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Expression Pattern of Apoptotic Genes in Vitrified‐Thawed Bovine Oocytes
Author(s) -
Anchamparuthy VM,
Pearson RE,
Gwazdauskas FC
Publication year - 2010
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/j.1439-0531.2009.01527.x
Subject(s) - vitrification , andrology , fas ligand , germinal vesicle , apoptosis , biology , gene expression , embryo , messenger rna , microbiology and biotechnology , oocyte , gene , programmed cell death , biochemistry , medicine
Contents This study describes a method for quantification of transcripts from low numbers of bovine oocytes using real time RT‐PCR. The objective was to evaluate the expression pattern of apoptotic genes ( Fas , FasL , Bax and Bcl‐2 ) in vitrified‐thawed oocytes. Oocytes were evaluated at germinal vesicle stage; at 15 h of maturation; after vitrification and warming at 15 h of maturation and at 9 h of additional maturation. All transcripts showed an increase in at least 1.2‐fold change post‐vitrification warming, but the levels tended to decrease at 9 h of maturation post‐vitrification warming. Transcript abundance for Fas mRNA was 1.4‐fold for oocytes after vitrification and warming. The level of Fas mRNA upon maturation was 0.8‐fold. The increase in the abundance of FasL mRNA was 2.1, while it was 0.5‐fold relative to control. Vitrification resulted in 1.5‐fold change in Bax mRNA expression in oocytes. After 9 h of maturation post‐vitrification warming, the level for Bax mRNA was 0.6‐fold. The mRNA for Bcl‐2 was nearly the same after vitrification and warming. The abundance of mRNA for Bcl‐2 was 1.2‐fold in vitrified oocytes and fell (p = 0.05) to 0.5 at 9 h of maturation post‐vitrification and warming. The up‐regulation of apoptotic genes in vitrified oocytes may be an early indicator of reduced developmental competence following vitrification. Yet, results from terminal deoxynucleotidyl transferase dUTP nick end labelling and caspase assays did not support the evidence of apoptosis in embryos derived from large numbers of vitrified oocytes.