is the official journal of the International Embryo Transfer Society and the Society for Reproductive Biology.

Article << Previous     |     Next >>   Contents Vol 22(1)


F. Q. Costa A, M. M. Pereira B, S. Wohlres-Viana B, R. V. Serapiao C, B. C. Carvalho D, J. H. M. Viana C, L. A. G. Nogueira A and L. S.A. Camargo C

A Universidade Federal Fluminense, Niterói, Rio de Janeiro, Brazil;
B Universidade Federal de Juiz de Fora, Juiz de Fora, Minas Gerais, Brazil;
C Embrapa Gado de Leite, Juiz de Fora, Minas Gerais, Brazil;
D Epamig, Juiz de Fora, Minas Gerais, Brazil

Export Citation


Heat stress has been shown to have detrimental effects (41°C) during the first 12 h of in vitro maturation on bovine embryo development (Edwards JL and Hansen PJ 1996 Biol. Reprod. 55, 341-346). However, little is known about the effect on gene expression of in vitro-fertilized andparthenogenetic bovine embryos. This study evaluated the gene expression of in vitro-fertilized andparthenogenetic blastocysts derived fromheat-stressed oocytes. The transcripts evaluated were associated with genes encoding proteins involved in blastocoel formation [aquaporin (Aqp) 3 and Na+/K+-ATPase alpha 1; Watson AJ and Barcroft LC 2001 Frontiers Biosci. 6:d708-730] and cell viability (Bax and Peroxiredoxin 1; Van Delft MF and Huang DCS 2006 Cell Res. 16, 203-213; RaguS etal. 2007 PNAS104, 9747-9752). Oocytes were in vitro matured for 12 h at 41°C followed by 12 h at 38.5°C (heat-stressed oocytes; HS) or for 24 h at 38.5°C (non-heat-stressed oocytes; NHS) under 5% CO2. Heat-stressed and NHS oocytes were in vitro fertilized with Holstein sperm (HS-IVF and NHS-IVF subgroups, respectively) or activated with ionomycin and 6-DMAP (HS-PART and NHS-PART subgroups, respectively). Presumptive zygotes were cultured in CR2aa medium under 5% CO2, 5% O2, and 90% N2 at 38.5°C. Embryos at blastocyst stage with same quality grade for all subgroups were obtained from 3 different replicates and distributed in pools of 10 embryos for relative quantification of the target transcripts. RNA extraction and reverse transcription were performed and cDNA quantified by real-time PCR. Transcripts of H2a gene were used as endogenous control, and statistical analysis was performed by pair-wise, fixed reallocation randomization test. Gene expression comparisons were performed between HS-IVF and NHS-IVF, HS-PART and NHS-PART, NHS-PART and NHS-IVF, and HS-PART and HS-IVF subgroups. Blastocyst rate is shown as mean ± SEM and relative expression as n-fold. The heat stress on oocytes during in vitro maturation decreased (P < 0.05, ANOVA) the development of presumptive zygotes to blastocyst stage at Day 8 for in vitro-fertilized (19.9 ± 2.9% and 10.5 ± 2.0% for NHS-IVF and HS-IVF, respectively) and parthenogenetic (33.0 ± 1.8% and 22.8 ± 2.8% for NHS-PART and HS-PART, respectively) embryos. Embryos from the HS-IVF subgroup showed less (P < 0.01) expression of Na+/K+-ATPase alpha 1 (0.67-fold) than did NHS-IVF embryos, whereas no difference was found for others genes. Embryos from the HS-PART subgroup showed less (P < 0.01) expression of Aqp 3 (0.77-fold) and greater (P < 0.05) expression of Bax (1.40-fold) than did NHS-PART embryos. Expression of Aqp 3 was up-regulated (P < 0.01) in NHS-PART (1.42-fold) embryos when compared with NHS-IVF ones, whereas expression of Na+/K+-ATPase alpha 1 (1.42-fold), Bax (1.67-fold), and Peroxiredoxin 1 (1.40-fold) were up-regulated (P < 0.05) in HS-PART embryos when compared with HS-IVF embryos. In conclusion, heat stress on oocytes during in vitro maturation can affect the amount of transcripts of in vitro-fertilized andparthenogenetic blastocysts, suggesting a residual effect on gene expression of bovine embryos.

Financial support was provided by CNPq and FAPEMIG.

Reproduction, Fertility and Development 22(5305) 276–276   http://dx.doi.org/10.1071/RDv22n1Ab236
Published online: 08 December 2009

Top  Email this page

Legal & Privacy | Contact Us | Help


© CSIRO 1996-2016