Type

Journal Article

Authors

A Gutiérrez-Adán
J de la Fuente
B Pintado
R Arroyo-García
M P Boland
P. Lonergan
D Rizos

Subjects

Biochemistry

Topics
gene expression superoxide dismutase lymphokines receptors osm lif lif protein human reverse transcriptase polymerase chain reaction gjb3 protein human female proto oncogene proteins cattle chemistry superovulation genetics cryopreservation receptors cytokine sarcosine oxidase culture techniques leukemia inhibitory factor oxidoreductases n demethylating coculture techniques growth inhibitors physiology sheep bcl 2 associated x protein lifr protein human connexin 43 interleukin 6 polymorphism genetic proto oncogene proteins c bcl 2 animals leukemia inhibitory factor receptor alpha subunit fertilization in vitro blastocyst fallopian tubes veterinary embryology insemination artificial connexins rna messenger

Analysis of differential messenger RNA expression between bovine blastocysts produced in different culture systems: implications for blastocyst quality. (2002)

Abstract Using reverse transcriptase-amplified fragment length polymorphism (RT-AFLP) analysis of differential mRNA expression and semiquantitative reverse transcriptase-polymerase chain reaction, we compared mRNA expression in bovine blastocysts from 4 sources, known to differ in quality in terms of their ability to withstand cryopreservation: 1) in vitro culture in synthetic oviduct fluid of in vitro-matured (IVM)/in vitro fertilized (IVF) zygotes; 2) in vitro culture in TCM-199 supplemented with granulosa cells (coculture) of IVM/IVF zygotes; 3) in vivo culture in the ewe oviduct of IVM/IVF zygotes; or 4) superovulation, artificial insemination, and nonsurgical embryo recovery. Total mRNA was isolated from pools of blastocysts and reverse transcription was performed. Triplicate reactions from each sample were displayed, and only consistent banding variations were recorded. Using AFLP-differential display assay, we found that cDNA banding patterns are highly conserved between the 4 groups of blastocysts studied; however, there was a difference of 7% in bands either missing or expressed across the groups. Fifty bands were reamplified, and a sequence comparison search revealed similarity of 14 isolated fragments to ribosomal and mitochondrial genes, 16 matched to described cDNA, and 20 corresponded to unknown sequences that may represent novel genes. The study of 7 differentially expressed mRNAs known to be involved in developmental process in the embryo suggests roles for apoptosis, oxidative stress, gap junctions, and differentiation in the determination of embryo quality. The aberrant transcription patterns detected in in vitro-produced bovine embryos compared with those produced in vivo may explain their reduced quality in terms of viability after cryopreservation.
Collections Ireland -> University College Dublin -> PubMed

Full list of authors on original publication

A Gutiérrez-Adán, J de la Fuente, B Pintado, R Arroyo-García, M P Boland, P. Lonergan, D Rizos

Experts in our system

1
M P Boland
University College Dublin
Total Publications: 103
 
2
P. Lonergan
University College Dublin
Total Publications: 190
 
3
D Rizos
University College Dublin
Total Publications: 42